← Product Code OYB · P100042 # APTIMA HPV ASSAY (P100042) _Gen-Probe Incorporated · OYB · Oct 28, 2011 · Microbiology · APPR_ **Canonical URL:** https://fda.innolitics.com/device/P100042 ## Device Facts - **Applicant:** Gen-Probe Incorporated - **Product Code:** OYB - **Decision Date:** Oct 28, 2011 - **Decision:** APPR - **Device Class:** Class 3 - **Review Panel:** Microbiology ## Intended Use The APTIMA HPV Assay is an in vitro nucleic acid amplification test for the qualitative detection of E6/E7 viral messenger RNA (mRNA) from 14 high-risk types of human papillomavirus (HPV) in cervical specimens. The high-risk HPV types detected by the assay include: 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68. The APTIMA HPV Assay does not discriminate between the 14 high-risk types. Cervical specimens in ThinPrep Pap Test vials containing PreservCyt Solution and collected with broom-type or cytobrush/spatula collection devices* may be tested with the APTIMA HPV Assay. The assay is used with the TIGRIS DTS System. The use of the test is indicated: 1. To screen patients 21 years and older with atypical squamous cells of undetermined significance (ASC-US) cervical cytology results to determine the need for referral to colposcopy. The results of this test are not intended to prevent women from proceeding to colposcopy. 2. In women 30 years and older, the APTIMA HPV Assay can be used with cervical cytology to adjunctively screen to assess the presence or absence of high-risk HPV types. This information, together with the physician's assessment of cytology history, other risk factors, and professional guidelines, may be used to guide patient management. ## Device Story The APTIMA HPV Assay is an in vitro diagnostic test for detecting E6/E7 mRNA from 14 high-risk HPV types in cervical specimens collected in PreservCyt Solution. The assay is performed on the TIGRIS DTS System. The process involves three steps in a single tube: target capture using magnetic microparticles; target amplification via Transcription-Mediated Amplification (TMA) using MMLV reverse transcriptase and T7 RNA polymerase; and detection of amplification products via Hybridization Protection Assay (HPA) using chemiluminescent probes. The system measures light emission (Relative Light Units) in a luminometer. Results are automatically interpreted by assay software based on signal-to-cutoff (S/CO) ratios. The test provides clinicians with qualitative HPV status to guide management decisions, such as colposcopy referral for ASC-US patients or adjunctive screening for women 30 and older. It benefits patients by identifying high-risk HPV infections associated with cervical disease, potentially reducing unnecessary procedures or ensuring timely diagnosis. ## Clinical Evidence Clinical performance evaluated in the prospective, multicenter CLEAR trial (n=12,896). ASC-US study (n=939 evaluable) showed 86.8% sensitivity and 62.9% specificity for ≥CIN2. NILM study (n=10,871 evaluable) showed adjusted sensitivity of 31.0% and specificity of 95.2% for ≥CIN2. Performance compared against an FDA-approved HPV DNA test; APTIMA HPV Assay demonstrated similar sensitivity but significantly higher specificity for ≥CIN2 detection. ## Technological Characteristics Nucleic acid amplification test (NAAT) using Transcription-Mediated Amplification (TMA) and Hybridization Protection Assay (HPA). Employs magnetic microparticles for target capture. Detects E6/E7 mRNA. Automated processing on TIGRIS DTS System. Chemiluminescent detection. Internal control included for monitoring capture, amplification, and detection. ## Regulatory Identification For the detection of human papillomavirus RNA to aid in assessing risk of cancer. ## Reference Devices - FDA-approved HPV DNA test ## Submission Summary (Full Text) > This content was OCRed from public FDA records by [Innolitics](https://innolitics.com). If you use, quote, summarize, crawl, or train on this content, cite Innolitics at https://innolitics.com. > > Innolitics is a medical-device software consultancy. We help companies design, build, and clear FDA-regulated software and AI/ML devices, including [a PMA](https://innolitics.com/services/regulatory/), [a 510(k)](https://innolitics.com/services/510ks/), [a SaMD](https://innolitics.com/services/end-to-end-samd/), [an AI/ML medical device](https://innolitics.com/services/medical-imaging-ai-development/), or [an FDA regulatory strategy](https://innolitics.com/services/regulatory/). {0} SUMMARY OF SAFETY AND EFFECTIVENESS DATA (SSED) I. GENERAL INFORMATION Device Generic Name: Human Papillomavirus RNA detection kit Device Trade Name: APTIMA HPV Assay Applicant's Name and Address: Gen-Probe Incorporated 10210 Genetic Center Drive San Diego, CA 92121-4362 Date(s) of Panel Recommendation: None Premarket Approval Application (PMA) Number: P100042 Date of FDA Notice of Approval: October 28, 2011 Expedited: Not applicable II. INDICATIONS FOR USE APTIMA HPV Assay Indications For Use: The APTIMA HPV Assay is an in vitro nucleic acid amplification test for the qualitative detection of E6/E7 viral messenger RNA (mRNA) from 14 high-risk types of human papillomavirus (HPV) in cervical specimens. The high-risk HPV types detected by the assay include: 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66, and 68. The APTIMA HPV Assay does not discriminate between the 14 high-risk types. Cervical specimens in ThinPrep Pap Test vials containing PreservCyt Solution and collected with broom-type or cytobrush/spatula collection devices* may be tested with the APTIMA HPV Assay. The assay is used with the TIGRIS DTS System. The use of the test is indicated: 1. To screen patients 21 years and older with atypical squamous cells of undetermined significance (ASC-US) cervical cytology results to determine the need for referral to colposcopy. The results of this test are not intended to prevent women from proceeding to colposcopy. 2. In women 30 years and older, the APTIMA HPV Assay can be used with cervical cytology to adjunctively screen to assess the presence or absence of high-risk HPV types. This information, together with the physician's assessment of cytology history, other risk factors, and professional guidelines, may be used to guide patient management. PMA P100042: FDA Summary of Safety and Effectiveness Data {1} * Broom-type device (e.g., Wallach Pipette) or endocervical brush/spatula. ## III. CONTRAINDICATIONS None. ## IV. WARNINGS AND PRECAUTIONS The warnings and precautions can be found in the APTIMA HPV Assay labeling. ## V. DEVICE DESCRIPTION The APTIMA HPV Assay involves three main steps, which take place in a single tube: target capture; target amplification by Transcription-Mediated Amplification (TMA); and detection of the amplification products (amplicon) by the Hybridization Protection Assay (HPA). The assay incorporates an internal control to monitor nucleic acid capture, amplification, and detection, as well as operator or instrument error. Specimens are transferred to a tube containing specimen transport media (STM) that lyses the cells, releases the mRNA, and protects it from degradation during storage. When the APTIMA HPV Assay is performed, the target mRNA is isolated from the specimen by use of capture oligomers that are linked to magnetic microparticles. The capture oligomers contain sequences complementary to specific regions of the HPV mRNA target molecules as well as a string of deoxyadenosine residues. During the hybridization step, the sequence-specific regions of the capture oligomers bind to specific regions of the HPV mRNA target molecule. The capture oligomer-target complex is then captured out of solution by decreasing the temperature of the reaction to room temperature. This temperature reduction allows hybridization to occur between the deoxyadenosine region on the capture oligomer and the poly-deoxythymidine molecules that are covalently attached to the magnetic particles. The microparticles, including the captured HPV mRNA target molecules bound to them, are pulled to the side of the reaction tube using magnets and the supernatant is aspirated. The particles are washed to remove residual specimen matrix that may contain amplification inhibitors. After target capture is complete, the HPV mRNA is amplified using TMA, which is a transcription-based nucleic acid amplification method that utilizes two enzymes, MMLV reverse transcriptase and T7 RNA polymerase. The reverse transcriptase is used to generate a DNA copy of the target mRNA sequence containing a promoter sequence for T7 RNA polymerase. T7 RNA polymerase produces multiple copies of RNA amplicon from the DNA copy template. Detection of the amplicon is achieved by HPA using single-stranded nucleic acid probes with chemiluminescent labels that are complementary to the amplicon. The labeled nucleic acid probes hybridize specifically to the amplicon. The Selection Reagent differentiates between hybridized and unhybridized probes by inactivating the label on PMA P100042: FDA Summary of Safety and Effectiveness Data {2} the unhybridized probes. During the detection step, light emitted from the labeled RNA-DNA hybrids is measured as photon signals called Relative Light Units (RLU) in a luminometer. Final assay results are interpreted based on the analyte signal-to-cutoff (S/CO). Internal Control is added to each reaction via the Target Capture Reagent. The Internal Control monitors the target capture, amplification, and detection steps of the assay. Internal Control signal in each reaction is discriminated from the HPV signal by the differential kinetics of light emission from probes with different labels. Internal Control-specific amplicon is detected using a probe with a rapid emission of light (flasher). Amplicon specific to HPV is detected using probes with relatively slower kinetics of light emission (glower). The Dual Kinetic Assay (DKA) is the method used to differentiate between the signals from the flasher and glower labels. Additional details can be found in the operator’s manual for the device. ## Test Interpretation Assay test results are automatically determined by the assay software. A test result may be negative, positive, or invalid as determined by the Internal Control (IC) RLU and the signal-to-cutoff (S/CO) for the Analyte. A test result may also be invalid due to other parameters (abnormal kinetic curve shape) being outside the normal expected ranges. Invalid test results should be repeated. | APTIMA HPV Assay Result | Criteria | | --- | --- | | Negative | Analyte S/CO < 0.50 Internal Control ≥ IC Cutoff Internal Control ≤ 2,000,000 RLU | | Positive | Analyte S/CO ≥ 0.50 Internal Control ≤ 2,000,000 RLU Analyte ≤ 13,000,000 RLU | | Invalid | Analyte S/CO < 0.50 and Internal Control < IC Cutoff Or Internal Control > 2,000,000 RLU Or Analyte > 13,000,000 RLU | NOTE: Negative results are not intended to prevent women from proceeding to colposcopy. NOTE: Negative results indicate HPV E6/E7 mRNA was not detected. NOTE: Negative results may occur with HPV E6/E7 mRNA concentrations that are below the pre-set threshold. NOTE: Positive results indicate the presence of HPV E6/E7 mRNA of any one or more of the high risk types. PMA P100042: FDA Summary of Safety and Effectiveness Data {3} NOTE: Results of this test should only be interpreted in conjunction with information available from clinical evaluation of the patient and patient history. ## VI. ALTERNATIVE PRACTICES AND PROCEDURES The patient’s age, medical history and thorough physical examination, including cytology, will provide further information on a patient’s risk of cervical disease, as well as the need for referral to colposcopy. The APTIMA HPV Assay should only be used in conjunction with this clinical information in accordance with appropriate patient management procedures. Three alternatives for the detection of high-risk HPV are currently approved in the United States. All three alternative devices detect HPV DNA instead HPV mRNA. Each HPV detection method has its own advantages and disadvantages. A patient should fully discuss these alternatives with her physician to select the screening method(s) that best meets expectations and her lifestyle. ## VII. MARKETING HISTORY The APTIMA HPV Assay is CE marked for use in the EU and is approved for use in multiple countries. A list of all countries where the APTIMA HPV Assay is marketed as of September 2011 is provided below. The APTIMA HPV Assay has not been withdrawn from these markets for any reason. - Australia - Austria - Bahamas - Belgium - Bermuda - Cambodia - Chile - Colombia - Czech Republic - Denmark - Dominican Republic - Finland - France - Germany - Greece - Honduras - Hong Kong - Hungary - Indonesia - Ireland - Israel - Italy - Kenya - Latvia - Libya - Luxembourg - Madagascar - Malaysia - Mali - Namibia - Netherlands - Pakistan - Peru - Philippines - Poland - Portugal - Saudi Arabia - Slovakia - Slovenia - South Africa - Spain - Sweden - Tunisia - UAE - UK - Vietnam ## VIII. POTENTIAL ADVERSE EFFECTS OF THE DEVICE ON HEALTH Below is a list of the potential adverse effects (e.g., complications) associated with the use of the device. As with any in vitro diagnostic test, the potential adverse effects are PMA P100042: FDA Summary of Safety and Effectiveness Data page 4 {4} associated with incorrect test results or result interpretations. Failure of this device to perform as expected or failure to correctly interpret results may lead to incorrect HPV test results and subsequently, improper patient management decisions in cervical cancer screening and treatment. False negative results may lead to delays in the timely diagnosis of cervical cancer and treatment, allowing an undetected condition to worsen and potentially increasing morbidity and mortality. False positive results could lead many women to unnecessarily undergo more frequent screening and potentially invasive procedures such as colposcopy and biopsy. ## IX. SUMMARY OF PRECLINICAL STUDIES ### A. Laboratory Studies #### Clinical Cutoff Determination for the APTIMA HPV Assay The clinical cutoff for detecting high-grade cervical disease (≥CIN2) for the APTIMA HPV Assay was established based on the evaluation of approximately 1000 women with ASC-US cytology results enrolled into the ASC-US Study. The method used for selection of the cutoff was chosen to achieve the maximum sensitivity for detecting ≥CIN2 while maintaining a clinically acceptable level of specificity in the ASC-US population. Based on the method described above, the cutoff for the APTIMA HPV Assay was set at 0.50 S/CO. #### Limit of Detection at the Clinical Cutoff The Limit of Detection (LOD) at the clinical cutoff is the concentration of HPV RNA that gives a positive result (above the clinical cutoff) 95% of the time. The LoD of the APTIMA HPV Assay was determined by testing individual negative clinical PreservCyt liquid cytology specimens spiked with HPV in vitro transcripts or infected cells at various concentrations. Thirty replicates of each copy level were tested with each of three reagent lots for a total of 90 replicates. Testing was performed over 14 days, with 1 to 14 runs performed per day and 30 replicates of a given genotype tested in each run. The 95% detection limit was calculated from Probit regression analysis of the positivity results for each dilutional panel. The Probit analysis results, in the table below, show that HPV 16, 18, 31, 33, 35, 39, 45, 58, 59, 66, and 68 had 95% detection limits less than 100 copies/reaction; and types 51, 52, and 56 had 95% detection limits between 100 and 300 copies/reaction. The four cell lines tested had 95% detection limits less than 1 cell/reaction. Each target analyte was diluted in STM prior to adding PreservCyt media at a 1:2.9 ratio for these LOD panel specimens. Therefore, the results of this study do not reflect any effects of sampling from a heterogenous non-lysed whole cell suspension prior to PreservCyt specimen transfer to STM. Limit of Detection at Clinical Cutoff of the APTIMA HPV Assay | Target | Limit of Detection* (95% CI) | | --- | --- | | HPV 16 | 27.1 (21.7 - 36.1) | | HPV 18 | 54.1 (42.9 - 73.2) | | HPV 31 | 11.5 (9.2 - 15.6) | PMA P100042: FDA Summary of Safety and Effectiveness Data {5} | Target | Limit of Detection* (95% CI) | | --- | --- | | HPV 33 | 34.4 (26.9 - 47.4) | | HPV 35 | 31.9 (23.6 - 47.3) | | HPV 39 | 20.5 (16.2 - 28.0) | | HPV 45 | 26.3 (21.1 - 34.9) | | HPV 51 | 132.9 (116.7 - 159.6) | | HPV 52 | 240.2 (191.4 - 320.3) | | HPV 56 | 105.5 (88.0 - 133.2) | | HPV 58 | 71.4 (55.9 - 97.3) | | HPV 59 | 62.3 (47.7 - 89.7) | | HPV 66 | 208.0 (168.4 - 270.0) | | HPV 68 | 47.1 (35.9 - 67.9) | | SiHa | 0.288 (0.223 - 0.401) | | HeLa | 0.029 (0.024 - 0.036) | | ME180 | 0.0012 (0.0009 - 0.0018) | | MS751 | 0.018 (0.015 - 0.024) | * Copies per reaction for in vitro transcripts and cells per reaction for cell lines ## Assay Precision APTIMA HPV Assay precision was evaluated in two studies using the same 20-member panel. Study 1 was conducted at 3 external testing sites to determine assay reproducibility. Study 2 was conducted in-house to measure assay repeatability. The panel included 10 HPV-positive members with concentrations at or above the limit of detection of the assay (expected positivity: ≥ 95%), 4 HPV-positive members with concentrations below the limit of detection of the assay (expected positivity: >0% to <25%), and 6 HPV-negative members. HPV-positive panel members were prepared by spiking in vitro RNA transcripts (IVT) into specimen transport medium (STM) or HPV-infected cultured cells (SiHa, HeLa, ME180 and MS751; ATCC, Manassas, Virginia) into PreservCyt Solution. HPV-negative panel members were prepared with STM or pooled residual PreservCyt Solution specimens. In Study 1, 2 operators at each of the 3 testing sites (1 instrument per site) performed 1 APTIMA HPV Assay worklist per day over 3 days for each of 3 reagent lots. Each worklist contained 3 replicates of each of the reproducibility panel members. One hundred sixty-two (162) individual sample tubes were tested for each panel member (3 sites x 1 instrument x 2 operators x 3 lots x 3 worklists x 3 replicates). In Study 2, testing was conducted in-house over 20 days with a total of 162 reactions tested for each panel member (1 site x 3 instruments x 3 operators x 3 lots x 2 worklists x 3 replicates). The panel members are described in the tables below, along with a summary of the agreement with expected results and analyte S/CO values at the 2.5th, 50th and 97.5th percentiles of the S/CO distribution. PMA P100042: FDA Summary of Safety and Effectiveness Data {6} APTIMA HPV Assay Reproducibility Study 1 and 2: Panel Description, Positive Agreement, and Percentile Distribution of Analyte S/CO Values for Panel Members with Expected Positive Results | Panel Description (copies or cells/reaction) | Study 1 (3 testing sites) | | | | Study 2 (1 testing site) | | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | % positive agreement (95% CI) | Analyte S/CO Percentile | | | % positive agreement (95% CI) | Analyte S/CO Percentile | | | | | | 2.5^{th} | 50^{th} | 97.5^{th} | | 2.5^{th} | 50^{th} | 97.5^{th} | | HPV 16 & HPV 18 IVT (100 copies) | 100 (161/161) (97.7, 100) | 20.7 | 23.5 | 26.3 | 100 (162/162) (97.7, 100) | 20.1 | 23.2 | 26.3 | | SiHa cells (3 cells) & HeLa cells (7.5 cells) | 100 (162/162) (97.7, 100) | 11.0 | 15.3 | 28.0 | 100 (162/162) (97.7, 100) | 12.5 | 16.5 | 28.0 | | HPV 18 IVT (100 copies) | 100 (162/162) (97.7, 100) | 8.5 | 11.8 | 13.9 | 100 (160/160) (97.7, 100) | 9.0 | 11.9 | 14.7 | | HPV 16 IVT (100 copies) | 100 (162/162) (97.7, 100) | 9.9 | 10.8 | 11.6 | 100 (162/162) (97.7, 100) | 9.3 | 10.9 | 11.7 | | MS751 cells (1 cell) | 99.4 (161/162) (96.6, 99.9) | 6.1 | 13.9 | 16.0 | 96.9 (157/162) (93.0, 98.7) | 0 | 14.5 | 16.6 | | ME180 cells (0.3 cells) | 95.1 (154/162) (90.6, 97.5) | 0 | 7.3 | 9.3 | 93.2 (151/162) (88.3, 96.2) | 0 | 6.5 | 9.4 | | HPV 18 IVT (30 copies) | 99.4 (161/162) (96.6, 99.9) | 3.0 | 9.3 | 13.3 | 100 (162/162) (97.7, 100) | 4.3 | 8.7 | 13.6 | | HPV 16 IVT (30 copies) | 100 (162/162) (97.7, 100) | 8.2 | 10.9 | 11.8 | 97.5 (158/162) (93.8, 99.0) | 4.3 | 11.0 | 11.9 | | HeLa cells (2.5 cells) | 100 (162/162) (97.7, 100) | 6.6 | 12.9 | 15.8 | 95.6 (152/159) (91.2, 97.9) | 0 | 12.9 | 16.5 | | SiHa cells (1 cell)* | 77.2 (125/162) (70.1, 83.0) | 0 | 10.5 | 12.1 | 79.0 (128/162) (72.1, 84.6) | 0 | 10.5 | 11.8 | IVT = in vitro transcript. IVT was spiked into STM and cells were spiked into PreservCyt Solution. *Expected % positive agreement ~95%; observed lower possibly due to manufacturing variability of the panel member. PMA P100042: FDA Summary of Safety and Effectiveness Data {7} APTIMA HPV Assay Reproducibility Study 1 and 2: Panel Description, Negative Agreement, and Percentile Distribution of Analyte S/CO Values for Panel Members with Expected Negative Results | Panel Description (copies or cells/reaction) | Study 1 (3 testing sites) | | | | Study 2 (1 testing site) | | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | % negative agreement (95% CI) | Analyte S/CO Percentile | | | % negative agreement (95% CI) | Analyte S/CO Percentile | | | | | | 2.5^{th} | 50^{th} | 97.5^{th} | | 2.5^{th} | 50^{th} | 97.5^{th} | | HPV 18 IVT (1 copy)* | 78.8 (126/160) (71.8, 84.4) | 0 | 0 | 4.6 | 83.3 (135/162) (76.8, 88.3) | 0 | 0 | 5.2 | | HPV 16 IVT (1 copy)* | 80.9 (131/162) (74.1, 86.2) | 0 | 0 | 10.7 | 88.3 (143/162) (82.4, 92.4) | 0 | 0 | 11.3 | | HeLa cells (0.05 cells)* | 79.0 (128/162) (72.1, 84.6) | 0 | 0 | 10.4 | 82.1 (133/162) (75.5, 87.2) | 0 | 0 | 11.6 | | SiHa cells (0.03 cells)* | 93.8 (152/162) (89.0, 96.6) | 0 | 0 | 10.7 | 95.7 (155/162) (91.4, 97.9) | 0 | 0 | 6.9 | | STM Lot 1 | 100 (162/162) (97.7, 100) | 0 | 0 | 0.1 | 100 (162/162) (97.7, 100) | 0 | 0 | 0 | | STM Lot 2 | 99.4 (160/161) (96.6, 99.9) | 0 | 0 | 0.2 | 100 (162/162) (97.7, 100) | 0 | 0 | 0 | | STM Lot 3 | 99.4 (161/162) (96.6, 99.9) | 0 | 0 | 0.1 | 99.4 (161/162) (96.6, 99.9) | 0 | 0 | 0 | | ThinPrep Pool 1 | 97.5 (158/162) (93.8, 99.0) | 0 | 0 | 0.3 | 97.5 (158/162) (93.8, 99.0) | 0 | 0 | 0.3 | | ThinPrep Pool 2 | 96.9 (157/162) (93.0, 98.7) | 0 | 0 | 0.7 | 96.3 (156/162) (92.2, 98.3) | 0 | 0 | 1.6 | | ThinPrep Pool 3 | 100 (162/162) (97.7, 100) | 0 | 0 | 0.2 | 99.4 (161/162) (96.6, 99.9) | 0 | 0 | 0 | STM = specimen transport medium; IVT = in vitro transcript. IVT was spiked into STM and cells were spiked into PreservCyt Solution. * Expected % negative agreement > 75% and < 100%. The analyte S/CO variability for the panel members with expected positive results is shown in the tables below for Study 1 and Study 2. Positive agreement for the HPV-positive panel members with concentrations at or above the limit of detection of the assay ranged from 95.1% to 100% in Study 1 and from 93.2% to 100% in Study 2 for 9 of the 10 panel members. The remaining HPV-positive panel member yielded 77.2% agreement in Study 1 and 79.0% agreement in Study 2, which was lower than expected, but was consistent between the 2 studies. Negative agreement for the HPV-high negative panel members with concentrations below the limit of detection of the assay ranged from 78.8% to 93.8% in Study 1 and from 82.1% to 95.7% in Study 2. Agreement with expected results for the HPV-negative panel members ranged from 96.9% to 100% in Study 1 and from 96.3% to 100% in Study 2. PMA P100042: FDA Summary of Safety and Effectiveness Data page 8 {8} APTIMA HPV Assay Reproducibility Study 1: Signal Variability for Panel Members With Expected Positive Results | Panel Description (copies or cells/reaction) | n | Mean S/CO | Between Sites | | Between Operators | | Between Lots | | Between Worklists | | Within Worklists | | Total | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | | | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | | HPV 16 & HPV 18 IVT (100 copies) | 161^ | 23.4 | 0.1 | 0.4 | 0.1 | 0.4 | 0.9 | 4.0 | 0 | 0 | 1.6 | 7.0 | 1.9 | 8.1 | | SiHa cells (3 cells) & HeLa cells (7.5 cells) | 162 | 17.9 | 0 | 0 | 1.4 | 8.1 | 0 | 0 | 0.6 | 3.1 | 5.1 | 28.6 | 5.3 | 29.9 | | HPV 18 IVT (100 copies) | 162 | 11.8 | 0 | 0 | 0 | 0 | 0.8 | 6.4 | 0.1 | 0.9 | 1.2 | 10.1 | 1.4 | 12.0 | | HPV 16 IVT (100 copies) | 162 | 10.8 | 0.2 | 1.5 | 0 | 0 | 0.1 | 1.1 | 0.3 | 2.6 | 0.3 | 3.1 | 0.5 | 4.5 | | MS751 cells (1 cell) | 162 | 13.3 | 0.3 | 2.1 | 0 | 0 | 1.0 | 7.8 | 0.9 | 7.1 | 2.2 | 16.2 | 2.6 | 19.4 | | ME180 cells (0.3 cells) | 162 | 6.5 | 0.2 | 3.2 | 0 | 0 | 0.6 | 8.6 | 0.4 | 5.5 | 2.4 | 36.2 | 2.5 | 37.7 | | HPV 18 IVT (30 copies) | 162 | 9.0 | 0.7 | 7.3 | 0 | 0 | 0.7 | 7.2 | 0.8 | 8.3 | 2.3 | 25.3 | 2.6 | 28.5 | | HPV 16 IVT (30 copies) | 162 | 10.8 | 0.1 | 0.8 | 0 | 0 | 0.1 | 1.3 | 0.4 | 3.8 | 0.9 | 8.4 | 1.0 | 9.3 | | HeLa cells (2.5 cells) | 162 | 12.4 | 0 | 0 | 0.4 | 3.3 | 0.4 | 3.1 | 0 | 0 | 2.3 | 18.4 | 2.4 | 19.0 | | SiHa cells (1 cell) | 162 | 7.5 | 0.3 | 3.7 | 1.0 | 13.0 | 0 | 0 | 0 | 0 | 4.8 | 63.6 | 4.9 | 65.0 | SD = standard deviation; CV = coefficient of variation; IVT = in vitro transcript; S/CO = signal to cutoff ratio ^One sample had an invalid APTIMA HPV Assay result and was not included in the analyses. Note: Variability from some factors may be numerically negative. This can occur if the variability due to those factors is very small. In these cases, SD and CV are shown as 0. APTIMA HPV Assay Reproducibility Study 2: Signal Variability for Panel Members with Expected Positive Results | Panel Description (copies or cells/reaction) | n | Mean S/CO | Between Instruments | | Between Operators | | Between Lots | | Between Worklists | | Within Worklists | | Total | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | | | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | | HPV 16 & HPV 18 IVT (100 copies) | 162 | 23.2 | 0.4 | 1.5 | 0.6 | 2.3 | 0.8 | 3.4 | 0.8 | 3.4 | 1.5 | 6.3 | 2.0 | 8.4 | | SiHa cells (3 cells) & HeLa cells (7.5 cells) | 162 | 18.6 | 0 | 0 | 1.7 | 9.3 | 0 | 0 | 3.5 | 18.6 | 3.7 | 20.0 | 5.4 | 28.9 | | HPV 18 IVT (100 copies) | 160 | 11.9 | 0.1 | 0.6 | 0.2 | 1.6 | 0.8 | 7.0 | 0.4 | 3.6 | 1.3 | 11.3 | 1.7 | 13.8 | | HPV 16 IVT (100 copies) | 162 | 10.8 | 0 | 0 | 0.1 | 1.3 | 0 | 0 | 0.2 | 2.2 | 0.7 | 6.1 | 0.7 | 6.6 | | MS751 cells (1 cell) | 162 | 13.6 | 0 | 0 | 0.6 | 4.3 | 0 | 0 | 2.5 | 18.4 | 2.1 | 15.2 | 3.3 | 24.2 | | ME180 cells (0.3 cells) | 162 | 5.8 | 0 | 0 | 0.6 | 10.8 | 0.5 | 9.4 | 2.2 | 36.9 | 1.7 | 29.7 | 2.9 | 49.5 | | HPV 18 IVT (30 copies) | 162 | 8.8 | 0.4 | 4.4 | 0.5 | 6.0 | 0.7 | 7.9 | 1.0 | 11.5 | 1.9 | 21.4 | 2.4 | 26.6 | PMA P100042: FDA Summary of Safety and Effectiveness Data {9} | HPV 16 IVT (30 copies) | 162 | 10.5 | 0 | 0 | 0.1 | 1.3 | 0.2 | 2.0 | 1.6 | 14.9 | 1.2 | 11.2 | 2.0 | 18.8 | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | HeLa cells (2.5 cells) | 159 | 12.0 | 0.6 | 5.1 | 1.0 | 8.5 | 0 | 0 | 2.8 | 23.8 | 2.0 | 16.6 | 3.7 | 30.6 | | SiHa cells (1 cell) | 162 | 7.4 | 0.9 | 12.5 | 0 | 0 | 0.7 | 9.3 | 1.8 | 24 | 4.2 | 56.8 | 4.7 | 63.8 | SD = standard deviation; CV = coefficient of variation; IVT = in vitro transcript; S/CO = signal to cutoff ratio ^Five samples had invalid APTIMA HPV Assay results (2 for HPV 18 IVT (100 copies), 3 for HeLa cells (2.5 cells)) and were not included in the analyses. Note: Variability from some factors may be numerically negative. This can occur if the variability due to those factors is very small. In these cases, SD and CV are shown as 0. A third study was also conducted to determine assay reproducibility by testing a 6-member panel of pooled clinical PreservCyt specimens. Six unique pools of residual HPV-negative ThinPrep liquid cytology specimens were prepared as the matrix, two of which were tested as HPV-negative panel members. Four unique pools of HPV-positive ThinPrep liquid cytology specimens were used to prepare the low (n=2) and high (n=2) HPV-positive panel members. The low positive panel members had concentrations at the limit of detection of the assay (expected positivity: ≥ 95% determined for each individual HPV-positive pool from testing serial dilutions of the pools). The high positive panel members had concentrations at 1-2 logs above the estimated limit of detection for each individual HPV positive pool (expected positivity: 100% positivity). Each panel member was transferred (1 mL) into an APTIMA Specimen Transfer tube containing STM on the day of testing. Testing was conducted in-house by 2 operators using 1 reagent lot, 3 instruments, over 6 days (3 days for each operator), testing 2 runs per day in which the panel was tested in duplicate. The panel members are described below, along with a summary of the agreement with expected results and analyte S/CO values at the 2.5th, 50th, and 97.5th percentiles of the signal distribution. APTIMA HPV Assay Reproducibility Study 3: Panel Description, Percent Agreement, and Percentile Distribution of Analyte S/CO Values | Panel Description | % agreement (95% CI) | Analyte S/CO Percentile | | | | --- | --- | --- | --- | --- | | | | 2.5^{th} | 50^{th} | 97.5^{th} | | Low positive 1 | 98.6 (71/72) (92.5, 99.8) | 1.5 | 10.1 | 19.3 | | Low positive 2 | 100 (72/72) (94.9, 100) | 1.5 | 10.3 | 19.1 | | High positive 1 | 100 (72/72) (94.9, 100) | 12.6 | 23.1 | 32.4 | | High positive 2 | 100 (72/72) (94.9, 100) | 13.3 | 24.7 | 31.2 | | Negative 1 | 98.6 (71/72) (92.5, 99.8) | 0 | 0 | 0.3 | | Negative 2 | 94.4 (68/72) (86.6, 97.8) | 0 | 0 | 0.7 | PMA P100042: FDA Summary of Safety and Effectiveness Data {10} The analyte S/CO variability for the panel members with expected positive results is shown in the table below. ## APTIMA HPV Assay Reproducibility Study 3: Signal Analysis for Panel Members with Expected Positive Results | Panel Description | n | Mean S/CO | Between Instruments | | Between Operators | | Between Lots | | Between Worklists | | Within Worklists | | Total | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | | | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | SD | CV (%) | | Low positive 1 | 72 | 9.8 | 0 | 0 | 0 | 0 | 0 | 0 | 2.2 | 22.8 | 3.0 | 30.4 | 3.7 | 38.0 | | Low positive 2 | 72 | 10.5 | 0 | 0 | 2.2 | 21.0 | 0.9 | 9.0 | 3.7 | 35.3 | 2.7 | 26.1 | 5.2 | 49.5 | | High positive 1 | 72 | 22.7 | 1.3 | 5.6 | 0 | 0 | 0.1 | 0.5 | 3.0 | 13.3 | 3.7 | 16.4 | 5.0 | 21.9 | | High positive 2 | 72 | 23.9 | 0 | 0 | 0 | 0 | 0 | 0 | 2.9 | 12.3 | 3.0 | 12.4 | 4.2 | 17.4 | SD = standard deviation; CV = coefficient of variation; S/CO = signal to cutoff ratio Note: Variability from some factors may be numerically negative. This can occur if the variability due to those factors is very small. In these cases, SD and CV are shown as 0. ## Cross-Reactivity The analytical specificity of the APTIMA HPV Assay was evaluated with PreservCyt solution media diluted 1:2.9 into STM and spiked with cultured bacteria, yeast, or fungi; cultured virus; or low-risk HPV in vitro transcripts. The organisms and test concentrations are identified in the table below. The study criteria for assessing the effect of the presence of microorganism on the specificity of the assay were based on positivity. Cross-reactivity was observed with low-risk HPV genotypes 26, 67, 70, and 82, but not with any of the other organisms tested. ### Analytical Specificity Panel: Organisms and Concentration with No Cross-Reactivity | Organism | Test Concentration with No Cross-Reactivity | Organism | Test Concentration with No Cross-Reactivity | | --- | --- | --- | --- | | Bacteria | | | | | Acinetobacter lwoffii | 1x10^{8} CFU/mL | Listeria monocytogenes | 1x10^{8} CFU/mL | | Actinomyces israelii | 1x10^{8} CFU/mL | Micrococcus luteus | 1x10^{8} CFU/mL | | Alcaligenes faecalis | 1x10^{8} CFU/mL | Mobiluncus curtisii | 2x10^{7} CFU/mL | | Atopobium vaginae | 5x10^{7} CFU/mL | Mycobacterium smegmatis | 1x10^{8} CFU/mL | | Bacillus cereus | 1x10^{8} CFU/mL | Mycoplasma fermentans | 5x10^{7} CFU/mL | | Bacteroides fragilis | 1x10^{8} CFU/mL | Mycoplasma genitalium | 1x10^{8} CFU/mL | | Bacteroides ureolyticus | 1x10^{8} CFU/mL | Mycoplasma hominis | 5x10^{7} CFU/mL | | Bifidobacterium adolescentis | 1x10^{8} CFU/mL | Neisseria gonorrhoeae | 1x10^{8} CFU/mL | | Bifidobacterium breve | 1x10^{8} CFU/mL | Neisseria gonorrhoeae and Chlamydia trachomatis | 2.5x10^{7} CFU/mL 2.3x10^{3} TCID_{50}/mL | | Campylobacter fetus-fetus | 1x10^{8} CFU/mL | Neisseria meningitidis | 1x10^{8} CFU/mL | | Chlamydia trachomatis | 3.2x10^{3} TCID_{50}/mL | Peptoniphilus lacrimalis | 1x10^{8} CFU/mL | PMA P100042: FDA Summary of Safety and Effectiveness Data {11} PMA P100042: FDA Summary of Safety and Effectiveness Data page 12 | Organism | Test Concentration with No Cross-Reactivity | Organism | Test Concentration with No Cross-Reactivity | | --- | --- | --- | --- | | Clostridium difficile | 6x10^{7} CFU/mL | Peptostreptococcus anaerobius | 1x10^{8} CFU/mL | | Clostridium perfringens | 1x10^{8} CFU/mL | Propionibacterium acnes | 1x10^{8} CFU/mL | | Corynebacterium genitalium | 1x10^{8} CFU/mL | Proteus mirabilis | 1x10^{8} CFU/mL | | Corynebacterium xerosis | 1x10^{8} CFU/mL | Proteus vulgaris | 1x10^{8} CFU/mL | | Enterobacter cloacae | 1x10^{8} CFU/mL | Providencia stuartii | 1x10^{8} CFU/mL | | Enterococcus faecalis | 1x10^{8} CFU/mL | Pseudomonas aeruginosa | 1x10^{8} CFU/mL | | Escherichia coli | 1x10^{8} CFU/mL | Ruminococcus productus | 1x10^{8} CFU/mL | | Finegoldia magna | 1x10^{8} CFU/mL | Serratia marcescens | 1x10^{8} CFU/mL | | Fusobacterium nucleatum | 1x10^{8} CFU/mL | Staphylococcus aureus | 1x10^{8} CFU/mL | | Gardnerella vaginalis | 1x10^{8} CFU/mL | Staphylococcus epidermidis | 1x10^{8} CFU/mL | | Haemophilus ducreyi | 1x10^{8} CFU/mL | Staphylococcus saprophyticus | 1x10^{8} CFU/mL | | Klebsiella pneumoniae | 1x10^{8} CFU/mL | Streptococcus agalactiae | 1x10^{8} CFU/mL | | Lactobacillus acidophilus | 1x10^{8} CFU/mL | Streptococcus pyogenes | 1x10^{8} CFU/mL | | Lactobacillus crispatus | 1x10^{8} CFU/mL | Streptococcus sanguinis | 1x10^{8} CFU/mL | | Lactobacillus delbrueckii ssp. bulgaricus | 1x10^{8} CFU/mL | Ureaplasma urealyticum | 1x10^{8} CFU/mL | | Lactobacillus jensenii | 1x10^{8} CFU/mL | | | | Yeast/protozoa | | | | | Candida albicans | 1x10^{8} CFU/mL | Trichomonas vaginalis | 1x10^{7} cells/mL | | Viruses | | | | | Adenovirus 2 | 1x10^{7} vp/mL | Herpes simplex virus 1 | 2.5x10^{5} TCID_{50}/mL | | Cytomegalovirus | 5.6x10^{5} TCID_{50}/mL | Herpes simplex virus 2 | 5x10^{4} TCID_{50}/mL | | Epstein-Barr virus | 4.3x10^{6} vp/mL | SV40 | 1.2 x10^{4} TCID_{50}/mL | | HIV-1 | 1.0x10^{6} copies/mL | | | | Non-targeted HPV genotypes | | | | | HPV 6 | 2.5x10^{6} copies/mL | HPV 61 | 2.5x10^{6} copies/mL | | HPV 11 | 2.5x10^{6} copies/mL | HPV 67 | 1 copy/mL | | HPV 26 | 2.5 copies/mL | HPV 69 | 2.5x10^{6} copies/mL | | HPV 30 | 2.5x10^{6} copies/mL | HPV 70 | 1 copy/mL | | HPV 34 | 2.5x10^{6} copies/mL | HPV 71 | 2.5x10^{6} copies/mL | | HPV 42 | 2.5x10^{6} copies/mL | HPV 73 | 2.5x10^{6} copies/mL | | HPV 43 | 2.5x10^{6} copies/mL | HPV 81 | 2.5x10^{6} copies/mL | | HPV 44 | 2.5x10^{6} copies/mL | HPV 82 | 2.5 copies/mL | | HPV 53 | 2.5x10^{6} copies/mL | HPV 85 | 2.5x10^{6} copies/mL | {12} | HPV 54 | 2.5x10^{6} copies/mL | | | | --- | --- | --- | --- | vp = viral particles CFU = colony forming units TCID50 = tissue culture infective dose 50 Note: Bold indicates types where cross-reactivity (> 5% positivity) was observed when tested at concentrations greater than that noted in the table. The analytical sensitivity of the APTIMA HPV Assay in the presence of microorganisms was evaluated with the same panel described directly above, which was also spiked with a low concentration of HPV infected SiHa cells (1 cell per reaction). The study criteria for assessing the effect of the presence of microorganism on the sensitivity of the assay were based on positivity. The sensitivity of the APTIMA HPV Assay was not affected by any of the organisms tested. ## Interference The substances described in the table below were individually spiked into PreservCyt solution at 1% and 10% v/v or w/v, diluted with STM and then tested in the APTIMA HPV Assay. All substances were tested in the presence and absence of HPV infected cultured cells (SiHa, 3 cells/reaction). Interference was observed with two of the seven lubricants that contained Polyquaternium 15, and one of the five anti-fungal medications that contained tioconazole. Interference was not observed with any of other substances tested. ### Substances Tested for Possible Interference with the APTIMA HPV Assay | Product Category | Product Brand or Type | Highest Concentration* Tested that Did Not Interfere with Assay Performance | | --- | --- | --- | | Lubricant | KY Sensual Mist | 10% v/v | | | KY Warming Jelly | 10% w/v | | | KY Warming Liquid | 10% v/v | | | CVS Brand Personal Lubricant | 10% w/v | | | Target Brand Warming Massage Lotion and Personal Lubricant | 10% v/v | | | Astroglide Personal Lubricant | 0.3% w/v (0.075% w/v test sample) | | | Target Brand Lubricating Liquid | 0.1% v/v (0.025% v/v test sample) | | Spermicide | Gynol II Vaginal Contraceptive Original Formula | 10% w/v | | | Gynol II Vaginal Contraceptive Extra Strength | 10% w/v | | | Delfen Vaginal Contraceptive Foam | 10% w/v | | | Encare Vaginal Contraceptive | 10% w/v | | | Conceptrol Vaginal Contraceptive | 10% w/v | | Anti-fungal/ Anti-Iteh Medication | Vagisil Maximum Strength | 10% w/v | | | Monistat Soothing Care | 10% w/v | | | Monistat 3 Combination Pack | 10% w/v | PMA P100042: FDA Summary of Safety and Effectiveness Data {13} | | Target Brand Ticonazole 1 | 0.3% w/v (0.075% w/v test sample) | | --- | --- | --- | | | Target Brand Miconazole 3 | 10% w/v | | Glacial Acetic Acid | EMD M/N AX0073-11 | 10% v/v | | Whole Blood | whole blood | 10% v/v | ## Reagent Stability Expiration dating for this device has been established and approved at 18 months for the APTIMA HPV Assay when stored at 2 - 8°C, with the exception of the subset of reagents in the APTIMA HPV Room Temperature Box, which should be stored at 15 - 30°C. ## Sample Handling and Collection Cervical specimens should be collected in PreservCyt Solution, the ThinPrep Pap Test preservation system, using a broom-type device (e.g. Rovers Cervex Brush, Wallach Papette), or Endocervical Brush/Spatula. Specimen stability studies demonstrated that for the APTIMA HPV Assay cervical specimens should be transported and stored at 2°C to 30°C, with no more than 30 days at temperatures above 8°C. PreservCyt specimens should be transferred to an APTIMA Specimen Transfer tube within 105 days of collection. PreservCyt Solution specimens transferred to an APTIMA Specimen Transfer tube may be stored at 2°C to 30°C for up to 60 days. If longer storage is needed, the PreservCyt Solution specimen or the PreservCyt Solution specimen diluted into the Specimen Transfer tube may be stored at -20°C for up to 24 months. ## TIGRIS DTS System Carryover A study was conducted to determine the rate of false positive results observed with the Aptima HPV Assay using the TIGRIS DTS System when samples containing high titer HPV were interspersed throughout specimen processing racks containing HPV-negative samples. High titer positive samples for this study were created by spiking HPV 16 in vitro transcript into a solution comprised of 1 part PreservCyt solution and 2.9 parts specimen transport media (STM) to a concentration of 2.5×10⁶ copies/mL (1×10⁶ copies/reaction). The sample to sample cross-contamination rate of the APTIMA HPV Assay on the TIGRIS DTS System was determined to be 0.3%. ## ThinPrep Carryover Study A study was conducted to determine the false positive rate observed with the APTIMA HPV Assay when PreservCyt liquid Pap specimens containing a high concentration of spiked HPV-positive cells, were alternately processed with HPV-negative specimens on the ThinPrep 2000 Processor. The study also evaluated the effectiveness of the cleaning procedure described in the APTIMA Specimen Transfer Kit package insert. The observed false positive rate for specimens tested with the Aptima HPV Assay following processing on the TP2000 was 0.8% when the cleaning procedure described in the APTIMA Specimen Transfer Kit package insert was followed. When these instructions were not PMA P100042: FDA Summary of Safety and Effectiveness Data {14} followed, the false positive rate was higher (1.7%). Users should follow the cleaning instructions provided via the APTIMA Specimen Transfer kit for decontamination between specimens to minimize carryover risk. ## B. Animal Studies Not applicable ## C. Additional Studies Not applicable ## X. SUMMARY OF PRIMARY CLINICAL STUDIES ### A. Study Design Patients were enrolled between March 2008 and December 2009. This was a prospective cohort, multi-center trial consisting of 2 sub-studies to support the two distinct indications for use of the assay, the ASC-US Study and the NILM (negative for intraepithelial lesions or malignancy) Study. The database for this PMA included 12,896 patients age 21 and older. There were 19 enrolling (collection) sites and 3 APTIMA HPV Assay testing sites. One of the collection sites was excluded from the final analysis due to protocol violations (see Section XI below). ### 1. Clinical Inclusion and Exclusion Criteria Enrollment in the study was limited to patients who attended a participating clinic and underwent a routine cytology test. In addition, the subject must have been able to comprehend and sign an IRB-approved Informed Consent Form and other applicable study enrollment documents. To be included in the ASC-US Study, the subject's referral cytology specimen must have had ASC-US results. To be included in the NILM Study, the subject must have been ≥30 years of age and the subject's referral cytology specimen must have had NILM cytology test results. Note that all women under age 21 that were enrolled in the ASC-US study have been removed from the dataset (n=99), including patient demographics and subject accountability. The approved indication does not cover this age range since HPV testing is not recommended in women under the age of 21². Patients were not permitted to enroll in the either study if the subject, clinician, or medical record reported any of the following: - History of cervical disease (cancer or precancerous condition) in the previous 12 months - History of an abnormal cytology test result in the previous 12 months - Under 18 years of age, without the documented consent of her parent or legal guardian - Subject is known to be pregnant at enrollment - History of illness that the investigator considers could interfere with or affect the conduct, results, and/or completion of the clinical trial PMA P100042: FDA Summary of Safety and Effectiveness Data {15} - History of illness that the investigator/physician considers to create an unacceptable risk to the subject if enrolled - History of HPV vaccination prior to enrollment. ## 2. Follow-up Schedule Patients were scheduled to return for follow-up examinations as described below. ## APTIMA HPV Assay Clinical Trial Study Design A prospective, multicenter US clinical study known as the CLEAR trial was conducted to determine the clinical performance of the APTIMA HPV Assay for detection of cervical intraepithelial neoplasia grade 2 or more severe cervical disease (≥CIN2). Women were enrolled into either the ASC-US Study or the NILM Study based on cytology results from routine cervical cancer screening. The ASC-US Study population included women 21 years and older with ASC-US cytology results and the NILM Study population included women 30 years of age and older with NILM cytology results. The NILM Study was designed to support the adjunctive screening claim for women 30 years and older, since women in this age range with cytology results greater than ASC-US should proceed to colposcopy regardless of their HPV status.² Women from 18 clinical sites, primarily obstetrics/gynecology clinics, which covered a wide geographic distribution and a diverse population, were analyzed. Eligible women were assigned to the ASC-US Study or NILM Study based on their referral ThinPrep PreservCyt liquid based cytology specimen. Residual referral specimens were tested with both the APTIMA HPV Assay and an FDA-approved HPV DNA test. All women in the ASC-US Study were referred to colposcopy, regardless of their HPV test results. An endocervical curettage (ECC) biopsy and cervical punch biopsies (1 biopsy from each of the 4 quadrants) were obtained. If a lesion was visible, a punch biopsy was obtained (directed method; 1 biopsy per lesion) and quadrants without a visible lesion were biopsied at the squamocolumnar junction (random method). In the NILM Study, women positive with the APTIMA HPV Assay and/or the FDA-approved HPV DNA test, as well as randomly selected women who were negative with both assays, were referred to colposcopy for the baseline evaluation. The randomly selected women who were negative for both assays were included to correct for verification bias with adjusted performance estimates generated using a multiple imputation method. An ECC biopsy was obtained from each woman who attended colposcopy. Punch biopsies were obtained from visible lesions only (directed method; 1 biopsy per lesion). Follow-up of women in the NILM Study who do not have ≥CIN2 is ongoing for 3 years with annual cytology visits. Women with ASC-US or more severe cytology results during the follow-up period are referred to colposcopy using the same biopsy procedure performed for the baseline evaluation. For both studies, disease status was determined from a consensus histology review panel, which was based on agreement of at least 2 expert pathologists. The expert PMA P100042: FDA Summary of Safety and Effectiveness Data 21 {16} pathologists were masked to the woman’s HPV and cytology status, as well as each other’s histology diagnoses. Investigators, clinicians, and women were masked to the HPV test results until after completion of the colposcopy visit, to avoid bias. Clinical performance of the APTIMA HPV Assay was determined for detection of ≥CIN2 and cervical intraepithelial neoplasia grade 3 or more severe cervical disease (≥CIN3). Clinical performance of the FDA-approved HPV DNA test was also determined for direct comparison to the APTIMA HPV Assay results. ## B. Accountability of PMA Cohort ### Accountability in ASC-US (≥ 21 years) Population Subject enrollment began on March 19, 2008, and was completed on December 23, 2009. Colposcopy visits were completed on December 23, 2009. APTIMA HPV Assay testing was completed on January 29, 2010. There were 958 evaluable women, 21 years or older, who met the selection criteria and were enrolled into the ASC-US Study. These evaluable women had a valid APTIMA HPV Assay result and attended the colposcopy visit. Nineteen (19) of the 958 women had indeterminate disease status. The remaining 939 subjects had conclusive disease status based on the evaluation of all biopsies performed (eg, directed punch, random, and ECC) and were included in the clinical performance analyses. The ASC-US study enrollment and sample accountability are described below. ![img-0.jpeg](img-0.jpeg) PMA P100042: FDA Summary of Safety and Effectiveness Data page 17 {17} # Accountability in NILM (≥ 30 years) population Subject enrollment began on March 31, 2008, and was completed on December 8, 2009. Baseline colposcopy visits were completed on February 2, 2010. APTIMA HPV Assay testing was completed on January 28, 2010. Follow-up is currently ongoing and is estimated to be completed by April 2013. There were 10,871 evaluable women 30 years of age and older with NILM cytology results and APTIMA HPV Assay results. Of the 540 women with positive APTIMA HPV Assay results, 335 attended colposcopy. Of the 10,331 women with negative APTIMA HPV Assay results, 530 attended colposcopy (865 total). The NILM Study enrollment and sample accountability are described below. ![img-1.jpeg](img-1.jpeg) PMA P100042: FDA Summary of Safety and Effectiveness Data page 18 {18} # C. Study Population Demographics and Baseline Parameters The demographics of the study population are typical for a prospective study performed in the US. See tables below for detailed demographic information on each study population. | ASC-US Subject Demographics (N=958) | | | --- | --- | | Age (years) at consent | | | Mean | 33.8 | | SD | 10.3 | | Median | 31.0 | | Min | 21 | | Max | 85 | | | | | Race | | | White | 587 | | Black or African American | 229 | | Asian | 29 | | American Indian or Alaska Native | 34 | | Native Hawaiian or Pacific Islander | 1 | | Unknown / Refused | 106 | | | | | Ethnicity | | | Hispanic or Latino | 195 | | Not Hispanic or Latino | 712 | | Unknown / Refused | 51 | | NILM Subject Demographics (N=10871) | | | --- | --- | | Age (years) at consent | | | Mean | 44.1 | | SD | 10.1 | | Median | 43.0 | | Min | 30 | | Max | 89 | | | | | Race | | | White | 6937 | | Black or African American | 1390 | | Asian | 658 | | American Indian or Alaska Native | 152 | | Native Hawaiian or Pacific Islander | 66 | | Unknown / Refused | 1808 | | | | | Ethnicity | | | Hispanic or Latino | 3421 | | Not Hispanic or Latino | 6881 | | Unknown / Refused | 569 | PMA P100042: FDA Summary of Safety and Effectiveness Data {19} # D. Safety and Effectiveness Results ASC-US ≥ 21 Years Population: APTIMA HPV Assay Clinical Performance In total, there were 1252 women 21 years of age and older with ASC-US cytology results enrolled in the ASC-US Study. Of these, 294 women were withdrawn and 19 had an undetermined disease diagnosis; all were excluded from analysis. The remaining 939 evaluable women were 21 years of age and older with ASC-US cytology results, APTIMA HPV Assay results, and conclusive disease status. Ninety-one (91) women had ≥CIN2 and forty-one (41) had ≥CIN3. Prevalence of ≥CIN2 and ≥CIN3 in evaluable women with ASC-US cytology results were 9.7% and 4.4%, respectively. The results of the APTIMA HPV Assay by the consensus histology review panel diagnoses are presented below. ASC-US ≥ 21 Years Population: Results of the APTIMA HPV Assay by Consensus Histology Review Panel Diagnosis | APTIMA HPV Assay Result* | HPV DNA Test | Consensus Histology Review Panel Diagnosis | | | | | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | | Undetermined** | Normal | CIN1 | CIN2 | CIN3 | Cancer | Total | | Positive | Positive | 6 | 170 | 113 | 41 | 32 | 1 | 363 | | Positive | Negative | 0 | 7 | 0 | 1 | 2 | 0 | 10 | | Positive | No Result*** | 0 | 14 | 11 | 0 | 2 | 0 | 27 | | Negative | Positive | 0 | 47 | 13 | 2 | 3 | 0 | 65 | | Negative | Negative | 10 | 371 | 55 | 6 | 1 | 0 | 443 | | Negative | No Result*** | 3 | 40 | 7 | 0 | 0 | 0 | 50 | | Total | | 19 | 649 | 199 | 50 | 40 | 1*** | 958 | *All samples had final valid results (upon initial testing or after resolution of initial invalids per procedure). **19 subjects attended the colposcopy visit but a diagnosis could not be determined for the following reasons: < 5 biopsy specimens obtained all with histology results of Normal/CIN1 (n=15), no biopsies collected (n=3), and biopsy slides lost (n=1). ***77 women with APTIMA HPV Assay results did not have HPV DNA test results primarily due to insufficient volume of the cytology specimen. ***One subject had adenocarcinoma in situ (AIS). Clinical performance estimates of the APTIMA HPV Assay including sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) for the detection of ≥CIN2 and ≥CIN3 based on evaluating all biopsies and including only directed biopsies are shown in the table below, as are the estimates for the FDA-approved HPV DNA test. Ninety-five percent two-sided confidence intervals for sensitivity and specificity were calculated using a score method³ and 95% two-sided confidence intervals for the PPV and NPV were calculated based on the confidence intervals of corresponding likelihood ratios (positive likelihood ratio for PPV and negative likelihood ratio for NPV).⁴,⁵ PMA P100042: FDA Summary of Safety and Effectiveness Data 25 {20} ASC-US ≥ 21 Years Population: Performance of the APTIMA HPV Assay and an FDA-approved HPV DNA Test for Detection of ≥CIN2 and ≥CIN3 | | Performance | APTIMA HPV Assay N=939 | | HPV DNA Test N=865* | | | --- | --- | --- | --- | --- | --- | | | | Estimate | (95% CI) | Estimate | (95% CI) | | ≥CIN2 | All Biopsies | | | | | | | Sensitivity (%) | 86.8 (79/91) | (78.4, 92.3) | 88.8 (79/89) | (80.5, 93.8) | | | Specificity (%) | 62.9 (533/848) | (59.6, 66.0) | 55.8 (433/776) | (52.3, 59.3) | | | PPV (%) | 20.1 (79/394) | (18.1, 22.0) | 18.7 (79/422) | (17.0, 20.4) | | | NPV (%) | 97.8 (533/545) | (96.5, 98.8) | 97.7 (433/443) | (96.2, 98.8) | | | Prevalence (%) | 9.7 (91/939) | | 10.3 (89/865) | | | | Directed Biopsies** | | | | | | | Sensitivity (%) | 93.3 (56/60) | (84.1, 97.4) | 93.2 (55/59) | (83.8, 97.3) | | | Specificity (%) | 61.5 (539/876) | (58.3, 64.7) | 54.5 (438/804) | (51.0, 57.9) | | | PPV (%) | 14.2 (56/393) | (12.7, 15.6) | 13.1 (55/421) | (11.7, 14.2) | | | NPV (%) | 99.3 (539/543) | (98.3, 99.8) | 99.1 (438/442) | (97.9, 99.7) | | | Prevalence (%) | 6.4 (60/936) | | 6.8 (59/863) | | | ≥CIN3 | All Biopsies | | | | | | | Sensitivity (%) | 90.2 (37/41) | (77.5, 96.1) | 92.3 (36/39) | (79.7, 97.3) | | | Specificity (%) | 60.2 (541/898) | (57.0, 63.4) | 53.3 (440/826) | (49.9, 56.6) | | | PPV (%) | 9.4 (37/394) | (8.1, 10.4) | 8.5 (36/422) | (7.4, 9.4) | | | NPV (%) | 99.3 (541/545) | (98.3, 99.8) | 99.3 (440/443) | (98.3, 99.8) | | | Prevalence (%) | 4.4 (41/939) | | 4.5 (39/865) | | | | Directed Biopsies** | | | | | | | Sensitivity (%) | 93.1 (27/29) | (78.0, 98.1) | 96.4 (27/28) | (82.3, 99.4) | | | Specificity (%) | 59.6 (541/908) | (56.4, 62.7) | 52.8 (441/836) | (49.4, 56.1) | | | PPV (%) | 6.9 (27/394) | (5.8, 7.6) | 6.4 (27/422) | (5.5, 7.0) | | | NPV (%) | 99.6 (541/543) | (98.8, 100) | 99.8 (441/442) | (98.9, 100) | | | Prevalence (%) | 3.1 (29/937) | | 3.2 (28/864) | | *74 women with APTIMA HPV Assay results did not have HPV DNA Test results primarily due to insufficient volume of the cytology specimen. PMA P100042: FDA Summary of Safety and Effectiveness Data {21} **Consensus histology result was derived using only results from directed biopsies. Women with no directed biopsies reflect a normal colposcopy and are included in these analyses as non-diseased (<CIN2 or <CIN3, as appropriate). A consensus was not always reached when only directed biopsies were included. There were 2 women excluded from both the ≥CIN2 and ≥CIN3 analysis based on evaluation of only directed biopsy slides: 1 woman had normal histology by one pathologist and undetermined by the second pathologist and the other woman had undetermined histology by both pathologists. One additional woman was excluded from the ≥CIN2 analysis based on evaluation of only directed biopsy slides because she was CIN1 by one pathologist and CIN2 by the second pathologist. She was included in the ≥CIN3 analysis as disease negative.** When evaluating all biopsies, clinical sensitivity estimates of the APTIMA HPV Assay and the FDA-approved HPV DNA test, where both assay results are available for the detection of ≥CIN2 and ≥CIN3, were similar (differences in sensitivity estimates were not statistically significant: sensitivity difference = -2.3% [95% CI: -9.5%, 4.8%]). Clinical specificity estimates of the APTIMA HPV Assay for the detection of ≥CIN2 and ≥CIN3 were higher than those of the FDA-approved HPV DNA test (differences in specificity estimates were statistically significant). For ≥CIN2, the specificity difference was 6.8% (95% CI: 4.9%, 9.0%). NPVs were similar but for the detection of ≥CIN2, the PPV for the APTIMA HPV Assay was slightly higher than PPV for the FDA-approved HPV DNA test (20.1% vs 18.7%). Of the 91 ≥CIN2 cases, 60 (65.9%) were identified in directed biopsies and 31 (34.1%) were identified from random and/or ECC biopsies (i.e., not in directed biopsies). These findings are comparable to results from published studies, in which approximately 25% to 40% of ≥CIN2 cases were identified from random and/or ECC biopsy specimens only.⁶,⁷ Using only directed biopsies to determine disease status (assuming women with no directed biopsies had normal histology results because no visible lesions were present), prevalence of ≥CIN2 and ≥CIN3 in the study were 6.4% and 3.1%, respectively. The clinical sensitivity estimates for the detection of ≥CIN2 and ≥CIN3 were higher for both tests using directed biopsies only than estimates calculated using all biopsies. For both assays, clinical specificity using only directed biopsies was similar to the specificity obtained with all biopsies included. Accordingly, when using only directed biopsies, the APTIMA HPV Assay specificity was significantly higher than that of the FDA-approved HPV DNA test. PMA P100042: FDA Summary of Safety and Effectiveness Data page 22 27 {22} Clinical performance estimates of the APTIMA HPV Assay and the FDA-approved HPV DNA test are shown by age group in the tables below (≥CIN2 and ≥CIN3, respectively, based on evaluating all biopsies). ## ASC-US ≥ 21 Years Population: Performance of the APTIMA HPV Assay and an FDA-approved HPV DNA Test for Detection of ≥CIN2 by Age Group | | Performance | APTIMA HPV Assay N=939 | | HPV DNA test N=865* | | | --- | --- | --- | --- | --- | --- | | | | Estimate | (95% CI) | Estimate | (95% CI) | | 21 to 29 Years | | N=415 | | N=389 | | | | Sensitivity (%) | 90.2 (55/61) | (80.2, 95.4) | 94.9 (56/59) | (86.1, 98.3) | | | Specificity (%) | 44.9 (159/354) | (39.8, 50.1) | 35.5 (117/330) | (30.5, 40.8) | | | PPV (%) | 22.0 (55/250) | (19.6, 24.2) | 20.8 (56/269) | (19.0, 22.5) | | | NPV (%) | 96.4 (159/165) | (93.0, 98.5) | 97.5 (117/120) | (93.6, 99.4) | | | Prevalence (%) | 14.7 (61/415) | | 15.2 (59/389) | | | 30 to 39 Years | | N=262 | | N=239 | | | | Sensitivity (%) | 90.0 (18/20) | (69.9, 97.2) | 80.0 (16/20) | (58.4, 91.9) | | | Specificity (%) | 68.2 (165/242) | (62.1, 73.7) | 61.6 (135/219) | (55.1, 67.8) | | | PPV (%) | 18.9 (18/95) | (14.7, 22.7) | 16.0 (16/100) | (11.8, 19.6) | | | NPV (%) | 98.8 (165/167) | (96.5, 99.8) | 97.1 (135/139) | (94.1, 99.1) | | | Prevalence (%) | 7.6 (20/262) | | 8.4 (20/239) | | | ≥ 40 Years | | N=262 | | N=237 | | | | Sensitivity (%) | 60.0 (6/10) | (31.3, 83.2) | 70.0 (7/10) | (39.7, 89.2) | | | Specificity (%) | 82.9 (209/252) | (77.8, 87.1) | 79.7 (181/227) | (74.0, 84.4) | | | PPV (%) | 12.2 (6/49) | (5.8, 18.4) | 13.2 (7/53) | (6.9, 18.7) | | | NPV (%) | 98.1 (209/213) | (96.6, 99.4) | 98.4 (181/184) | (96.6, 99.6) | | | Prevalence (%) | 3.8 (10/262) | | 4.2 (10/237) | | *74 women with APTIMA HPV Assay results did not have HPV DNA Test results primarily due to insufficient volume of the cytology specimen. PMA P100042: FDA Summary of Safety and Effectiveness Data {23} ASC-US $\geq 21$ Years Population: Performance of the APTIMA HPV Assay and an FDA-approved HPV DNA Test for Detection of $\geq$ CIN3 by Age Group | | Performance | APTIMA HPV Assay N=939 | | HPV DNA test N=865* | | | --- | --- | --- | --- | --- | --- | | | | Estimate | (95% CI) | Estimate | (95% CI) | | 21 to 29 Years | | N=415 | | N=389 | | | | Sensitivity (%) | 96.3 (26/27) | (81.7, 99.3) | 100 (25/25) | (86.7, 100) | | | Specificity (%) | 42.3 (164/388) | (37.5, 47.2) | 33.0 (120/364) | (28.3, 38.0) | | | PPV (%) | 10.4 (26/250) | (8.9, 11.4) | 9.3 (25/269) | (8.2, 10.0) | | | NPV (%) | 99.4 (164/165) | (97.2, 100) | 100 (120/120) | (97.5, 100) | | | Prevalence (%) | 6.5 (27/415) | | 6.4 (25/389) | | | 30 to 39 Years | | N=262 | | N=239 | | | | Sensitivity (%) | 88.9 (8/9) | (56.5, 98.0) | 77.8 (7/9) | (45.3, 93.7) | | | Specificity (%) | 65.6 (166/253) | (59.6, 71.2) | 59.6 (137/230) | (53.1, 65.7) | | | PPV (%) | 8.4 (8/95) | (5.2, 10.4) | 7.0 (7/100) | (3.9, 9.1) | | | NPV (%) | 99.4 (166/167) | (97.6, 100) | 98.6 (137/139) | (96.4, 99.8) | | | Prevalence (%) | 3.4 (9/262) | | 3.8 (9/239) | | | ≥ 40 Years | | N=262 | | N=237 | | | | Sensitivity (%) | 60.0 (3/5) | (23.1, 88.2) | 80.0 (4/5) | (37.6, 96.4) | | | Specificity (%) | 82.1 (211/257) | (77.0, 86.3) | 78.9 (183/232) | (73.2, 83.6) | | | PPV (%) | 6.1 (3/49) | (1.6, 10.2) | 7.5 (4/53) | (2.9, 10.7) | | | NPV (%) | 99.1 (211/213) | (98.0, 99.9) | 99.5 (183/184) | (98.2, 100) | | | Prevalence (%) | 1.9 (5/262) | | 2.1 (5/237) | | *74 women with APTIMA HPV Assay results did not have HPV DNA Test results primarily due to insufficient volume of the cytology specimen. PMA P100042: FDA Summary of Safety and Effectiveness Data {24} The absolute risk of disease $(\geq \mathrm{CIN2}$ and $\geq \mathrm{CIN3}$, based on evaluating all biopsies) by APTIMA HPV Assay result and the relative risk of disease for positive versus negative APTIMA HPV Assay results are shown in the table below, as are the estimates for the FDA-approved HPV DNA test. The relative risk of $\geq \mathrm{CIN2}$ was 9.1 (95% CI: 5.0, 16.5), indicating that a woman who was APTIMA HPV Assay positive was 9.1 times as likely to have $\geq \mathrm{CIN2}$ than a woman who was APTIMA HPV Assay negative. The relative risk of $\geq \mathrm{CIN3}$ was 12.8 (95% CI: 4.6, 35.6). ASC-US $\geq 21$ Years Population: Absolute and Relative Risks of $\geq$ CIN2 and $\geq$ CIN3 for Results of the APTIMA HPV Assay and an FDA-approved HPV DNA Test | | Assay Result | APTIMA HPV Assay N=939 | | HPV DNA test N=865* | | | --- | --- | --- | --- | --- | --- | | | | Absolute Risk (95% CI) | Relative Risk (95% CI) | Absolute Risk (95% CI) | Relative Risk (95% CI) | | ≥CIN2 | Positive | 20.1 (79/394) (18.1, 22.0) | 9.1 (5.0, 16.5) | 18.7 (79/422) (17.0, 20.4) | 8.3 (4.4, 15.8) | | | Negative | 2.2 (12/545) (1.2, 3.5) | | 2.3 (10/443) (1.2, 3.8) | | | | Prevalence (%) | 9.7 (91/939) | | 10.3 (89/865) | | | ≥CIN3 | Positive | 9.4 (37/394) (8.1, 10.4) | 12.8 (4.6, 35.6) | 8.5 (36/422) (7.4, 9.4) | 12.6 (3.9, 40.6) | | | Negative | 0.7 (4/545) (0.2, 1.7) | | 0.7 (3/443) (0.2, 1.7) | | | | Prevalence (%) | 4.4 (41/939) | | 4.5 (39/865) | | *74 women with APTIMA HPV Assay results did not have HPV DNA Test results primarily due to insufficient volume of the cytology specimen. PMA P100042: FDA Summary of Safety and Effectiveness Data {25} Absolute and relative risk estimates of disease (≥CIN2 and ≥CIN3, based on evaluating all biopsies) for the APTIMA HPV Assay and the FDA-approved HPV DNA test are shown by age group in the table below. ASC-US ≥ 21 Years Population: Absolute and Relative Risks of ≥CIN2 and ≥CIN3 for Results of the APTIMA HPV Assay and an FDA-approved HPV DNA Test by Age Group | | Age | Assay Result | APTIMA HPV Assay N=939 | | HPV DNA test N=865* | | | --- | --- | --- | --- | --- | --- | --- | | | | | Absolute Risk (95% CI) | Relative Risk (95% CI) | Absolute Risk (95% CI) | Relative Risk (95% CI) | | ≥CIN2 | 21 to 29 Years | | N=415 | | N=389 | | | | | Positive | 22.0 (55/250) (19.6, 24.2) | 6.1 (2.7, 13.7) | 20.8 (56/269) (19.0, 22.5) | 8.3 (2.7, 26.1) | | | | Negative | 3.6 (6/165) (1.5, 7.0) | | 2.5 (3/120) (0.6, 6.4) | | | | | Prevalence (%) | 14.7 (61/415) | | 15.2 (59/389) | | | | 30 to 39 Years | | N=262 | | N=239 | | | | | Positive | 18.9 (18/95) (14.7, 22.7) | 15.8 (3.8, 66.7) | 16.0 (16/100) (11.8, 19.6) | 5.6 (1.9, 16.1) | | | | Negative | 1.2 (2/167) (0.2, 3.5) | | 2.9 (4/139) (0.9, 5.9) | | | | | Prevalence (%) | 7.6 (20/262) | | 8.4 (20/239) | | | | ≥40 Years | | N=262 | | N=237 | | | | | Positive | 12.2 (6/49) (5.8, 18.4) | 6.5 (1.9, 22.2) | 13.2 (7/53) (6.9, 18.7) | 8.1 (2.2, 30.2) | | | | Negative | 1.9 (4/213) (0.6, 3.4) | | 1.6 (3/184) (0.4, 3.4) | | | | | Prevalence (%) | 3.8 (10/262) | | 4.2 (10/237) | | | ≥CIN3 | 21 to 29 Years | | N=415 | | N=389 | | | | | Positive | 10.4 (26/250) (8.9, 11.4) | 17.2 (2.4, 125) | 9.3 (25/269) (8.2, 10.0) | Not calculable | | | | Negative | 0.6 (1/165) (0.0, 2.8) | | 0.0 (0/120) (0.0, 2.5) | | | | | Prevalence (%) | 6.5 (27/415) | | 6.4 (25/389) | | | | 30 to 39 Years | | N=262 | | N=239 | | | | | Positive | 8.4 (8/95) (5.2, 10.4) | 14.1 (1.8, 111) | 7.0 (7/100) (3.9, 9.1) | 4.9 (1.0, 22.9) | | | | Negative | 0.6 (1/167) (0.0, 2.4) | | 1.4 (2/139) (0.2, 3.6) | | | | | Prevalence (%) | 3.4 (9/262) | | 3.8 (9/239) | | | | ≥40 Years | | N=262 | | N=237 | | | | | Positive | 6.1 (3/49) (1.6, 10.2) | 6.5 (1.1, 38.0) | 7.5 (4/53) (2.9, 10.7) | 13.9 (1.6, 122) | | | | Negative | 0.9 (2/213) (0.1, 2.0) | | 0.5 (1/184) (0.0, 1.8) | | | | | Prevalence (%) | 1.9 (5/262) | | 2.1 (5/237) | | *74 women with APTIMA HPV Assay results did not have HPV DNA Test results primarily due to insufficient volume of the cytology specimen. PMA P100042: FDA Summary of Safety and Effectiveness Data {26} # NILM ≥ 30 Years Population (NILM Study): APTIMA HPV Assay Clinical Performance In total, there were 11,644 women with NILM cytology results enrolled in the NILM Study. Of these, 773 women were withdrawn and excluded from analysis. The remaining 10,871 evaluable women were 30 years of age and older with NILM cytology results and APTIMA HPV Assay results. Of the 540 women with positive APTIMA HPV Assay results, 335 attended colposcopy. Of the 10,331 women with negative APTIMA HPV Assay results, 530 attended colposcopy. Twenty (20) women had ≥CIN2 and eleven (11) had ≥CIN3; 799 women had Normal/CIN1 histology; 46 women had undetermined disease status. The results of the APTIMA HPV Assay by the consensus histology review panel diagnosis are presented in the table below. NILM ≥ 30 Years Population: Results of the APTIMA HPV Assay and Consensus Histology Review Panel Diagnosis | APTIMA HPV Assay Result* | HPV DNA Test | Consensus Histology Review Panel Diagnosis | | | | | | | | --- | --- | --- | --- | --- | --- | --- | --- | --- | | | | Undetermined | Normal | CIN1 | CIN2 | CIN3 | Cancer | Total | | Positive | Positive | 11 | 212 | 11 | 4 | 7 | 2 | 247 | | Positive | Negative | 7 | 59 | 0 | 1 | 0 | 1 | 68 | | Positive | No Result** | 3 | 16 | 1 | 0 | 0 | 0 | 20 | | Negative | Positive | 10 | 170 | 8 | 2 | 1 | 0 | 191 | | Negative | Negative | 15 | 313 | 9 | 1 | 0 | 0 | 338 | | Negative | No Result** | 0 | 0 | 0 | 1 | 0 | 0 | 1 | | Total | | 46 | 770 | 29 | 9 | 8 | 3*** | 865 | *All samples had final valid results (upon initial testing or after resolution of initial invalids per procedure). **21 women with APTIMA HPV Assay results did not have HPV DNA test results primarily due to insufficient volume of the cytology specimen. ***Three women had adenocarcinoma in situ (AIS). In total, 10,052 women had unverified (including undetermined) disease status. Because only randomly selected women with negative results for both the APTIMA HPV Assay and the FDA-approved HPV DNA test were referred to colposcopy, the proportion of women with unverified disease status was high in this group (96.6%). To adjust for this verification bias, a multiple imputation method was used to estimate the number of women with disease that would have been identified if all women had undergone colposcopy. Both verification-bias adjusted performance estimates and unadjusted performance estimates based on the 819 women with verified disease status are presented below. PMA P100042: FDA Summary of Safety and Effectiveness Data page 27 {27} NILM ≥ 30 Years Population: Classification of Evaluable NILM Women by APTIMA HPV Assay and HPV DNA Test Results, Disease Status (≥CIN2 and ≥CIN3), and Disease Verification Status | APTIMA HPV Assay Result* | HPV DNA Test | Total Women | Verified Disease Status: ≥CIN2 | | Verified Disease Status: ≥CIN3 | | Unverified Disease Status | | --- | --- | --- | --- | --- | --- | --- | --- | | | | | Diseased Women (≥CIN2) | Non-Diseased Women ( PMA P100042: FDA Summary of Safety and Effectiveness Data page 34 {34} For the composite comparator analysis, samples were classified positive if the HPV DNA test and the E6/E7 RT-PCR sequencing test were both positive; negative if the HPV DNA test and the E6/E7 RT-PCR sequencing test were both negative; and indeterminate if the tests were discordant, or if one or both tests returned an invalid or indeterminate result. Positive and negative percent agreements and associated 95% confidence intervals were calculated. Indeterminate results were not included in the agreement calculations. Results are presented for the ASC-US population (≥ 21 years) and the NILM population (≥ 30 years) in the tables below. **ASC-US ≥ 21 Years Population: APTIMA HPV Assay Agreement Results with a Composite Comparator (n=217)** | | Composite Comparator | | | Total | | | --- | --- | --- | --- | --- | --- | | | | Positive | Negative | | Indeterminate | | APTIMA HPV Assay | Positive | 89 | 0 | 27 | 116 | | | Negative | 2 | 86 | 13 | 101 | | | Total | 91 | 86 |… --- **Source:** [https://fda.innolitics.com/device/P100042](https://fda.innolitics.com/device/P100042) **Published by [Innolitics](https://innolitics.com)** — a medical-device software consultancy. We help companies design, build, and clear FDA-regulated software and AI/ML devices. 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