(90 days)
Not Found
No
The device description details a standard immunometric assay with chemiluminescent detection, and there is no mention of AI or ML in the intended use, device description, or performance studies.
No
This device is for in vitro diagnostic use, meaning it analyzes samples (serum and plasma) to measure CA15-3 antigen. It aids in detecting cancer recurrence or monitoring disease progression, but it does not directly treat or provide therapy to a patient.
Yes
The "Intended Use / Indications for Use" states "For in vitro diagnostic use" and describes its use "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." This clearly indicates its role in diagnosing and monitoring a disease.
No
The device description clearly outlines physical components (Bead Pack, Reagent Wedge, Adjustors) and a chemical process (solid-phase, two-step chemiluminescent immunometric assay) performed on an analyzer (IMMULITE® 2000 Systems Analyzers). This indicates it is a hardware-based in vitro diagnostic assay, not a software-only device.
Yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Intended Use/Indications for Use: The very first sentence explicitly states "For in vitro diagnostic use". It also describes the measurement of an antigen (CA15-3) in human serum and plasma, which are biological samples taken from the body for analysis outside of the body.
- Device Description: The description details a laboratory assay that uses chemical reactions and measurements (chemiluminescent immunometric assay, relative light units) to determine the concentration of a substance (CA15-3) in a sample. This is characteristic of an in vitro diagnostic test.
- Performance Studies: The performance studies described (detection capability, linearity, method comparison, precision, etc.) are standard evaluations for IVD devices to demonstrate their analytical performance.
The entire description aligns with the definition and characteristics of an In Vitro Diagnostic device.
N/A
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.
Product codes
MOI
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:
- BR-MA Bead Pack (L2BR12): 200 beads, Polystyrene bead coated with Ligand-labeled anti-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-CA15-3 antibody 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.
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.
Mentions image processing
Not Found
Mentions AI, DNN, or ML
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Input Imaging Modality
Not Found
Anatomical Site
Not Found
Indicated Patient Age Range
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Intended User / Care Setting
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Description of the training set, sample size, data source, and annotation protocol
Not Found
Description of the test set, sample size, data source, and annotation protocol
Not Found
Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
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.
-
Detection Limits:
- Study Type: Evaluation of Detection Capability for Clinical Laboratory Measurement Procedures; Approved Guideline – Second Edition (CLSI EP17-A2).
- Key Results: Limit of Blank (LoB) = 0.21 U/mL, Limit of Detection (LoD) = 0.30 U/mL, Limit of Quantitation (LoQ) = 1 U/mL.
-
Measuring Interval / Linearity:
- Study Type: Linearity studies conducted consistent with CLSI EP06-ED2: Evaluation of the Linearity of Quantitative Measurement Procedures.
- Sample Size: Linearity panel of ten levels created from a high sample pool and low sample.
- Key Results: Linearity was confirmed across the assay range (1 - 300 U/mL) with allowable deviation from linearity (ADL) ≤ 15% at each individual level.
-
Method Comparison: Quantitative Assay:
- Study Type: Method comparison against the currently marketed predicate device.
- Sample Size: 274 patient samples covering the full range of the assay.
- Key Results: Passing-Bablok regression with correlation coefficients:
- Lot 1: 0.989 ($Y = 0.98x + 0.71$)
- Lot 2: 0.992 ($Y = 0.99x + 0.16$)
- Lot 3: 0.990 ($Y = 1.04x - 0.79$)
-
Verification of Assay Precision:
- Study Type: Precision studies conducted in accordance with CLSI EP05-A3: Evaluation of Precision of Quantitative Measurement Procedures.
- Sample Size: Five serum samples spanning the range of the assay. Each sample tested in duplicate over 20 days, two runs/day, total 40 runs and 80 replicates.
- Key Results: Total (Within-Lab) %CV ranged from 7.1% to 7.4%.
-
Verification of Assay Reproducibility:
- Study Type: Reproducibility studies conducted in accordance with CLSI EP05-A3: Evaluation of Precision of Quantitative Measurement Procedures, using the 5 x 5 x 3 experimental design.
- Sample Size: Five serum samples spanning the range of the assay. Each sample tested over five days, with five replicates per sample.
- Key Results: Total (Reproducibility) %CV ranged from 4.5% to 6.0%.
-
Recovery:
- Study Type: Spike and recovery studies.
- Key Results: Percent recovery ranged from 92% to 105%.
-
Interference:
- Study Type: Verification of assay interference conducted in accordance with CLSI EP07-ED3: Interference Testing in Clinical Chemistry.
- Key Results: No significant interference from tested substances (Hemoglobin, Bilirubin, Intralipid, Biotin up to 3500 ng/mL, various chemotherapy drugs). No detectable cross-reactivity from AFP, CA125, CA19-9, CEA.
-
Hook Effect:
- Key Results: No hook effect observed up to 80,000 U/mL. Concentrations as high as 80,000 U/mL will report as >300 U/mL.
-
Verification of Reference Range:
- Study Type: Verified by assaying apparently healthy female samples according to CLSI EP28-A3C: Defining, Establishing, and Verifying Reference Intervals in the Clinical Laboratory.
- Sample Size: 69 normal females.
- Key Results: 94% of normal female samples were within the existing reference range (6.4 - 58 U/mL).
-
Matrix Comparison:
- Study Type: Specimen equivalence study.
- Key Results: Demonstrated comparable values to serum samples when using different tube types (SST, Lithium Heparin, and EDTA).
Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
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Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.
Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.
Not Found
Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).
Not Found
§ 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.
0
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"
1
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
2
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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3
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 514 | |
| Newark, 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-labeled |
| anti-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-CA15-3 | ||
| antibody 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.
4
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 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 II and 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 | ||
| II and stage III disease and for monitoring | ||
| response to treatment in patients with | ||
| metastatic breast cancer. | Same | |
| Analyte | Cancer Antigen 15-3 | Same |
5
| | 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 Becton
Dickinson SST® vacutainer; heparinized and
EDTA plasma | Same |
| Detection Limit | Detection Limits:
Limit of Blank (LoB) = 0.21 U/mL
Limit of Detection (LoD) = 0.30 U/mL
Limit of Quantitation (LoQ) = 1 U/mL
LoB, LoD, and LoQ were determined in
accordance with Clinical and Laboratory
Standards Institute (CLSI) EP17-A2 | Analytical Sensitivity: 1.0 U/mL |
| Assay Measuring
Interval | 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 serum
matrix, with preservative | Same |
| Controls | Commercially available, minimum of 2 levels | Same |
| Detection Antibody | Alkaline phosphatase (bovine calf
intestine) conjugated to murine monoclonal
anti-CA15-3 antibody | Same |
| Capture Antibody | Anti-CA15-3 murine monoclonal antibody | Same |
| Biotin Interference | Specimens that contain biotin at a
concentration of 3500 ng/mL demonstrate a
less than or equal to 10% change in results.
Biotin concentrations greater than this may
lead to incorrect results for patient samples. | Specimens that contain biotin at a
concentration of 100 ng/mL
demonstrate a less than or equal
to 10% change in results. Biotin
concentrations greater than this
may lead to falsely depressed
results 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 | Specimen
Type | 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 Sample
Result
(U/mL) | Spike
Solution | Spiking solution
concentration
(U/mL) | Expected
Value (U/mL) | Observed
Value (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 | Interferent
Concentration |
|---------------------------|------------------------------|
| 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-Reactant
Concentration |
|--------------------------------|---------------------------------|
| 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.
9
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.