Aptima Combo 2 Assay (Panther System), Aptima Combo 2 Assay (Tigris) System)

K200866 · Hologic, Inc. · QEP · May 17, 2020 · Microbiology

Device Facts

• Record ID: K200866
• Device Name: Aptima Combo 2 Assay (Panther System), Aptima Combo 2 Assay (Tigris) System)
• Applicant: Hologic, Inc.
• Product Code: QEP · Microbiology
• Decision Date: May 17, 2020
• Decision: SESE
• Submission Type: Special
• Regulation: 21 CFR 866.3393
• Device Class: Class 2

Intended Use

The Aptima Combo 2 Assay is a target amplification nucleic acid probe test that utilizes target capture for the in vitro qualitative detection and differentiation of ribosomal RNA (rRNA) from Chlamydia trachomatis (CT) and/or Neisseria gonorrhoeae (GC) to aid in the diagnosis of chlamydial and/or gonococcal disease using the Panther System as specified. On the Panther System, the assay may be used to test the following specimens from symptomatic and asymptomatic individuals: clinician-collected endocervical, PreservCyt® Solution liquid Pap specimens, vaginal, throat, rectal, and male urethral swab specimens; patient-collected vaginal swab specimens1, and female and male urine specimens. The Aptima Combo 20 Assay is a target amplification nucleic acid probe test that utilizes target capture for the in vitro qualitative detection and differentiation of ribosomal RNA (rRNA) from Chlamydia trachomatis (CT) and/or Neisseria gonorrhoeae (GC) to aid in the diagnosis of chlamydial and/or gonococcal urogenital disease using the Tigris® DTS® Automated Analyzer or semi-automated instrumentation as specified. The assay may be used to test the following specimens from symptomatic individuals: clinician-collected endocervical, vaginal and male urethral swab specimens; and female and male urine specimens. The assay may be used to test the following specimens from asymptomatic individuals: clinician-collected endocervical, vaginal and male urethral swab specimens; patient-collected vaginal swab specimens'; and female and male urine specimens. The assay is also intended for use with the testing of gynecological specimens, from both symptomatic and asymptomatic patients, collected in the PreservCyt® Solution.

Device Story

The Aptima Combo 2 Assay is an in vitro diagnostic test for Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (GC). It processes clinical specimens (swabs, urine, PreservCyt) using target capture with magnetic microparticles to isolate rRNA. The system performs Transcription-Mediated Amplification (TMA) to replicate specific rRNA regions. Detection uses Hybridization Protection Assay (HPA) with acridinium ester-labeled chemiluminescent probes. The updated assay incorporates a tandem probe for CT to detect emerging variants. The system measures light emission (Relative Light Units) in a luminometer; kinetic profiles (flasher for CT, glower for GC) differentiate the targets. Used in clinical laboratories, the assay is fully automated on Panther or Tigris systems. Results are interpreted by the system based on RLU cut-offs and kinetic curves. The output assists clinicians in diagnosing STIs, enabling targeted patient treatment.

Clinical Evidence

Bench testing only. Verification and validation activities were conducted based on risk analysis to confirm the performance of the modified dual-probe system compared to the original single-probe design.

Technological Characteristics

Nucleic acid amplification test using target capture, Transcription-Mediated Amplification (TMA), and Hybridization Protection Assay (HPA). Employs magnetic microparticles for target isolation. Detection via acridinium ester-labeled chemiluminescent probes with kinetic differentiation (flasher/glower). Automated on Panther/Tigris systems. Reformulated probe reagent includes dual/tandem CT probes for variant coverage.

Indications for Use

Indicated for symptomatic and asymptomatic individuals for the qualitative detection and differentiation of Chlamydia trachomatis and Neisseria gonorrhoeae rRNA to aid in the diagnosis of chlamydial and gonococcal disease.

Regulatory Classification

Identification

A device to detect nucleic acids from non-viral microorganism(s) causing sexually transmitted infections and associated resistance marker(s) is an in vitro diagnostic device intended for the detection and identification of nucleic acids from non-viral microorganism(s) and their associated resistance markers in clinical specimens collected from patients suspected of sexually transmitted infections. The device is intended to aid in the diagnosis of non-viral sexually transmitted infections in conjunction with other clinical and laboratory data. These devices do not provide confirmation of antibiotic susceptibility since mechanisms of resistance may exist that are not detected by the device.

Special Controls

A device to detect nucleic acids from non-viral microorganism(s) causing sexually transmitted infections and associated resistance marker(s) must comply with the following special controls: (1) The intended use for the 21 CFR 809.10 labeling must include a detailed description of targets the device detects, the results provided to the user, the clinical indications appropriate for test use, and the specific population(s) for which the device is intended. (2) Any sample collection device used must be FDA-cleared, -approved, or -classified as 510(k) exempt (standalone or as part of a test system) for the collection of specimen types claimed by this device: alternatively, the sample collection device must be cleared in a premarket submission as a part of this device. (3) The 21 CFR 809.10(b) labeling must include: (i) A detailed device description, including reagents, instruments, ancillary materials, all control elements, and a detailed explanation of the methodology, including all pre-analytical methods for processing of specimens; (ii) Detailed discussion of the performance characteristics of the device for all claimed specimen types based on analytical studies, including, but not limited to. Limit of Detection, inclusivity, cross-reactivity, interfering substances, competitive inhibition, carryover/cross contamination, specimen stability, with-in lab precision, and reproducibility, as appropriate; (iii) Detailed descriptions of the test procedure, the interpretation of test results for clinical specimens, and acceptance criteria for any quality control testing. (iv) Limiting statements indicating that: (A)a negative test result does not preclude the possibility of infection; (B) the test results should be interpreted in conjunction with other clinical and laboratory data available to the clinician; (C) reliable results are dependent on adequate specimen collection, transport, storage, and processing. Failure to observe procedures in any one of these steps can lead to incorrect results; and (D)if appropriate (e.g., recommended by CDC, by current well-accepted clinical guidelines, or by published peer reviewed research), that the clinical performance is inferior in a specific clinical subpopulation or for a specific claimed specimen type. (v) If the device is intended to detect antimicrobial resistance markers, limiting statements, as appropriate, indicating that: (A)negative results for claimed resistance markers do not indicate susceptibility of detected microorganisms, as resistance markers not measured by the assay or other potential mechanisms of antibiotic resistance may be present; (B) detection of resistance markers cannot be definitively linked to specific microorganisms and the source of a detected resistance marker may be an organism not detected by the assay, including colonizing flora; (C) detection of antibiotic resistance markers may not correlate with phenotypic gene expression; and (D) therapeutic failure or success cannot be determined based on the assay results, since nucleic acid may persist following appropriate antimicrobial therapy. (4) Design verification and validation must include: (i) Detailed device description documentation, including, but not limited to, methodology from obtaining sample to result, design of primer/probe sequences, rationale for target sequence selection, and computational path from collected raw data to reported result (e.g., how collected raw signals are converted into a reported result). (ii) Detailed documentation of analytical studies including but not limited to, Limit of Detection, inclusivity, cross-reactivity, microbial interference, interfering substances, competitive inhibition, carryover/cross contamination, specimen stability, with-in lab precision, and reproducibility, as appropriate. (iii) Detailed documentation and performance results from a clinical study that includes prospective (sequential) samples for each claimed specimen type and, when determined to be appropriate by FDA, additional characterized clinical samples. The study must be performed on a study population consistent with the intended use population and compare the device performance to results obtained from FDA accepted comparator methods. Documentation from the clinical studies must include the clinical study protocol (including a predefined statistical analysis plan) study report, testing results, and results of all statistical analyses. (iv) A detailed description of the impact of any software, including, but not limited to, software applications and hardware-based devices that incorporate software, on the device's functions.

*Classification.* Class II (special controls). The special controls for this device are:(1) The intended use for the labeling required under § 809.10 of this chapter must include a detailed description of targets the device detects, the results provided to the user, the clinical indications appropriate for test use, and the specific population(s) for which the device is intended. (2) Any sample collection device used must be FDA-cleared, -approved, or -classified as 510(k) exempt (standalone or as part of a test system) for the collection of specimen types claimed by this device; alternatively, the sample collection device must be cleared in a premarket submission as a part of this device. (3) The labeling required under § 809.10(b) of this chapter must include: (i) A detailed device description, including reagents, instruments, ancillary materials, all control elements, and a detailed explanation of the methodology, including all pre-analytical methods for processing of specimens; (ii) Detailed discussion of the performance characteristics of the device for all claimed specimen types based on analytical studies, including Limit of Detection, inclusivity, cross-reactivity, interfering substances, competitive inhibition, carryover/cross contamination, specimen stability, within lab precision, and reproducibility, as appropriate; (iii) Detailed descriptions of the test procedure, the interpretation of test results for clinical specimens, and acceptance criteria for any quality control testing; (iv) Limiting statements indicating that: (A) A negative test result does not preclude the possibility of infection; (B) The test results should be interpreted in conjunction with other clinical and laboratory data available to the clinician; (C) Reliable results are dependent on adequate specimen collection, transport, storage, and processing. Failure to observe proper procedures in any one of these steps can lead to incorrect results; and (D) If appropriate ( *e.g.,* recommended by the Centers for Disease Control and Prevention, by current well-accepted clinical guidelines, or by published peer reviewed research), that the clinical performance is inferior in a specific clinical subpopulation or for a specific claimed specimen type; and(v) If the device is intended to detect antimicrobial resistance markers, limiting statements, as appropriate, indicating that: (A) Negative results for claimed resistance markers do not indicate susceptibility of detected microorganisms, as resistance markers not measured by the assay or other potential mechanisms of antibiotic resistance may be present; (B) Detection of resistance markers cannot be definitively linked to specific microorganisms and the source of a detected resistance marker may be an organism not detected by the assay, including colonizing flora; (C) Detection of antibiotic resistance markers may not correlate with phenotypic gene expression; and (D) Therapeutic failure or success cannot be determined based on the assay results, since nucleic acid may persist following appropriate antimicrobial therapy. (4) Design verification and validation must include: (i) Detailed device description documentation, including methodology from obtaining sample to result, design of primer/probe sequences, rationale for target sequence selection, and computational path from collected raw data to reported result ( *e.g.,* how collected raw signals are converted into a reported result).(ii) Detailed documentation of analytical studies, including, Limit of Detection, inclusivity, cross-reactivity, microbial interference, interfering substances, competitive inhibition, carryover/cross contamination, specimen stability, within lab precision, and reproducibility, as appropriate. (iii) Detailed documentation and performance results from a clinical study that includes prospective (sequential) samples for each claimed specimen type and, when determined to be appropriate by FDA, additional characterized clinical samples. The study must be performed on a study population consistent with the intended use population and compare the device performance to results obtained from FDA accepted comparator methods. Documentation from the clinical studies must include the clinical study protocol (including a predefined statistical analysis plan) study report, testing results, and results of all statistical analyses. (iv) A detailed description of the impact of any software, including software applications and hardware-based devices that incorporate software, on the device's functions.

Predicate Devices

• Aptima Combo 2 Assay on Panther and Tigris Systems (K200436)

Submission Summary (Full Text)

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Image /page/0/Picture/0 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). The logo consists of two parts: the symbol of the Department of Health & Human Services on the left and the FDA acronym along with the full name of the agency on the right. The FDA part of the logo is in blue, with the acronym in a square and the full name written out next to it.

May 17, 2020

Hologic, Inc. Anila Tarte Regulatory Affairs Specialist 10210 Genetic Center Drive San Diego, California 92121

#### Re: K200866

Trade/Device Name: Aptima Combo 2 Assay (Panther System) and Aptima Combo 2 Assay (Tigris System) Regulation Number: 21 CFR 866.3393 Regulation Name: Nucleic Acid Detection System for Non-Viral Microorganism(s) Causing Sexually Transmitted Infections. Regulatory Class: Class II Product Code: QEP, LSL, MKZ Dated: March 30, 2020 Received: April 1, 2020

Dear Anila Tarte:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

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Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or post-marketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-products); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.

For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email ([e-mail]) or phone [phone] or 301-796-7100).

Sincerely.

Steven Gitterman, M.D., Ph.D. Deputy Director Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics and Radiological Health Office of Product Evaluation and Quality Center for Devices and Radiological Health

Enclosure

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# HOLOGIC®

# 510(k) SUMMARY

# Aptima Combo 2® Assay (Panther® and Tigris® DTS® System)

## I. SUBMITTER

Hologic, Inc. 10210 Genetic Center Drive San Diego, CA 92121

## Contact Information:

| | Anila Tarte | |
|--------|-------------------------------|--|
| | Regulatory Affairs Specialist | |
| Phone: | 858-410-8055 | |
| Email: | [e-mail] | |

Date Prepared: May 14, 2020

## II. DEVICES

| Proprietary Name: | Aptima Combo 2® Assay (Panther® System) |
|--------------------------|---------------------------------------------------------------------------------------------------------|
| Classification Name: | Nucleic Acid Detection System for Non-Viral Microorganism(s)<br>Causing Sexually Transmitted Infections |
| Regulation Number: | 866.3393 |
| Regulatory Class: | Class II |
| Product Code: | QEP |
| Subsequent Product Code: | MKZ, LSL |
| Proprietary Name: | Aptima Combo 2® Assay (Tigris® DTS® System) |
| Classification Name: | Nucleic Acid Detection System for Non-Viral Microorganism(s)<br>Causing Sexually Transmitted Infections |
| Regulation Number: | 866.3393 |
| Regulatory Class: | Class II |
| Product Code: | OEP |

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#### III. PREDICATE DEVICE

The predicate device is the Aptima Combo 2 Assay on Panther and Tigris Systems (K200436; cleared 03/24/2020). The predicate device has not been subject to a design-related recall.

#### IV. DEVICE DESCRIPTIONS

The Aptima Combo 2 Assay combines the technologies of target capture, TMA, and DKA. Specimens are collected and transferred into their respective specimen transport tubes. The transport solutions in these tubes release the rRNA targets and protect them from degradation during storage. When the Aptima Combo 2 Assay is performed in the laboratory, the target rRNA molecules are isolated from specimens by use of capture oligomers via target capture that utilizes magnetic microparticles. The capture oligomers contain sequences complementary to specific regions of the target molecules as well as a string of deoxyadenosine residues. A separate capture oligomer is used for each target. During the hybridization step, the sequence specific regions of the capture oligomers bind to specific regions of the target molecules. 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 target molecules bound to them, are pulled to the side of the reaction vessel using magnets and the supernatant is aspirated. The particles are washed to remove residual specimen matrix that may contain amplification reaction inhibitors. After the target capture steps are completed, the specimens are ready for amplification.

Target amplification assays are based on the ability of complementary oligonucleotide primers to specifically anneal and allow enzymatic amplification of the target nucleic acid strands. The Aptima Combo 2 Assay replicates a specific region of the 23S rRNA from CT and a specific region of the 16S rRNA from GC via DNA intermediates. A unique set of primers is used for each target molecule. Detection of the rRNA amplification product sequences (amplicon) is achieved using nucleic acid hybridization. Single-stranded nucleic acid chemiluminescent probes, which are complementary to a region of each target amplicon, are labeled with different acridinium ester molecules. The updated version of the Aptima Combo 2 assay incorporates a

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second CT probe, complementary to a unique region of the existing CT amplicon. This tandem probe provides detection coverage for the variant strains of C. trachomatis that emerged in 2019. The labeled probes combine with amplicon to form stable hybrids. The Selection Reagent differentiates hybridized from unhybridized probe, eliminating the generation of signal from unhybridized probe. During the detection step, light emitted from the labeled hybrids is measured as photon signals in a luminometer, and are reported as Relative Light Units (RLU). In DKA, differences in the kinetic profiles of the CT and GC labeled probes allow for the differentiation of signal; kinetic profiles are derived from measurements of photon output during the detection read time. The chemiluminescent detection for CT signal has very rapid kinetics and has the "flasher" kinetic type. The chemiluminescent detection reaction for GC signal is relatively slower and has the "glower" kinetic type. Assay results are determined by a cut-off based on the total RLU and the kinetic curve type.

## V. DESCRIPTION OF DEVICE MODIFICATION

The clearance of this Special 510(k) application supports a change in formulation to the Probe reagent contained in the Aptima Combo 2 assay. Reformulation of the Probe reagent was necessary to detect recently emerged variants of Chlamydia trachomatis (CT) that were discovered outside of the U.S. using the Panther System or Tigris System. The updated version of the Aptima Combo 2 Assay (termed "updated AC2 assay") includes dual (redundant) CT detection probe, which not only identifies all recent variants of CT, but is also intended to provide diagnostic protection against future genetic variants within the AC2 probe region.

| Table 1: Aptima Combo 2 Assay - Catalog Numbers | |
|-------------------------------------------------|--|
|-------------------------------------------------|--|

| Kit Description | Current Kit<br>Cat. No. | Updated Kit<br>Cat. No. |
|-------------------------------------------------------|-------------------------|-------------------------|
| Aptima Combo 2 Assay, 100-Test Kit (Panther System) | 302923 | PRD-05576 |
| Aptima Combo 2 Assay, 250-Test Kit (Panther System) | 303094 | PRD-05571 |
| Aptima Combo 2 Assay, 250-Test Kit (Tigris System) | 301130 | PRD-05572 |
| Aptima Combo 2 Assay, 1000-Test Kit (Tigris System) * | 301130B | PRD-05572B |

* AC2 for Tigris 1000-Test Kit is packaged to contain four 250-Test Kits

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#### VI. INDICATIONS FOR USE

#### Intended Use - Aptima Combo 2 Assay (Panther)

The Aptima Combo 2 Assay is a target amplification nucleic acid probe test that utilizes target capture for the in vitro qualitative detection and differentiation of ribosomal RNA (rRNA) from Chlamydia trachomatis (CT) and/or Neisseria gonorrhoeae (GC) to aid in the diagnosis of chlamydial and/or gonococcal disease using the Panther System as specified. On the Panther System, the assay may be used to test the following specimens from symptomatic and asymptomatic individuals: clinician-collected endocervical, PreservCyt® Solution liquid Pap specimens, vaginal, throat, rectal, and male urethral swab specimens; patient-collected vaginal swab specimens1, and female and male urine specimens.

'Patient-collected vaginal swab specimens are an option for screening women when a pelvic exam is not otherwise indicated. The Aptima Multitest Swab Specimen Collection Kit has not been evaluated for home use.

#### Intended Use - Aptima Combo 2 Assay (Tigris)

The Aptima Combo 20 Assay is a target amplification nucleic acid probe test that utilizes target capture for the in vitro qualitative detection and differentiation of ribosomal RNA (rRNA) from Chlamydia trachomatis (CT) and/or Neisseria gonorrhoeae (GC) to aid in the diagnosis of chlamydial and/or gonococcal urogenital disease using the Tigris® DTS® Automated Analyzer or semi-automated instrumentation as specified. The assay may be used to test the following specimens from symptomatic individuals: clinician-collected endocervical, vaginal and male urethral swab specimens; and female and male urine specimens. The assay may be used to test the following specimens from asymptomatic individuals: clinician-collected endocervical, vaginal and male urethral swab specimens; patient-collected vaginal swab specimens'; and female and male urine specimens. The assay is also intended for use with the testing of gynecological specimens, from both symptomatic and asymptomatic patients, collected in the PreservCyt® Solution.

IPatient-collected vaginal swab specimens are an option for screening women when a pelvic exam is not otherwise indicated. The Aptima Multitest Swab Specimen Collection Kit is not for home use.

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# VII. COMPARISON OF TECHNOLOGICAL CHARACTERISTICS WITH THE PREDICATE DEVICES

A comparison of the subject device to the predicate devices is summarized in Table 2 (AC2 assay on Panther) and Table 3 (AC2 assay on Tigris). Use of the updated AC2 assay does not change the principles of procedure, intended use, or primary technological characteristics. The similarities and differences between the subject and predicate devices are further discussed following the substantial equivalence tables. This discussion is the same for each assay.

Table 2: Comparison Between Predicate Device and Subject Device - AC2 Assay on the Panther System

| Item | Predicate Device<br>AC2 Assay (Panther)<br>K200436 | Subject Device AC2Assay (Panther)<br>K200866 |
|-----------------------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Technology<br>Principle of<br>Operation | Target Capture (TC), Transcription-Mediated<br>Amplification (TMA), Hybridization<br>Protection Assay (HPA) | Same |
| Platform | Automated Panther System | Same |
| Function | Detection and differentiation of rRNA from<br>Chlamydia trachomatis and Neisseria<br>gonorrhoeae | Same |
| Organisms<br>Detected | Chlamydia trachomatis (CT) and/or<br>Neisseria gonorrhoeae (GC) | Same |
| Patient<br>Population | Symptomatic and asymptomatic individuals | Same |
| Intended Use | The Aptima Combo 2 Assay is a target<br>amplification nucleic acid probe test that<br>utilizes target capture for the in vitro<br>qualitative detection and differentiation of<br>ribosomal RNA (rRNA) from Chlamydia<br>trachomatis (CT) and/or Neisseria<br>gonorrhoeae (GC) to aid in the diagnosis of<br>chlamydial and/or gonococcal urogenital<br>disease using the Panther® System as<br>specified On the Panther System, the assay<br>may be used to test the following specimens<br>from<br>symptomatic and asymptomatic individuals:<br>clinician-collected endocervical, vaginal,<br>throat, rectal, and male urethral swab<br>specimens clinician-collected gynecological | The Aptima Combo 2® Assay is a target<br>amplification nucleic acid probe test that<br>utilizes target capture for the in vitro<br>qualitative detection and differentiation<br>of ribosomal RNA (rRNA) from<br>Chlamydia trachomatis (CT) and/or<br>Neisseria gonorrhoeae (GC) to aid in<br>the diagnosis of chlamydial and/or<br>gonococcal disease using the Panther®<br>System as specified. On the Panther<br>System, the assay may be used to test<br>the following specimens from<br>symptomatic and asymptomatic<br>individuals: clinician-collected<br>endocervical, PreservCyt® Solution<br>liquid pan specimens vaginal throat |
| Same* | | |

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| Item | Predicate Device<br>AC2 Assay (Panther)<br>K200436 | Subject Device AC2Assay (Panther)<br>K200866 |
|------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| | specimens collected in the PreservCyt®<br>Solution, patient-collected vaginal swab<br>specimens,1 and female and male urine<br>specimens. | rectal, and male urethral swab<br>specimens, patient-collected vaginal<br>swab specimens,1 and female and male<br>urine specimens. |
| | 1Patient-collected vaginal swab specimens are an<br>option for screening women when a pelvic exam<br>is not otherwise indicated. The Aptima Multitest<br>Swab Specimen Collection Kits have not been<br>evaluated for home use. | 1Patient-collected vaginal swab specimens<br>are an option for screening women when a<br>pelvic exam is not otherwise indicated. The<br>Aptima Multitest Swab Specimen Collection<br>Kit has not been evaluated for home use. |

* Edits to the intended use were made for clarification purposes only. Specimen types were aligned and/or grouped into respective clinician-collected and patient-collected specimen types

| | Table 3: Comparison Between Predicate Device and Subject Device - AC2 Assay |
|----------------------|-----------------------------------------------------------------------------|
| on the Tigris System | |

| Item | Predicate Device<br>AC2 Assay (Tigris)<br>K200436 | Subject Device<br>AC2 Assay (Tigris)<br>K200866 |
|-----------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------|
| Technology<br>Principle of<br>Operation | Target Capture (TC), Transcription-Mediated<br>Amplification (TMA), Hybridization Protection Assay<br>(HPA) | Same |
| Platform | Automated Tigris System | Same |
| Function | Detection and differentiation of rRNA from Chlamydia<br>trachomatis and Neisseria gonorrhoeae | Same |
| Organisms<br>Detected | Chlamydia trachomatis (CT) and/or Neisseria<br>gonorrhoeae (GC) | Same |
| Patient<br>Population | Symptomatic and asymptomatic individuals | Same |
| Intended Use | The Aptima Combo 2 Assay is a target amplification<br>nucleic acid probe test that utilizes target capture for the in<br>vitro qualitative detection and differentiation of ribosomal<br>RNA (rRNA) from Chlamydia trachomatis (CT) and/or<br>Neisseria gonorrhoeae (GC) to aid in the diagnosis of<br>chlamydial and/or gonococcal urogenital disease using the<br>Tigris® DTS® Automated Analyzer. On the Tigris DTS<br>system, the assay may be used to test the following<br>specimens from symptomatic individuals: clinician-<br>collected endocervical, vaginal and male urethral swab<br>specimens; and female and male urine specimens. The<br>assay may be used to test the following specimens from<br>asymptomatic individuals: clinician-collected<br>endocervical, vaginal and male urethral swab specimens;<br>patient-collected vaginal swab specimens1; and female and | Same |

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| Item | Predicate Device<br>AC2 Assay (Tigris)<br>K200436 | Subject Device<br>AC2 Assay (Tigris)<br>K200866 |
|------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-------------------------------------------------|
| | male urine specimens. The assay is also intended for use<br>with the testing of gynecological specimens, from both<br>symptomatic and asymptomatic patients, collected in the<br>PreservCyt® Solution. | |
| | 1Patient-collected vaginal swab specimens are an option for screening<br>women when a pelvic exam is not otherwise indicated. The vaginal and<br>multitest swab specimen collection kits are not for home use. | |

#### Similarities

Both the predicate and subject devices utilize the same technology and principles of operation, mechanisms of action, and run on the same automated instrument systems. There are no changes to the assay kit configuration, intended use, results interpretation, or existing performance of the assay. Additionally, the proposed changes do not affect the existing Aptima Controls Kit, Aptima ancillary or collection kits, or the software and hardware associated with the use of the Panther or Tigris systems.

#### Differences

Changes to the user interface are minimal and include updated packaging and labeling to differentiate between the current AC2 assay kit and the updated AC2 assay kit. Package insert changes include new kit catalog numbers and updates to the Analytical Performance section demonstrating detection of all recent CT variants using the updated AC2 assay.

## VIII. Design Control Activities

Hologic's overall product development activities are conducted per procedure, 'Product Development Procedure' which is in conformance with the design control requirements as specified in 21 CFR 820.30. Verification testing was performed to confirm clinical comparability between the current Probe and reformulated Probe reagents. The completed verification studies demonstrate that the reformulated Probe reagent does not impact assay performance, assay safety and effectiveness, and confirms that the modified assay meets the design input requirements.

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Hologic risk analysis activities are conducted per Product Safety Risk Management Procedure which is in conformance with ISO 14971:2007. Based on the results of the risk analysis and verification activities, and in accordance with ISO 14971:2007, all risks are reduced as far as possible and meet the pre-defined acceptability criteria. There were no hazards that fell within the "Undesirable" or "Unacceptable" residual risk regions. The device modifications do not introduce any new hazards or increase the overall residual risk as compared to the currently marketed products.

#### IX. ASSAY PERFORMANCE

Performance of the updated AC2 Assay was evaluated. The result of this evaluation demonstrated that existing performance and claims of the assay were not impacted due to the reformulated Probe reagent.

#### Brief Description of Non-Clinical Data

The following analytical (non-clinical) studies were conducted to support the clearance of the updated AC2 assay on the Panther System and Tigris System.

#### Limit of Detection – Panther and Tigris

The analytical sensitivity for the Finnish variant of Chlamydia trachomatis (FI-nvCT) was determined by testing dilutions of in vitro transcripts in negative urine specimens, negative ThinPrep specimens, and simulated swab matrix specimens. Thirty replicates of each dilution were tested on both the Panther system and Tigris system with each of three reagent lots of the updated AC2 assay for a total of 90 replicates per specimen type. The analytical sensitivity was determined to be less than one IFU per assay in urine, ThinPrep, and simulated swab matrix specimens. The detection capabilities of the updated version of the AC2 assay were confirmed across multiple CT variants.

#### Clinical Comparability

The clinical specimen agreement between the current version and updated version of the AC2 assay was evaluated using remnant swab specimens collected from patients undergoing CT

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and/or GC screening. A single replicate of each specimen was tested with both the current version and the updated version of the AC2 assay on the Panther System.

Table 4 and Table 5 show clinical comparison results for the CT and GC positive and negative percent agreement, respectively. The overall agreement was >99.0% for both CT and GC.

| | | Current AC2 Assay | |
|--------------------------------------------------------------|-------------|-------------------|-------------|
| | | CT Positive | CT Negative |
| Updated AC2<br>Assay | CT Positive | 49 | 3 |
| | CT Negative | 0 | 273 |
| Positive Percent Agreement (95% C.I.): 100% (92.7% - 100%) | | | |
| Negative Percent Agreement (95% C.I.): 98.9% (96.9% - 99.6%) | | | |

#### Table 4: Clinical Specimen Comparison Results - CT

#### Table 5: Clinical Specimen Comparison Results - GC

| | | Current AC2 Assay | |
|--------------------------------------------------------------|-------------|-------------------|-------------|
| | | GC Positive | GC Negative |
| Updated AC2<br>Assay | GC Positive | 47 | 1 |
| | GC Negative | 0 | 275 |
| Positive Percent Agreement (95% C.I.): 100% (92.4% - 100%) | | | |
| Negative Percent Agreement (95% C.I.): 99.6% (98.0% - 99.9%) | | | |

#### CT/GC Clinical Sample Agreement

The clinical panel agreement study evaluated the equivalence between the current and updated versions of the AC2 assay using 20 prepared CT/GC clinical panels containing 0 to 2,500 IFU/mL of wild type CT, 0 to 500 IFU/mL of FI-nvCT, and 0 to 125,000 CFU/mL of GC in urine specimens. Each of the 20 panels were tested in triplicate in two runs per day, on three Panther systems, by two operators, using three lots of reagents over seven days.

The results show 100% (97.6-100%) total CT and GC agreement to the expected panel result for the updated AC2 assay. Results also show 100% (97.6-100%) total CT and GC agreement to the expected panel result for the current AC2 assay, with the exception of the moderate (0.2 IFU/mL) FI-nvCT only panel, which had 98.2% (93.5-99.5%) CT agreement and 99.1% (94.9-99.8%) GC agreement. The percent agreement to the expected result for the detection of wild

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type CT and GC is comparable between the current AC2 assay and the updated AC2 assay. In addition, the observed variability of the current AC2 and the updated AC2 assays was comparable between instruments, lots, operators, days, and runs.

## Microorganism Cross-Reactivity and Microbial Interference

The analytical specificity and microbial interference of the updated version of the AC2 assay was evaluated using 86 microorganisms consisting primarily of viral, bacterial, and yeast strains. Each pool of microorganisms was tested with and without the presence of FI-nvCT in vitro transcripts at a concentration of 3x LoD. None of the microorganisms tested were found to have an impact on the detection capabilities or analytical specificity of the updated version of the AC2 assay.

| Pool ID | Microorganism | Pool ID | Microorganism |
|---------|----------------------------------------------------------------------------------------------------------------------|---------|---------------------------------------------------------------------------------------------------------------|
| 0 | SVSM (Control) | | |
| 1 | Acinetobacter lwoffii<br>Actinomyces israelii<br>Alcaligenes faecalis<br>Anaerococcus vaginalis | 12 | Proteus vulgaris<br>Shigella dysenteriae<br>Shigella flexneri<br>Shigella sonneri |
| 2 | Arcanobacterium haemolyticum<br>Atopobium vaginae<br>Bacteroides fragilis<br>Bacteroides oralis | 13 | Stenotrophomonas maltophilia<br>Streptococcus agalactiae<br>Streptococcus anginosus<br>Streptococcus pyogenes |
| 3 | Bifidobacterium adolescentis<br>Bordatella parapertussis<br>Campylobacter jejuni<br>Campylobacter rectus | 14 | Ureaplasma parvum<br>Ureaplasma urealyticum<br>Veillonella parvula<br>Burkholderia cepacia |
| 4 | Citrobacter koseri<br>Corynebacterium diptheria<br>Corynebacterium genitalium<br>Corynebacterium pseudodiptheriticum | 15 | Clostridium difficile<br>Prevotella bivia<br>Candida albicans<br>Cryptococcus neoformans |
| 5 | Eggerthella lenta<br>Enterobacter cloacae<br>Enterococcus faecalis<br>Escherichia coli | 16 | Entamoeba histolytica<br>Giardia lamblia<br>Pentatrichomonas hominis<br>Trichomonas vaginalis |

Table 6: Cross-Reactivity Microorganisms

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| Pool ID | Microorganism | Pool ID | Microorganism |
|---------|--------------------------------------------------------------------------------------------------------------|---------|-------------------------------------------------------------------------------------|
| 6 | Fusobacterium necrophorum<br>Fusobacterium nucleatum<br>Gardnerella vaginalis<br>Helicobacter pylori | 17 | Adenovirus Type 07A<br>Coronavirus 229E<br>Coxsackievirus B3<br>Echovirus Type 11 |
| 7 | Haemophilus ducreyi<br>Haemophilus parahaemolyticus<br>Haemophilus parainfluenzae<br>Klebsiella pneumoniae | 18 | Enterovirus Type 68<br>Epstein-Barr virus<br>Hepatitis B Virus<br>Hepatitis C Virus |
| 8 | Lactobacillus acidophilus<br>Legionella (Tatlockia) micdadei<br>Legionella jordanis<br>Leptotrichia buccalis | 19 | HIV<br>HPV 16 (SiHa cells)<br>HPV 18 (HeLa cells)<br>Human Metapneumovirus Type 20 |
| 9 | Listeria monocytogenes<br>Megasphaera Type 1<br>Mobiluncus curtisii<br>Moraxella catarrhalis | 20 | HSV I<br>HSV II<br>Influenza A H3N2<br>Influenza B Massachusetts/2/12 |
| 10a | Mycoplasma genitalium<br>Mycoplasma hominis<br>Mycoplasma pneumoniae | 21 | Norovirus Group II<br>Respiratory Syncytial virus Type B<br>Rhinovirus A16 |
| 10b | Neisseria gonorrhoeae | | |
| 11 | Peptostreptococcus micros<br>Propionibacterium acnes<br>Staphylococcus aureus<br>Staphylococcus epidermidis | 22 | Chlamydia pneumoniae<br>Chlamydia psittaci<br>Chlamydia psittaci |

# X. CONCLUSIONS

A comparison of the intended use, technological characteristics, and results from the analytical performance studies demonstrate that the updated AC2 assay on the Panther and Tigris systems performs comparably to the predicate device.