(90 days)
For in vitro diagnostic use with the IMMULITE® 2000 Systems Analyzers - for the quantitative measurement of CA15-3 antigen in human serum and plasma, as an aid in the detection of recurrence in previously treated stage III breast cancer patients, and in the management of metastatic breast cancer patients by monitoring disease progression or response to treatment. Serial testing for patient CA15-3 values should be used in conjunction with other clinical methods used for detecting early recurrence in stage III disease and for monitoring response to treatment in patients with metastatic breast cancer.
The IMMULITE® 2000 BR-MA assay was cleared under K013984. The components of the cleared assay were modified to reduce biotin interference. The modified IMMULITE® 2000 BR-MA Assay is comprised of the following components: BR-MA Bead Pack (L2BR12), BR-MA Reagent Wedge (L2BRA2) - Well 1, BR-MA Reagent Wedge (L2BRA2) - Well 2, and BR-MA Adjustors (LBRL, LBRH). The IMMULITE 2000 BR-MA is a solid-phase, two-step chemiluminescent immunometric assay. There are two incubation cycles of 30 minutes each. During the initial 30-minute cycle, the patient sample is incubated with biotinylated antibody coated bead (bead pack) and a buffer (reagent wedge well 1). The biotinylated antibody on the bead captures the antigen in the patient sample. On completion of the first 30-minute cycle, unbound sample/buffer are then removed via a centrifugal wash. During the second 30-minute cycle, alkaline phosphatase antibody conjugate in buffer (reagent wedge well 2) is added to complete the bead pair immunocomplex sandwich consisting of capture Ab-antigen-detection Ab. On completion of the second 30-minute cycle, unbound conjuqate is removed by centrifugal wash. The amount of alkaline phosphatase bound is directly proportional to the patient sample. Following the two 30-minute incubation periods. IMMULITE chemiluminescent substrate (L2SUBM) is added for a further 5-minute incubation period to generate the luminogenic reaction. The chemiluminescent substrate undergoes hydrolysis in the alkaline phosphatase to yield an unstable intermediate, which then emits photons. The sustained emissions are measured by the luminometer. The resulting relative light units are proportional to the concentration of CA15-3 in the sample, which is expressed as U/mL.
The retrieved document describes the acceptance criteria and performance of the IMMULITE 2000 BR-MA assay, which is a tumor-associated antigen immunological test system for CA15-3. The modifications to the device primarily focus on reducing biotin interference.
Here's a breakdown of the requested information based on the provided text:
1. Table of Acceptance Criteria and Reported Device Performance
The document doesn't explicitly list "acceptance criteria" against "reported device performance" in a single table. Instead, it presents performance characteristic studies that implicitly demonstrate the device meets certain standards. I've aggregated these into a table format below, using typical performance metrics for such devices. The "Acceptance Criteria" are implied by common laboratory standards (e.g., CLSI guidelines) and the successful outcome of the tests.
| Performance Metric | Implied Acceptance Criteria | Reported Device Performance |
|---|---|---|
| Detection Limits | Determined in accordance with CLSI EP17-A2. | LoB = 0.21 U/mL, LoD = 0.30 U/mL, LoQ = 1 U/mL. |
| Linearity/Measuring Interval | Linearity across the assay range, ADL ≤ 15% at each level. | Confirmed across the assay range (1 - 300 U/mL) with acceptable ADL at each individual level. |
| Method Comparison | Strong correlation with the currently marketed device. | N=274 serum samples. Correlation Coefficients: Lot 1 ($Y = 0.98x + 0.71$) = 0.989; Lot 2 ($Y = 0.99x + 0.16$) = 0.992; Lot 3 ($Y = 1.04x - 0.79$) = 0.990. Statistical method: Passing-Bablok regression. |
| Assay Precision (Within-Lab) | %CV within acceptable limits for the assay. | 5 serum samples tested. %CV ranged from 7.1% to 7.4% (Total/Within-Lab). Within-Run %CV ranged from 4.8% to 6.8%. |
| Assay Reproducibility | %CV within acceptable limits across multiple lots and days. | 5 serum samples tested across 3 reagent lots. Total Reproducibility %CV ranged from 4.5% to 6.0%. |
| Recovery | Expected recovery within an acceptable range (e.g., 90-110%). | % Recovery for spiked samples ranged from 92% to 105%. |
| Interference | No significant interference from tested endogenous/exogenous substances up to specified concentrations. | No significant interference observed for Hemoglobin (381 mg/dL), Conjugated and Unconjugated Bilirubin (200 mg/L), Intralipid (3000 mg/dL), Biotin (3500 ng/mL), and several chemotherapy drugs (e.g., 5-Fluorouracil 1000 µg/mL). Note: The key improvement is reduced biotin interference from 100 ng/mL (predicate) to 3500 ng/mL. |
| Cross-Reactivity | No detectable cross-reactivity with specified tumor markers. | No detectable specificity (cross-reactivity) for Alpha-fetoprotein, CA125, CA19-9, and Carcinoembryonic Antigen at high concentrations. |
| Hook Effect | No hook effect within the assay range and beyond. | No hook effect observed up to 80,000 U/mL (well above the measuring interval of 300 U/mL). |
| Reference Range Verification | Verification of existing reference range with healthy samples. | 94% (65 out of 69) of normal female samples fell within the existing reference range (6.4 - 58 U/mL) across three lots. |
| Matrix Comparison | Comparable values across different specimen types. | Comparable values demonstrated for serum, Lithium Heparin, and EDTA plasma samples. |
2. Sample Size Used for the Test Set and Data Provenance
- Detection Limits (LoB, LoD, LoQ): The specific sample size for determining LoB, LoD, and LoQ is not explicitly stated as a number of individual patient samples, but the study was conducted "in accordance with CLSI EP17-A2," which provides methodologies for these determinations.
- Linearity/Measuring Interval: A high and low sample pool were used to prepare a panel of ten levels. The number of individual patient samples contributing to these pools is not specified.
- Method Comparison: 274 patient samples.
- Assay Precision: Five serum samples. Each tested in duplicate over 20 days, two runs per day (total 80 replicates per sample). Total N for data points is 400 (5 samples * 80 replicates).
- Assay Reproducibility: Five serum samples. Each tested over 5 days, with 5 replicates per sample (total 25 replicates per sample) across 3 reagent lots. Total N of data points is 375 (5 samples * 25 replicates * 3 reagent lots).
- Recovery: 5 neat samples spiked with 3 different concentrations of CA15-3 solution.
- Interference: Not specified as a number of patient samples, but various compounds were tested.
- Hook Effect: Not specified as a number of patient samples.
- Reference Range Verification: 69 apparently healthy female samples across 3 lots (total 207 results).
- Matrix Comparison: Not explicitly specified, but "comparable values to serum samples" were demonstrated across SST, Lithium Heparin, and EDTA tube types.
Data Provenance: The document does not specify the country of origin of the data or whether the studies were retrospective or prospective. It refers to human serum and plasma samples and "patient samples" and "apparently healthy female samples" without further geographical or temporal details.
3. Number of Experts Used to Establish the Ground Truth for the Test Set and Their Qualifications
This information is not applicable to this type of device. The IMMULITE 2000 BR-MA is an in vitro diagnostic assay that quantitatively measures CA15-3 antigen. Its performance characteristics are established through analytical studies (e.g., precision, linearity, interference) and comparisons to a predicate device or established laboratory methods, rather than through expert interpretation of outputs to establish a "ground truth" (as might be seen in imaging AI, for example). The ground truth for this device is the actual concentration of the analyte, verified through reference methods or spiked samples with known concentrations.
4. Adjudication Method for the Test Set
Not applicable. As described above, this device's performance is not evaluated through expert adjudication of results but rather by comparing its quantitative measurements to known values or to a predicate device, and assessing its analytical characteristics.
5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study Was Done, and Effect Size of How Much Human Readers Improve with AI vs. Without AI Assistance
Not applicable. This is an in vitro diagnostic assay, not an AI-assisted diagnostic imaging or interpretation system requiring human "readers." The "human reader" concept is not relevant here.
6. If a Standalone (i.e., algorithm only without human-in-the-loop performance) Was Done
The device is an automated in vitro diagnostic assay. Its primary function is to perform quantitative measurements of CA15-3. The performance studies described (detection limits, linearity, precision, etc.) are all standalone performance evaluations of the assay itself, without a "human-in-the-loop" component in the sense of interpreting an output that then needs human review for diagnosis. The device provides a quantitative result (U/mL), which is then used by a clinician in conjunction with other clinical methods. So, yes, the performance data presented are for the standalone analytical performance of the device.
7. The Type of Ground Truth Used
The ground truth used in the performance studies includes:
- Known concentrations: For linearity, recovery, interference, and hook effect studies, samples are often prepared with known concentrations of the analyte or interferents.
- Reference methods/Predicate device: For method comparison, results from the candidate device are compared against results from the legally marketed predicate device.
- Statistical methods: Established CLSI (Clinical and Laboratory Standards Institute) guidelines provide the statistical framework and generally accepted methodologies for determining metrics like LoB, LoD, LoQ, precision, and linearity.
- Clinically defined healthy populations: For reference range verification, samples from "apparently healthy female samples" are used.
8. The Sample Size for the Training Set
The document describes performance studies for a modified in vitro diagnostic assay. These studies are typically for verification and validation, not for "training" an algorithm in the sense of machine learning. There is no mention of a "training set" in the context of algorithm development or machine learning. The studies primarily evaluate the analytical performance of the modified assay components.
9. How the Ground Truth for the Training Set Was Established
As there is no mention of a "training set" for an algorithm, this question is not applicable. The device's mechanism is based on immunometric assay principles using chemical reactions, not on training data for a machine learning model.
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Image /page/0/Picture/0 description: The image shows the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is the FDA logo, with the letters "FDA" in a blue square. To the right of the blue square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.
March 13, 2024
Siemens Healthcare Diagnostics Inc Karlyn Kellogg Regulatory Affairs Professional Glyn Rhonwy, Llanberis Caernarfon Llanberis, Gwynedd LL55 4EL United Kingdom
Re: K233946
Trade/Device Name: IMMULITE 2000 BR-MA Regulation Number: 21 CFR 866.6010 Regulation Name: Tumor-Associated Antigen Immunological Test System Regulatory Class: Class II Product Code: MOI Dated: December 14, 2023 Received: December 14, 2023
Dear Karlyn Kellogg:
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 (the 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 available 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.
Additional information about changes that may require a new premarket notification are provided in the FDA guidance documents entitled "Deciding When to Submit a 510(k) for a Change to an Existing Device"
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K233946 - Karlyn Kellogg
(https://www.fda.gov/media/99812/download) and "Deciding When to Submit a 510(k) for a Software Change to an Existing Device" (https://www.fda.gov/media/99785/download).
Your device is also subject to, among other requirements, the Quality System (QS) regulation (21 CFR Part 820), which includes, but is not limited to, 21 CFR 820.30, Design controls; 21 CFR 820.90, Nonconforming product; and 21 CFR 820.100. Corrective and preventive action. Please note that regardless of whether a change requires premarket review. the OS regulation requires device manufacturers to review and approve changes to device design and production (21 CFR 820.30 and 21 CFR 820.70) and document changes and approvals in the device master record (21 CFR 820.181).
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 Part 803) for devices or postmarketing safety reporting (21 CFR Part 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safetyreporting-combination-products); good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR Part 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR Parts 1000-1050.
Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 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 medical devices and radiation-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 (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).
Sincerely,
Ying Mao -S
Ying Mao, Ph.D. Branch Chief Division of Immunology and Hematology Devices OHT7: Office of In Vitro Diagnostics Office of Product Evaluation and Quality Center for Devices and Radiological Health
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Indications for Use
510(k) Number (if known) K233946
Device Name IMMULITE 2000 BR-MA
Indications for Use (Describe)
For in vitro diagnostic use with the IMMULITE 2000 Systems Analyzers - for the quantitative measurement of CA15-3 antigen in human serum and plasma, as an aid in the detection of recurrence in previously treated stage III breast cancer patients, and in the management of metastatic breast cancer patients by monitoring disease progression or response to treatment. Serial testing for patient CA15-3 values should be used in conjunction with other clinical methods used for detecting early recurrence in stage III disease and for monitoring response to treatment in patients with metastatic breast cancer.
| Type of Use (Select one or both, as applicable) | |
|---|---|
| Prescription Use (Part 21 CFR 801 Subpart D) | Over-The-Counter Use (21 CFR 801 Subpart C) |
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510(k) Summary
This 510(k) summary of safety and effectiveness information is submitted in accordance with the requirements of 21 CFR 807.92 and SMDA 1990.
The assigned 510(k) number is: _______________________________________________________________________________________________________________________________________________ K233946
I. Submitter
| Contact Person: | Karlyn Kellogg |
|---|---|
| Address: | Siemens Healthcare Diagnostics Inc.500 GBC Drive, M/S 514Newark, DE 19714 |
| E-mail: | karlyn.e.kellogg@siemens-healthineers.com |
| Phone: | 302-729-6271 |
| Date of Preparation: | March 6, 2024 |
II. Device
| Trade Name: | IMMULITE® 2000 BR-MA |
|---|---|
| Common Name: | System, Test, Immunological, Antigen, Tumor |
| Classification Name: | Tumor-associated antigen immunological test system |
| Regulation Number: | 21 CFR 866.6010 |
| Classification: | Class II |
| Product Code: | MOI |
| Review Panel: | Immunology (82) |
III. Predicate Device
The predicate device IMMULITE® 2000 BR-MA, manufactured by Siemens Healthcare Diagnostics Products Ltd, Glyn Rhonwy, Llanberis, Wales, United Kingdom, was cleared by the FDA under K013984.
IV. Device Description
The IMMULITE® 2000 BR-MA assay was cleared under K013984. The components of the cleared assay were modified to reduce biotin interference.
The modified IMMULITE® 2000 BR-MA Assay is comprised of the following components:
| Component | Volume | Ingredients |
|---|---|---|
| BR-MA Bead Pack (L2BR12) | 200 beads | Polystyrene bead coated with Ligand-labeledanti-CA15-3 murine monoclonal antibody |
| BR-MA Reagent Wedge (L2BRA2) - Well 1 | 11.5 mL | Serum-based buffer, with preservative |
| BR-MA Reagent Wedge (L2BRA2) - Well 2 | 11.5 mL | Alkaline phosphatase (bovine calf intestine)conjugated to murine monoclonal anti-CA15-3antibody in buffer, with preservative |
| BR-MA Adjustors (LBRL, LBRH) | 3 mL each | Lyophilized CA15-3 in a nonhuman serum matrix,with preservative (low and high levels) |
The IMMULITE 2000 BR-MA is a solid-phase, two-step chemiluminescent immunometric assay. There are two incubation cycles of 30 minutes each.
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During the initial 30-minute cycle, the patient sample is incubated with biotinylated antibody coated bead (bead pack) and a buffer (reagent wedge well 1). The biotinylated antibody on the bead captures the antigen in the patient sample. On completion of the first 30-minute cycle, unbound sample/buffer are then removed via a centrifugal wash.
During the second 30-minute cycle, alkaline phosphatase antibody conjugate in buffer (reagent wedge well 2) is added to complete the bead pair immunocomplex sandwich consisting of capture Ab-antigendetection Ab. On completion of the second 30-minute cycle, unbound conjuqate is removed by centrifugal wash. The amount of alkaline phosphatase bound is directly proportional to the patient sample.
Following the two 30-minute incubation periods. IMMULITE chemiluminescent substrate (L2SUBM) is added for a further 5-minute incubation period to generate the luminogenic reaction.
The chemiluminescent substrate undergoes hydrolysis in the alkaline phosphatase to yield an unstable intermediate, which then emits photons. The sustained emissions are measured by the luminometer. The resulting relative light units are proportional to the concentration of CA15-3 in the sample, which is expressed as U/mL.
V. Intended Use / Indications for Use
For in vitro diagnostic use with the IMMULITE® 2000 Systems Analyzers - for the quantitative measurement of CA15-3 antigen in human serum and plasma, as an aid in the detection of recurrence in previously treated stage III breast cancer patients, and in the management of metastatic breast cancer patients by monitoring disease progression or response to treatment. Serial testing for patient CA15-3 values should be used in conjunction with other clinical methods used for detecting early recurrence in stage III disease and for monitoring response to treatment in patients with metastatic breast cancer.
VI. Comparison to Predicate Device
Comparison Table of Technological Characteristics
| Attribute | Candidate Device: IMMULITE® 2000 BR-MA Assay, modified | Predicate Device: IMMULITE® 2000 BR-MA Assay, K013984 |
|---|---|---|
| Intended Use / Indications for Use | For in vitro diagnostic use with theIMMULITE® 2000 Systems Analyzers – forthe quantitative measurement of CA15-3antigen in human serum and plasma, as anaid in the detection of recurrence inpreviously treated stage II and stage IIIbreast cancer patients, and in themanagement of metastatic breast cancerpatients by monitoring disease progressionor response to treatment. Serial testing forpatient CA15-3 values should be used inconjunction with other clinical methodsused for detecting early recurrence in stageII and stage III disease and for monitoringresponse to treatment in patients withmetastatic breast cancer. | Same |
| Analyte | Cancer Antigen 15-3 | Same |
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| Candidate Device:IMMULITE® 2000 BR-MA Assay, modified | Predicate Device:IMMULITE® 2000 BR-MA Assay,K013984 | |
|---|---|---|
| Attribute | IMMULITE® 2000 BR-MA Assay, modified | K013984 |
| Automated | Automated assay | Same |
| Measurement | Quantitative | Same |
| Sample Type | Human serum in plain tubes and BectonDickinson SST® vacutainer; heparinized andEDTA plasma | Same |
| Detection Limit | Detection Limits:Limit of Blank (LoB) = 0.21 U/mLLimit of Detection (LoD) = 0.30 U/mLLimit of Quantitation (LoQ) = 1 U/mLLoB, LoD, and LoQ were determined inaccordance with Clinical and LaboratoryStandards Institute (CLSI) EP17-A2 | Analytical Sensitivity: 1.0 U/mL |
| Assay MeasuringInterval | 1-300 U/mL | Same |
| Operating Principle | Immunologic sandwich | Same |
| Technology | Direct chemiluminescent | Same |
| Instrument | IMMULITE® 2000 Systems Analyzers | Same |
| Sample Volume | 5 μL | Same |
| Calibrator(Adjustors) | Lyophilized CA15-3 in a nonhuman serummatrix, with preservative | Same |
| Controls | Commercially available, minimum of 2 levels | Same |
| Detection Antibody | Alkaline phosphatase (bovine calfintestine) conjugated to murine monoclonalanti-CA15-3 antibody | Same |
| Capture Antibody | Anti-CA15-3 murine monoclonal antibody | Same |
| Biotin Interference | Specimens that contain biotin at aconcentration of 3500 ng/mL demonstrate aless than or equal to 10% change in results.Biotin concentrations greater than this maylead to incorrect results for patient samples. | Specimens that contain biotin at aconcentration of 100 ng/mLdemonstrate a less than or equalto 10% change in results. Biotinconcentrations greater than thismay lead to falsely depressedresults for patient samples |
VII. Summary of Performance Testing
Substantial equivalence of the modified IMMULITE® 2000 BR-MA assay was demonstrated by testing performance characteristics including detection capability, linearity, method comparison, precision, recovery, interference, hook effect, reference range and matrix comparison.
The following performance data are provided in support of a substantial equivalence determination.
i. Detection Limits
Limit of Blank (LoB), Limit of Detection (LoD), and Limit of Quantitation (LoQ) were determined in accordance with CLSI EP17-A2: Evaluation of Detection Capability for Clinical Laboratory Measurement Procedures; Approved Guideline – Second Edition. The LoB, LoD, and LoQ estimates are summarized below:
| LoB | 0.21 | U/mL |
|---|---|---|
| LoD | 0.30 | U/mL |
| LoQ | 1 | U/mL |
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ii. Measuring Interval / Linearity
Linearity studies were conducted with 3 reagent lots consistent with the governing standard CLSI EP06-ED2: Evaluation of the Linearity of Quantitative Measurement Procedures.
A high sample pool and low sample were mixed to prepare a linearity panel of ten levels. Linearity was determined if the allowable deviation from linearity (ADL) was ≤ 15% at each individual level.
Linearity was confirmed across the assay range by acceptable ADL at each individual level and supports the measuring interval of 1 - 300 U/mL.
iii. Method Comparison: Quantitative Assay
The method comparison study was performed comparing the modified device to the currently marketed device. A total of 274 patient samples covering the full range of the assay were analyzed. A single replicate was processed for each sample. Passing-Bablok regression was used to compare the devices.
| Lot | SpecimenType | Comparison Assay (x) | N | Regression Equation | Correlation Coefficient |
|---|---|---|---|---|---|
| 1 | Serum | IMMULITE 2000 BR-MA | 274 | $Y = 0.98x + 0.71$ | 0.989 |
| 2 | Serum | IMMULITE 2000 BR-MA | 274 | $Y = 0.99x + 0.16$ | 0.992 |
| 3 | Serum | IMMULITE 2000 BR-MA | 274 | $Y = 1.04x - 0.79$ | 0.990 |
iv. Verification of Assay Precision
Precision studies were conducted on one reagent lot on one IMMULITE 2000 analyzer in accordance with CLSI EP05-A3: Evaluation of Precision of Quantitative Measurement Procedures. Testing was performed on five serum samples spanning the range of the assay. Each sample was tested in duplicate over a period of 20 days, two runs per day, for a total of 40 runs and 80 replicates.
| Level | Mean | Within-Run | Total(Within-Lab) | ||
|---|---|---|---|---|---|
| (U/mL) | SD | %CV | SD | %CV | |
| 1 | 21.0 | 1.12 | 5.3 | 1.56 | 7.4 |
| 2 | 38.7 | 1.85 | 4.8 | 2.74 | 7.1 |
| 3 | 79.0 | 4.35 | 5.5 | 5.71 | 7.2 |
| 4 | 175 | 8.86 | 5.1 | 12.5 | 7.1 |
| 5 | 234 | 16.0 | 6.8 | 17.2 | 7.4 |
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v. Verification of Assay Reproducibility
Reproducibility studies were conducted on three reagents lot on one IMMULITE 2000 analyzer in accordance with CLSI EP05-A3: Evaluation of Precision of Quantitative Measurement Procedures, using the 5 x 5 x 3 experimental design. Testing was performed on five serum samples spanning the range of the assay. Each sample was tested over a period of five days, with five replicates per sample.
| Level | Mean(U/mL) | Total(Reproducibility) | |
|---|---|---|---|
| SD | %CV | ||
| 1 | 20.7 | 1.09 | 5.3 |
| 2 | 39.9 | 1.80 | 4.5 |
| 3 | 77.6 | 4.69 | 6.0 |
| 4 | 176 | 9.34 | 5.3 |
| 5 | 234 | 13.5 | 5.8 |
vi. Recovery
Spike and recovery studies were performed by spiking samples 1:19 (5% spike) with three CA15-3 solutions of differing concentrations.
| Sample | Neat SampleResult(U/mL) | SpikeSolution | Spiking solutionconcentration(U/mL) | ExpectedValue (U/mL) | ObservedValue (U/mL) | % Recovery |
|---|---|---|---|---|---|---|
| S1 | 48 | A | 360 | 64 | 61 | 95% |
| B | 780 | 85 | 86 | 101% | ||
| C | 1520 | 122 | 128 | 105% | ||
| S2 | 87 | A | 360 | 101 | 99 | 98% |
| B | 780 | 122 | 116 | 95% | ||
| C | 1520 | 159 | 161 | 101% | ||
| S3 | 102 | A | 360 | 115 | 115 | 100% |
| B | 780 | 136 | 125 | 92% | ||
| C | 1520 | 173 | 172 | 99% | ||
| S4 | 132 | A | 360 | 143 | 145 | 101% |
| B | 780 | 164 | 169 | 103% | ||
| C | 1520 | 201 | 208 | 103% | ||
| S5 | 222 | A | 360 | 229 | 240 | 105% |
| B | 780 | 250 | 233 | 93% | ||
| C | 1520 | 287 | 283 | 99% |
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vii. Interference
Verification of the assay interference was conducted in accordance with CLSI EP07-ED3: Interference Testing in Clinical Chemistry.
The following substances tested were determined to have no significant interference:
| Compound | InterferentConcentration |
|---|---|
| Hemoglobin | 381 mg/dL |
| Conjugated Bilirubin | 200 mg/L |
| Unconjugated Bilirubin | 200 mg/L |
| Intralipid | 3000 mg/dL |
| Biotin | 3500 ng/mL |
| 5-Fluorouracil | 1000 µg/mL |
| Cisplatin | 100 µg/mL |
| Cyclophosphamide | 1000 µg/mL |
| Doxorubicin Hydrochloride | 100 µg/mL |
| Mitomycin-C | 100 µg/mL |
| Vincristine | 1 µg/mL |
The following substances tested were found to have no detectable specificity (cross-reactivity):
| Compound | Cross-ReactantConcentration |
|---|---|
| Alpha-fetoprotein (AFP) | 5,000 IU/mL |
| Cancer Antigen 125 (CA125) | 10,000 U/mL |
| Cancer Antigen 19-9 (CA19-9) | 2,000 U/mL |
| Carcinoembryonic Antigen (CEA) | 5,000 ng/mL |
viii. Hook Effect
No hook effect was observed up to 80,000 U/mL. CA15-3 concentrations as high as 80,000 U/mL will report as >300 U/mL.
ix. Verification of Reference Range
The reference range was verified by assaying apparently healthy female samples according to CLSI EP28-A3C: Defining, Establishing, and Verifying Reference Intervals in the Clinical Laboratory. The existing reference range (6.4 - 58 U/mL) was verified.
| Lot 1 | Lot 2 | Lot 3 | |
|---|---|---|---|
| n Normal Females | 69 | 69 | 69 |
| n Normal Females samples between 6.4 - 58 U/mL | 65 | 65 | 65 |
| % Normal Females samples between 6.4 - 58 U/mL | 94% | 94% | 94% |
Siemens provides this information for reference. As with all in vitro diagnostic assays, each laboratory should determine its own reference ranges for the diagnostic evaluation of patient results. Consider these values as a guideline only.
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x. Matrix Comparison
The matrix comparison/specimen equivalence study was performed to evaluate the performance using different tube types (SST, Lithium Heparin, and EDTA). The study demonstrated comparable values to serum samples.
VIII. Conclusion
These performance studies support that the modified IMMULITE 2000 BR-MA Assay is substantially equivalent to the IMMULITE 2000 BR-MA Assay that is currently marketed, except for reduced biotin interference.
§ 866.6010 Tumor-associated antigen immunological test system.
(a)
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.(b)
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.