ARCHITECT CA 19-9XR ASSAY
Device Facts
| Record ID | K052000 |
|---|---|
| Device Name | ARCHITECT CA 19-9XR ASSAY |
| Applicant | Fujirebio Diagnostics,Inc. |
| Product Code | NIG · Immunology |
| Decision Date | Oct 25, 2005 |
| Decision | SESE |
| Submission Type | Traditional |
| Regulation | 21 CFR 866.6010 |
| Device Class | Class 2 |
Indications for Use
The ARCHITECT® CA 19-9™_XR assay is a Chemiluminescent Microparticle Immunoassay (CMIA) for the quantitative determination of 1116-NS-19-9 reactive determinants inhuman serum and plasma on the ARCHITECT i System. The ARCHITECT® CA 19-9™_XR assay is to be used as an aid in the management of pancreatic cancer patients with a detectable level of Ca 19-9 at some point in their disease process and in conjunction with other clinical methods. The ARCHITECT® CA 19-9™_XR Calibrators are for the calibration of the ARCHITECT i System when used for the quantitative determination of 1116-19-9 reactive determinants in human serum and plasma. Refer to the ARCHITECT® CA 19-9™ₓₗ reagent package insert for additional information. The ARCHITECT® CA 19-9™ₓₗ Controls are for the estimation of test precision and the detection of systematic analytical deviations of the ARCHITECT i System (reagents, calibrators, and instrument), when used for the quantitative determination of 1116-NS-19-9 reactive determinants in human serum and plasma. Refer to the ARCHITECT® CA 19-9™ₓₗ reagent package insert for additional information.
Device Story
The ARCHITECT CA 19-9xR assay is a two-step chemiluminescent microparticle immunoassay (CMIA) performed on the ARCHITECT i System. It measures 1116-NS-19-9 reactive determinants in human serum or plasma. In the first step, patient samples are combined with 1116-NS-19-9 coated paramagnetic microparticles. After washing, an acridinium-labeled 1116-NS-19-9 conjugate is added. Following a second wash, trigger solutions are added to initiate a chemiluminescent reaction, measured as relative light units (RLUs). The RLU output is directly proportional to the concentration of 1116-NS-19-9 reactive determinants. The device is used in clinical and hospital laboratories by trained personnel. Results are interpreted by clinicians alongside other clinical and laboratory data to monitor disease status in patients with confirmed pancreatic cancer. The assay provides quantitative data to assist in managing patient care and assessing disease progression.
Clinical Evidence
Clinical performance was evaluated using 259 serum specimens comparing the ARCHITECT CA 19-9xR assay to the predicate RIA. Passing-Bablok regression showed a correlation coefficient of 0.96. Reproducibility studies (total precision %CV) were ≤ 10%. A serial specimen study of 74 pancreatic cancer patients (261 observations) assessed the association between marker changes and disease state, reporting a total concordance of 61.0% (95% CI: 57.10% - 79.17%) for changes ≥ 14.0%. Reference ranges were established using 360 apparently healthy subjects.
Technological Characteristics
CMIA technology; uses 1116-NS-19-9 mouse monoclonal capture and F(ab')2 conjugate antibodies. Energy source: chemiluminescent reaction measured by ARCHITECT i System optics. Connectivity: integrated with ARCHITECT i System. Software: automated RLU-to-concentration calculation via calibration curve. Form factor: reagent kit for automated analyzer.
Indications for Use
Indicated for the serial measurement of CA 19-9 to aid in the management of patients diagnosed with cancers of the pancreas. Serial testing for patient CA 19-9ₓₗ assay values is used in conjunction with other clinical methods in the management of pancreatic cancer patients.
Regulatory Classification
Identification
A tumor-associated antigen immunological test system is a device that consists of reagents used to qualitatively or quantitatively measure, by immunochemical techniques, tumor-associated antigens in serum, plasma, urine, or other body fluids. This device is intended as an aid in monitoring patients for disease progress or response to therapy or for the detection of recurrent or residual disease.
Special Controls
*Classification.* Class II (special controls). Tumor markers must comply with the following special controls: (1) A guidance document entitled “Guidance Document for the Submission of Tumor Associated Antigen Premarket Notifications (510(k)s) to FDA,” and (2) voluntary assay performance standards issued by the National Committee on Clinical Laboratory Standards.
Predicate Devices
- Fujirebio Diagnostics, Inc. CA 19-9 RIA (K020566)
Related Devices
- K031393 — BAYER ADVIA CENTAUR CA 19-9 ASSAY · Bayer Healthcare, LLC · Jun 24, 2003
- K200997 — Lumipulse G CA19-9-N · Fujirebio Diagnostics,Inc. · May 14, 2020
- K191973 — Lumpipulse G CA19-9-N · Fujirebio Dianostics, Inc. · Oct 22, 2019
Submission Summary (Full Text)
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# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY
A. 510(k) Number:
k052000
B. Purpose for Submission:
New device.
C. Measurand:
CA 19-9
D. Type of Test:
Quantitative, Chemiluminescent Microparticle Immunoassay (CMIA)
E. Applicant:
Fujirebio Diagnostics, Inc.
F. Proprietary and Established Names:
ARCHITECT® CA 19-9™_XR Assay, ARCHITECT® CA 19-9™_XR Calibrator Kit,
ARCHITECT® CA 19-9™_XR Control Kit
G. Regulatory Information:
1. Regulation section:
21 CFR 866.6010, Tumor-associated antigen immunological test system
21 CFR 862.1150, Calibrator
21 CFR § 862.1660, Quality Control Material (Assayed and Unassayed)
2. Classification:
Class II, CA 19-9 assay and Calibrator
Class I, Quality control material
3. Product Code:
NIG, System, Test, Carbohydrate antigen (CA 19-9) for monitoring and
management of pancreatic cancer;
JIT, Calibrator, Secondary
JJX, Single (Specified) Analyte Controls (Assayed and Unassayed)
4. Panel:
Immunology (82)
Chemistry (75), Calibrator and Quality control material
H. Intended Use:
1. Intended use(s):
The ARCHITECT® CA 19-9™_XR assay is a Chemiluminescent Microparticle
Immunoassay (CMIA) for the quantitative determination of 1116-NS-19-9
reactive determinants inhuman serum and plasma on the ARCHITECT i
System. The ARCHITECT® CA 19-9™_XR assay is to be used as an aid in the
management of pancreatic cancer patients with a detectable level of Ca 19-9 at
some point in their disease process and in conjunction with other clinical
methods.
The ARCHITECT® CA 19-9™_XR Calibrators are for the calibration of the
ARCHITECT i System when used for the quantitative determination of 1116-
19-9 reactive determinants in human serum and plasma. Refer to the
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ARCHITECT® CA 19-9™ₓₗ reagent package insert for additional information.
The ARCHITECT® CA 19-9™ₓₗ Controls are for the estimation of test precision and the detection of systematic analytical deviations of the ARCHITECT i System (reagents, calibrators, and instrument), when used for the quantitative determination of 1116-NS-19-9 reactive determinants in human serum and plasma. Refer to the ARCHITECT® CA 19-9™ₓₗ reagent package insert for additional information.
2. Indication(s) for use:
Indicated for the serial measurement of CA 19-9 to aid in the management of patients diagnosed with cancers of the pancreas. Serial testing for patient CA 19-9ₓₗ assay values is used in conjunction with other clinical methods in the management of pancreatic cancer patients.
3. Special condition for use statement(s):
Patients known to be genotypically negative for Lewis blood group antigen are unable to produce the CA 19-9 antigen even in the presence of malignant tissue. Phenotyping for the presence of the Lewis blood group antigen may be insufficient to detect true Lewis antigen negative individuals. Even patients who are genotype positive for the Lewis antigen may produce varying levels of CA 19-9 as the result of gene dosage effect.
4. Special instrument Requirements:
ARCHITECT i Systems – ARCHITECT i 2000 and ARCHITECT i 2000ₓₗ. Both systems belong to the ARCHITECT family of instruments. The ARCHITECT i 2000ₓₗ is similar to the ARCHITECT i 2000 but has the following additional features a) STAT sampling hardware and software, b) Auto Retesting software and c) different composition and position of the RV loader.
Device Description:
The ARCHITECT® CA 19-9™ₓₗ assay consists of:
1. Microparticles coated with monoclonal mouse anti-116-NS-19-9 antibodies in citrate buffer with bovine protein stabilizers and antimicrobial agent.
2. Acridinium-labeled monoclonal mouse anti-1116-NS-19-9 antibody conjugate in phosphate buffer with bovine protein stabilizers and antimicrobial agent.
The following reagents are required but not provided with the ARCHITECT® CA 19-9™ₓₗ assay kit:
The ARCHITECT® CA 19-9™ₓₗ Calibrator Kit consists of:
1. Calibrator A is a TRIS buffer with bovine protein stabilizers and antimicrobial agent
2. Calibrators B to F are preparations of 1116-NS-19-9 reactive determinants (human) in TRIS buffer with bovine protein stabilizers and antimicrobial agent with CA 19-9 antigen concentrations of 30, 100, 250, 600 and 1200 U/mL.
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The ARCHITECT® CA 19-9™ $_{XR}$ Control Kit consists:
1. Control L with target CA 19-9 concentration of 40 U/mL (ranges from 26.0-54.0 U/mL)
2. Control M with target CA 19-9 concentration of 150 U/mL (ranges from 102-198 U/mL)
3. Control H with target CA 19-9 concentration of 750 U/mL (ranges from 510 - 990 U/mL).
These controls are preparations of human CA 19-9 defined antigen in TRIS buffer with bovine protein stabilizers and antimicrobial agent.
ARCHITECT i Pre-Trigger Solution
ARCHITECT i Trigger Solution
ARCHITECT i Wash Buffer
ARCHITECT i Multi-Assay Manual Diluent.
## J. Substantial Equivalence Information:
1. Predicate device name(s): Fujirebio Diagnostics CA 19-9™ RIA
2. Predicate K number(s): k020566
3. Comparison with predicate:
| Similarities | | |
| --- | --- | --- |
| Item | Device | Predicate |
| | ARCHITECT® CA 19-9™_{XR} | Fujirebio CA 19-9 RIA |
| Intended Use | Quantitative analysis of CA 19-9 in human serum and plasma | Same |
| Indications for Use | As an aid in management of patients with cancers of the exocrine pancreas | Same |
| Antibody Type and Source | Monoclonal, mouse | Same |
| Differences | | |
| --- | --- | --- |
| Item | Device | Predicate |
| Methodology | Chemiluminescent Microparticle Immunoassay | Radioimmunoassay |
| Sample type | Serum and plasma (EDTA, lithium heparin and sodium heparin) | Serum or plasma (Citrate, heparin and EDTA) |
| Capture | Monoclonal anti-CA 19-9 antibody coated paramagnetic microparticles | Mouse monoclonal anti-CA 19-9 coated polystyrene beads |
| Conjugate Antibody | Acridinium labeled monoclonal anti-CA 19-9 F(ab')2 conjugate | 125I conjugated monoclonal anti-CA 19-9 antibody |
| Calibrators | 6 levels (0 - 1200 U/mL) | 6 levels (0-240 U/mL) |
| Controls | 3 levels every 24 hours
Low = 40 U/mL | 2 levels each run
Low = 40 - 50 U/mL |
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| Differences | | |
| --- | --- | --- |
| Item | Device | Predicate |
| | Medium = 150 U/mL High = 750 U/mL) | High = 80 - 90 U/mL |
| Instrument System | ARCHITECT i System | Manual method |
| Measuring range | 0-1200 U/mL | 0-240 U/mL |
| Reaction time | ≤ 30 minutes | 6 hours |
| Automated dilution | Yes | No |
| Calibration frequency | Every 30 days | Every run |
| Sample size | < 100 μL | 200 μL |
# K. Standard/Guidance Document Referenced (if applicable):
Special guidance "Guidance Document for the Submission of Tumor Associated Antigen premarket Notifications (510(k)s) to FDA". CLSI guidelines include EP5-A (Evaluation of Precision Performance of Clinical Chemistry Devices), EP7-A (Interference Testing in Clinical Chemistry Devices), EP9-A2 (Method Comparison and Bias Estimation Using Patient Samples), EP6-P2 (Evaluation of the Linearity of Quantitative Analytical Methods - Proposed Guideline), C28-A2 (How to Define and Determine reference Intervals in the Clinical Laboratory) and EP14-A (Evaluation of Matrix Effects).
# L. Test Principle:
The ARCHITECT® CA 19-9™ $_{XR}$ assay is a two-step immunoassay to determine the presence of 1116-NS-19-9 reactive determinants in human serum or plasma, using CMIA technology with flexible assay protocols, referred to as Chemiflex®. In the first step of the assay, sample and anti-CA 19-9 coated paramagnetic microparticles are combined. CA 19-9 reactive determinants present in the sample bind to the anti-CA 19-9 coated microparticles. After washing, anti-CA 19-9 acridinium-labeled conjugate is added in the second step. After another wash cycle, pre-trigger and trigger solutions are then added to the reaction mixture; the resulting chemiluminescent reaction is measured as relative light units (RLUs). A direct relationship exists between the quantity of CA 19-9 reactive determinants in the sample and the RLUs detected by the ARCHITECT $i$ optical system.
# M. Performance Characteristics (if/when applicable):
# 1. Analytical performance:
# i. Precision/Reproducibility:
Precision was evaluated according to CLSI EP5-A2. Two studies were performed:
Study 1 - Six samples (2 serum pools [Panel 1 and 2], 1 CA 19-9 spiked serum pool [Panel 3] and ACHITECT® CA $19 - 9^{\mathrm{TM}}_{XR}$ Controls) were tested in duplicate, two runs per day for 20 nonconsecutive days on two instruments and two lots of ACHITECT® CA $19 - 9^{\mathrm{TM}}_{XR}$ reagents. Results for within-run and total precision are summarized below.
| Sample | Reagent Lot | Instrument | N | Mean Conc. (U/mL) | Within-run | | Total | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | | | | SD (U/mL) | %CV | SD (U/mL) | %CV |
| Panel 1 | 1 | 1 | 80 | 56.52 | 1.69 | 3.0 | 2.19 | 3.9 |
| | 2 | 2 | 80 | 51.20 | 1.80 | 3.5 | 2.10 | 4.1 |
| Panel 2 | 1 | 1 | 80 | 311.49 | 7.22 | 2.3 | 10.72 | 3.4 |
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| Sample | Reagent Lot | Instrument | N | Mean Conc. (U/mL) | Within-run | | Total | |
| --- | --- | --- | --- | --- | --- | --- | --- | --- |
| | | | | | SD (U/mL) | %CV | SD (U/mL) | %CV |
| Panel 3 | 2 | 2 | 80 | 288.82 | 9.14 | 3.2 | 11.23 | 3.9 |
| | 1 | 1 | 80 | 744.81 | 27.82 | 3.7 | 36.85 | 5.0 |
| Low Control | 2 | 2 | 80 | 728.82 | 42.53 | 5.8 | 47.66 | 6.5 |
| | 1 | 1 | 80 | 45.03 | 2.59 | 5.8 | 2.98 | 6.6 |
| Med Control | 2 | 2 | 80 | 42.33 | 2.94 | 6.9 | 3.60 | 8.5 |
| | 1 | 1 | 80 | 157.66 | 5.99 | 3.8 | 8.52 | 5.4 |
| High Control | 2 | 2 | 80 | 146.93 | 6.26 | 4.3 | 8.14 | 5.5 |
| | 1 | 1 | 80 | 781.68 | 44.76 | 5.7 | 49.87 | 6.4 |
| | 2 | 2 | 80 | 781.42 | 62.10 | 8.0 | 65.28 | 8.4 |
Study 2 – Four representative human serum pools spiked with various concentrations of CA 19-9 (50.06, 288.16, 719.38 and 677.26 U/mL) were tested in replicates of four, two runs per day for 13 nonconsecutive days on two instruments with two lots of reagents. The total %CV ranged from 3.7% to 6.2%.
ii. Linearity/assay reportable range:
Linearity was evaluated according to CLIS EP6-P2. Aliquots of 10 human serum samples were spiked with CA 19-9 to concentrations within the assay dynamic range. Each sample was automatically diluted by the ARCHITECT instrument with wash buffer. The undiluted and diluted samples were assayed in duplicate and the percent recoveries determined. The expected value was obtained by dividing the undiluted observed value by a dilution factor. The average percent recovery of each sample ranged from 98% to 117% with a mean average of 105%. Acceptance criterion for mean recovery was 100±15%.
Auto-dilution verification was assessed by assaying aliquots of 10 human serum samples spiked with CA 19-9 to concentrations within the assay dynamic range and manually diluted 1:10 with the wash buffer. The undiluted and diluted samples were tested in replicates of two. The undiluted samples were also tested in replicates of two using the 1:10 auto-dilution protocol of the assay. The percent recoveries of the manual and auto-diluted samples were calculated and compared. The average recovery of the auto-dilution protocol to manual dilution was 93%.
Spike recovery – Aliquots of 10 normal human serum specimens with known endogenous CA 19-9 levels were spiked with various concentrations of CA 19-9 (124.2, 175.7, 629.9 and 884.7 U/mL) and assayed in duplicate. The percent recovery was calculated. The percent recoveries ranged from 94% to 128% and the mean recovery was 105%.
The assay measuring range is from 0 U/mL to 1200 U/mL.
iii. Traceability, Stability, Expected values (controls, calibrators, or methods):
There is no known reference standard for CA 19-9. The ARCHITECT® CA 19-9™XR Calibrators were standardized against the Fujirebio Diagnostics, Inc. CA 19-9 reference preparation. The ARCHITECT® CA 19-9™XR primary calibrators are manufactured gravimetrically by diluting the CA 19-9 antigen stock in Calibrator Standard Matrix and assayed
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using the Fujirebio Diagnostics Inc. CA 19-9™ RIA assay. The values were validated by a panel of patient serum samples on two lots of ARCHITECT® CA 19-9™ $_{XR}$ reagent kit and three ARCHITECT instrument systems. The same panel was also tested on three lots of the Fujirebio Diagnostics Inc. CA 19-9™ RIA kit.
# iv. Detection limit (functional sensitivity):
The minimal detectable dose (MDD) was determined by testing the ARCHITECT® CA 19-9™ $_{XR}$ Calibrator A (0 U/mL) in replicates of 10 followed by two replicates of ARCHITECT® CA 19-9™ $_{XR}$ Calibrator B (30 U/mL), on three instruments using two lots of calibrators and two lots of ARCHITECT® CA 19-9™ $_{XR}$ reagents. The mean values and the standard deviations (SD) of the 18 sets of Calibrator A and the mean values of the 18 sets of Calibrator B were used to calculate the MDD for each run. MDD = [(2 x SD $_{\text{CalA}}$ ) x Conc $_{\text{CalB}}$ ]/(Mean RLU $_{\text{CalB}}$ - Mean RLU $_{\text{CalA}}$ ). Limit of detection (LOD) is defined as the lowest measurable CA 19-9 concentration that can be distinguish from zero and equal to (MDD + 2SD). The mean LDL claim is ≤ 2.0 U/mL.
| Number of runs (n) | Mean MDD (U/mL) | MDD SD | MDD Range (U/mL) | LOD (Mean MDD + 2SD) |
| --- | --- | --- | --- | --- |
| 18 | 0.48 | 0.26 | 0.25 – 1.28 | 1.00 |
# v. Analytical specificity:
Endogenous substances - Interference was determined by spiking a known amount of an interfering substance into serum samples supplemented with CA 19-9. Interfering substances tested included hemoglobin (600 mg/dL), bilirubin (22 mg/dL), triglycerides (5100 mg/dL) and total protein (10 g/dL). Five samples were used for bilirubin, hemoglobin and triglycerides and four samples for total protein. Spiked and non-spiked samples were tested in duplicate and percent recoveries were calculated. Percent mean recoveries ranged from 91% for total protein to 102% for hemoglobin at the levels tested.
Pharmaceutical compounds - Interference was determined by spiking the following pharmaceutical compounds into serum samples and assayed: Leucovorin (11.4 mg/dL), Gemzar/Gemcitabine HCl (38.2 mg/dL), Streptozotocin/Zanosar (28 mg/dL), Doxorubicin (4 mg/dL). Cyclophosphamide/Cytoxan (37.5 mg/dL), Cisplatin (5.7 mg/dL), 5-Flourouracil/Adrucil (39 mg/dL), Methotrexate/Amethopterin-Hydrate (91 mg/dL), Tamoxifen (2.28 μg/dL), Cytarabine (3 mg/dL) and Paclitaxel/Taxol (6.7 mg/dL). Spiked and non-spiked samples were tested in replicates and percent recoveries were calculated. At the concentrations tested, the percent mean recoveries ranged from 97% to 104%.
Human anti-mouse antibody (HAMA) - To assess interference due to HAMA, five HAMA positive samples and one normal sample were studied. Each sample was split into three aliquots. One aliquot was
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spiked with CA 19-9 antigen to achieve 35 U/mL while the second aliquot was spiked with the same volume of antigen to achieve 250 U/mL. The third aliquot was spiked with an equivalent volume of antigen free matrix and served as the control. All aliquots were run in duplicate in the same run. Percent recoveries were calculated and for the HAMA samples with 35 U/mL CA 19-9, the % recovery ranged from 84% to 95% (mean = 91%) and for samples with 250 U/mL CA 19-9, the % recovery ranged from 74% to 104% (mean = 94%).
**Rheumatoid factor (RF)** - To assess interference due to RF, five RF positive samples and one normal sample were tested. Each sample was split into three aliquots. One aliquot was spiked with CA 19-9 antigen to achieve 35 U/mL while the second aliquot was spiked with the same volume of antigen to achieve 250 U/mL. The third aliquot was spiked with an equivalent volume of antigen free matrix and served the control. All aliquots were run in duplicate in the same run. Percent recoveries were calculated and recoveries for samples with 35 U/mL CA 19-9 ranged from 82% to 95% (mean = 88%) and for samples with 250 U/mL CA 19-9, % recovery ranged from 74% to 104% (mean = 99%).
**Cross-reactivity**
No data provided.
vi. Assay cut-off:
See Expected Value.
2. Comparison studies:
i. Method comparison with predicate device:
Two hundred fifty nine serum samples were tested on the ARCHITECT® CA 19-9™XR assay and the Fujirebio CA 19-9™ RIA Assay. All samples with values outside the dynamic range of either assay were diluted per package inserts. Excluded from analysis were 41 samples with values below the limit of detection (< 2.0 U/mL) and 23 samples with values above the dynamic range even after auto dilution (> 12,000 U/mL) of the ARCHITECT® CA 19-9™XR assay. In addition, one sample with discrepant results on both assays and one with an invalid CV in the Fujirebio assay were excluded. The remaining 193 samples were analyzed by Passing-Bablok linear regression analysis. The CA 19-9 concentrations of the specimen as determined by the Fujirebio CA 19-9™ RIA ranged from 1.17 to 10,782 U/mL. The Passing-Bablok linear regression analysis yielded a slope of 1.2 (99% CI 1.08, 1.37) and y-axis intercept of -5.1 U/mL (99% CI -7.4, -3.4) with a Spearman Correlation coefficient of 0.96.
In addition to regression analysis, samples were also evaluated to the percent agreement with the predicate device (see table below):
| | CA 19-9 RIA | | | |
| --- | --- | --- | --- | --- |
| | | ≤ 37 U/mL | > 37 U/mL | Total |
| Architect CA 19-9 | ≤ 37 U/mL | 151 | 4 | 155 |
| | > 37 U/mL | 3 | 76 | 79 |
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# Comparison of ARCHITECT CA 19-9 $_{XR}$ i2000 and i2000 $_{SR}$
Analytical sensitivity - Ten replicates of calibrator A and two replicates of calibrator B were tested per run for six runs on one $i2000_{SR}$ instrument using one calibrator lot and two reagent lots. Analytical sensitivity was $0.60\mathrm{U / mL}$ . The same samples were also assayed on two $i2000$ instruments for 12 runs. The analytical sensitivity for the $i2000$ was $1.14\mathrm{U / mL}$ . Analytical sensitivity for both instrument met the acceptance criterion of $\leq 2.0\mathrm{U / mL}$ .
Precision - Three serum panels and three assay controls (low, medium and high) were tested in duplicate per run, two runs per day for 20 nonconsecutive days on one $i2000$ and one $i2000_{SR}$ instrument. Results are summarized below and both systems met the acceptance criterion of total CV of $\leq 10\%$ .
| Sample | Total CV (%) | |
| --- | --- | --- |
| | i2000 | i2000SR |
| Control L | 6.6 | 8.5 |
| Control M | 5.4 | 5.5 |
| Control H | 6.4 | 8.4 |
| Panel 1 | 3.9 | 4.1 |
| Panel 2 | 3.4 | 3.9 |
| Panel 3 | 4.9 | 6.5 |
Spike recovery - Ten normal human serum samples each spiked with four different concentrations of CA 19-9 (125, 175, 650 and $900\mathrm{U / mL}$ ) were tested. Five of the samples were tested on one $i2000$ instrument and the other five samples on one $i2000_{SR}$ instrument. Each sample was assayed in duplicate. The mean percent recovery across all concentrations and all samples on the $i2000$ instrument was $98\%$ as compared to $111\%$ on the $i2000_{SR}$ instrument. Both instruments met the acceptance criterion of $100 \pm 15\%$ .
Correlation - One hundred twenty samples were assayed on both instruments. The acceptance criteria were a slope of $1.0 \pm 0.2$ for samples with CA 19-9 values between $0 - 100\mathrm{U / mL}$ and a r-value of $\geq 0.90$ for samples with CA 19-9 values of $0 - 1200\mathrm{U / mL}$ . Regression results are summarized in table below.
| CA 19-9 (U/mL) | N | R | Least Squares | | Passing-Bablok | |
| --- | --- | --- | --- | --- | --- | --- |
| | | | Slope | Intercept | Slope | Intercept |
| 0-100 | 92 | 0.98 | 0.95 | 1.36 | 0.99 | 0.54 |
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ii. Matrix comparison:
Matched human serum and plasma samples were collected in the following tube types: serum clot, serum separator tube (SST), EDTA, lithium heparin and sodium heparin. Twenty-six sample sets were assayed unchanged. Twenty-four sample sets were subdivided into 4 groups and each group was spiked with CA 19-9 to achieve concentrations of 50, 100, 200 or 650 U/mL. All samples were tested in duplicate and within 36 hours of sample draw. Results of the average recovery for each anticoagulant are summarized below.
| CA 19-9 (U/mL) | Average % Recovery | | | |
| --- | --- | --- | --- | --- |
| | SST | EDTA | Sodium Heparin | Lithium Heparin |
| 50 | 102 | 96 | 103 | 102 |
| 100 | 99 | 101 | 101 | 102 |
| 200 | 101 | 98 | 99 | 95 |
| 650 | 103 | 101 | 101 | 102 |
| Total % and (SD)* | 100 (5) | 98 (5) | 100 (6) | 100 (7) |
| CA 19-9 (U/mL) | Average U/mL Recovery and (SD) | | | |
| <24 | 0.7 (1.4) | 0 (0.8) | 0 (0.9) | 0 (0.9) |
*Includes samples between 24 to 50 U/mL
3. Clinical studies:
a. Clinical sensitivity: Not applicable.
b. Clinical specificity: Not applicable.
c. Other clinical supportive data (when a. and b. are not applicable): Serial Monitoring Analysis
Two hundred and sixty one serum samples from 74 patients with confirmed pancreatic cancer collected and banked at two US clinical sites were analyzed. The average number of sample pairs per patient was 3.5 (see table below)
| Number in Series | Number of Observation Pairs | Frequency | Percent |
| --- | --- | --- | --- |
| 3 | 2 | 36 | 48.6 |
| 4 | 3 | 37 | 50.0 |
| 5 | 4 | 1 | 1.4 |
The average age of the patients at the time of diagnosis was 61.8 years ranging from 41 to 85 years. Fifty-five percent were males and forty-five percent females. Majority of the patients (80%) were Caucasians, the remaining 20% consisted of African-Americans (11%), Hispanics (8%) and Asians (1%). Only 9% of the patients were current smokers, 51% were past smokers and 38% nonsmokers (smoking status of one patient was unknown). At the time of diagnosis, 30% of the patients had diabetes and 27% had other medical conditions. Of the diabetic patient subset, 64% were males.
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Sixty-nine of the 74 patients had disease stage information: 4.34% were Stage I, 7.25% Stage II, 44.9% Stage III, and 43.5% stage IV. Ninety-five percent of the pancreatic tumors were adenocarcinomas with 65% involving the head of the pancreas. In addition, histology stratification showed that 13.51% were as well differentiated, 22.97% moderately differentiated, 5.41% poor to moderately differentiated, 12.16% poorly differentiated and 45.95% as other.
Changes in CA 19-9 concentrations and changes in disease state were analyzed on a per-visit basis. A significant change in CA 19-9 was defined as greater than 14% (2.5 times the total precision %CV). The following tables show the association between CA 19-9 concentrations and disease status for the 187 evaluable observation pairs. The 95% confidence intervals for the concordance statistics were based on General Estimable Equations and calculated using the GENMOD procedure of SAS.
| | Change in Disease State | | |
| --- | --- | --- | --- |
| Changes in CA 19-9 | Progression | No Progression | Total |
| ≥14% | 16 | 56 | 72 |
| < 14% | 17 | 98 | 115 |
| Total | 33 | 154 | 187 |
Positive concordance = 0.485 (16/33) (95% CI: 0.318, 0.655)
Negative concordance = 0.636 (98/154) (95% CI: 0.560, 0.706)
Total concordance = 0.61 (114/187) (95% CI: 0.543, 0.673)
| Fujirebio RIA | Change in Disease State | | |
| --- | --- | --- | --- |
| Changes in CA 19-9 | Progression | No Progression | Total |
| ≥ 20% | 15 | 51 | 66 |
| < 20% | 18 | 103 | 121 |
| Total | 33 | 154 | 187 |
Positive concordance = 0.455 (15/33) (95% CI: 0.284, 0.637)
Negative concordance = 0.669 (103/154) (95% CI: 0.585, 0.743)
Total concordance = 0.631 (118/187) (95% CI: 0.554, 0.702)
Serial monitoring results were also analyzed on a per-patient basis as shown below. Concordances and 95% CI were determined. Confidence intervals for these estimates were determined using binomial distribution.
| | Change in Disease State | | |
| --- | --- | --- | --- |
| Changes in CA 19-9 | Progression | No Progression | Total |
| ≥ 14% | 15 | 16 | 31 |
| < 14% | 7 | 36 | 43 |
| Total | 22 | 52 | 74 |
Positive concordance = 0.682 (15/22) (95% CI: 0.451, 0.861)
Negative concordance = 0.692 (36/52) (95% CI: 0.549, 0.813)
Total concordance = 0.689 (51/74) (95% CI: 0.571, 0.792)
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| Fujirebio RIA | Change in Disease State | | |
| --- | --- | --- | --- |
| Changes in CA 19-9 | Progression | No Progression | Total |
| ≥ 20% | 14 | 18 | 32 |
| < 20% | 8 | 34 | 42 |
| Total | 22 | 52 | 74 |
Positive concordance = 0.636 (14/22) (95% CI: 0.407, 0.828)
Negative concordance = 0.654 (34/52) (95% CI: 0.509, 0.780)
Total concordance = 0.649 (48/74) (95% CI: 0.529, 0.756)
4. Clinical cut-off:
Not applicable.
5. Expected values/Reference range:
The normal reference range was established by testing serum samples from 360 apparently healthy subjects consisted of 180 females and 180 males. Of the 180 female subjects, 56 were post-menopausal and 124 premenopausal. No age or ethnicity information was provided. No analysis was performed according to age, gender or ethnicity. A cumulative distribution was established and the $99^{\text{th}}$ percentile was determined to be $64.39\mathrm{U/mL}$ and the $95^{\text{th}}$ percentile was $37.83\mathrm{U/mL}$ . The table below shows the distribution of CA 19-9 results.
| # subjects | Percent (%) | | | | |
| --- | --- | --- | --- | --- | --- |
| | 0-37.0 U/mL | 37.1-100 U/mL | 100.1-500 U/mL | 500.1-1200 U/mL | >1200 U/mL |
| 360 | 94.4 | 5.6 | 0.0 | 0.0 | 0.0 |
In addition to the normal cohort, 441 serum samples from patients with benign conditions and 537 from patients with malignant diseases were tested. The following table summarizes the sample distribution, diseases/conditions and distribution of CA 19-9 results.
| Cohort | # Subjects | Distribution of CA 19-9 Values (%) | | | | |
| --- | --- | --- | --- | --- | --- | --- |
| Nonmalignant Disease | | 0-37.0 U/mL | 37.1-100 U/mL | 100.1-500 U/mL | 500.1-1200 U/mL | >1200 U/mL |
| Rectal Polys | 33 | 97 | 3 | 0 | 0 | 0 |
| Pancreatitis | 3 | 100 | 0 | 0 | 0 | 0 |
| Gall bladder | 21 | 95.2 | 0 | 0 | 0 | 4.8 |
| Diabetes | 38 | 94.7 | 5.3 | 0 | 0 | 0 |
| Pulmonary | 40 | 100 | 0 | 0 | 0 | 0 |
| Cirrhosis | 153 | 92.8 | 4.6 | 0.7 | 0.7 | 1.3 |
| Hepatitis | 68 | 92.6 | 7.4 | 0 | 0 | 0 |
| Renal | 34 | 91.2 | 8.8 | 0 | 0 | 0 |
| Other | 51 | 96.1 | 3.9 | 0 | 0 | 0 |
| Malignant Disease | | | | | | |
| Colorectal | 169 | 81.1 | 7.7 | 5.3 | 1.2 | 4.7 |
| Pancreatic | 66 | 43.9 | 6.1 | 12.1 | 10.6 | 27.3 |
| Gastric | 69 | 66.7 | 11.6 | 10.1 | 2.9 | 8.7 |
| Hepatocellular | 30 | 63.3 | 16.7 | 3.3 | 10.0 | 6.7 |
| Pulmonary | 70 | 84.3 | 5.7 | 4.3 | 1.4 | 4.3 |
| Mammary | 102 | 86.3 | 10.8 | 2.0 | 1.0 | 0 |
| Ovarian | 31 | 87.1 | 6.5 | 3.2 | 3.2 | 0 |
N. Proposed Labeling:
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The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10.
**O. Conclusion:**
The submitted information in this premarket notification is complete and supports a substantial equivalence decision.
