(223 days)
Not Found
No
The summary describes a standard immunoassay kit and its performance characteristics. There is no mention of AI or ML in the device description, intended use, or performance studies.
No
Explanation: This device is an in vitro diagnostic immunoassay used for the semi-quantitative determination of thyroid stimulating autoantibodies to aid in the diagnosis of Graves' disease. It does not provide any treatment or therapeutic benefit.
Yes
The IMMULITE® 2000 TSI Assay is an in vitro diagnostic immunoassay that aids in the diagnosis of patients suspected of having Graves' disease.
No
The device description clearly lists physical components like "TSI bead pack," "TSI reagent wedge," "TSI adjustors," "TSI controls," and "Multi-Diluent 2," indicating it is an in vitro diagnostic assay kit with tangible reagents and materials, not solely software.
Yes, this device is an IVD (In Vitro Diagnostic).
The intended use explicitly states: "The IMMULITE® 2000 TSI (thyroid-stimulating immunoglobulins) Assay is an in vitro diagnostic immunoassay..." and "The IMMULITE® TSI Calibration Verification Material (CVM) is for in vitro diagnostic use...".
The device description also details components used for laboratory testing of human samples (serum and plasma).
N/A
Intended Use / Indications for Use
The IMMULITE® 2000 TSI (thyroid-stimulating immunoglobulins) Assay is an in vitro diagnostic immunoassay for the semi-quantitative determination of thyroid stimulating autoantibodies specific to thyroid stimulating hormone receptors (TSHR) in human serum (including Serum Separator tubes) or plasma (K2-EDTA or lithium heparin). The IMMULITE® 2000 TSI Assay is for use on the IMMULITE® 2000 system. The measurement of thyroid stimulating autoantibodies, in conjunction with other clinical and laboratory findings, is used as an aid in the diagnosis of patients suspected of having Graves' disease.
The IMMULITE® TSI Calibration Verification Material (CVM) is for in vitro diagnostic use in the verification of calibration of the IMMULITE® TSI Assay on the IMMULITE® 2000 Systems.
Product codes (comma separated list FDA assigned to the subject device)
JZO
Device Description
The IMMULITE 2000 TSI assay kit consists of the following components:
- TSI bead pack coated with MAb (3D7) anti-TSHR anchor antibody and hTSHR Capture Chimera
- TSI reagent wedge containing hTSHR-Chimera alkaline phosphatase conjugate
- TSI adjustors: low and high, containing TSI negative heat-inactivated bovine serum and ● thyroid stimulating human MAb (M22)
- TSI controls: negative, low, and high, containing TSI negative human serum and thyroid stimulating MAb (M22)
- Multi-Diluent 2 ●
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
Not Found
Indicated Patient Age Range
Not Found
Intended User / Care Setting
For prescription use only.
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)
Precision/Reproducibility:
Study Type: Precision evaluation based on CLSI document EP05-A2.
Sample Size: Six serum samples and low/high kit controls. Each sample tested in duplicate in two runs per day over 20 days, totaling 80 replicates.
Key Results:
| Sample | Mean IU/L | SD (Repeatability) | %CV (Repeatability) | SD (Within Lab) | %CV (Within Lab) |
|---|---|---|---|---|---|
| L2SIC1 | 1.04 | 0.05 | 4.9 | 0.06 | 5.4 |
| L2SIC2 | 21.27 | 0.96 | 4.5 | 1.21 | 5.7 |
| SERP1 | 0.34 | 0.02 | 7.0 | 0.03 | 8.3 |
| SERP2 | 0.69 | 0.03 | 4.1 | 0.03 | 5.0 |
| SERP3 | 1.57 | 0.07 | 4.4 | 0.08 | 5.3 |
| SERP4 | 4.43 | 0.18 | 4.0 | 0.26 | 5.9 |
| SERP5 | 7.80 | 0.27 | 3.5 | 0.42 | 5.4 |
| SERP6 | 29.09 | 1.91 | 6.6 | 2.11 | 7.3 |
Linearity/Assay Reportable Range:
Study Type: Linearity evaluation according to CLSI EP6-A.
Sample Size: Gravimetrically pooled low and high TSI positive and negative samples from commercial vendors.
Key Results: Linearity data for % Difference was shown to be ± 15% or 0.50 IU/L (whichever is greater) for most samples; tested within range 0.50 - 40.0 IU/L with calibrators and pooled samples. Certain patient samples may exhibit non-linear dilution due to the heterogeneity of TSHR autoantibodies.
Traceability, Stability, Expected Values:
Traceability: Traceable to NIBSC standard 08/204.
Assay and Component Stability:
Real time shelf life stability testing for unopened devices and opened components.
Acceptance criteria based on analyte drift as described in CLSI document EP25-A.
Closed Vial Testing: 0, 60, 120, 180, 270, 360, 450, and 540 days. Four replicates of Adjustors, Controls, MDP, and Calibrator J.
Open Vial/Kit Testing: 0, 7, 8, 14, 30, 45, 60, 90, and 97 days (2-8°C).
Open Vial/Aliquotted Components Testing: 0, 30, 60, 90, 120, 150, 180, and 194 days (-20°C).
Key Results: Supports stability claims as listed in "Component" table in the original text (e.g., Kit, unopened: 12 Months at 2 – 8 °C).
Detection Limit:
Limit of Blank (LoB): Determined non-parametrically. Five TSI negative samples analyzed on three IMMULITE® 2000 instruments over 3 days, using three reagent lots, 2 runs with 2 replicates per run. Total 180 blank measurements.
Key Results: Highest LoB was 0.03 IU/L.
Limit of Detection (LoD): Five TSI serum samples (mean concentrations from 0.016 to 0.110 IU/L) assayed in replicates of 4 using 3 reagent kit lots run for 3 days on 3 systems. 36 observations per sample.
Key Results: Highest LoD was 0.06 IU/L.
Analytical Specificity:
Endogenous interferents: Intralipid (500 and 1000 mg/dL); Hemoglobin (200 and 500 mg/dL); Bilirubin: Conjugated and Unconjugated (20 and 40 mg/dL each); K2-EDTA (9mg/mL); Heterophile samples, HAMA and Rheumatoid Factor.
Key Results: No interference (≤ 10% different than control sample) except for hemoglobin (presence at 200mg/dL or higher may affect recovery). Short Draw K2-EDTA Plasma samples may result in under-recovery.
HAMA Study: Bias
§ 866.5870 Thyroid autoantibody immunological test system.
(a)
Identification. A thyroid autoantibody immunological test system is a device that consists of the reagents used to measure by immunochemical techniques the thyroid autoantibodies (antibodies produced against the body's own tissues). Measurement of thyroid autoantibodies may aid in the diagnosis of certain thyroid disorders, such as Hashimoto's disease (chronic lymphocytic thyroiditis), nontoxic goiter (enlargement of thyroid gland), Grave's disease (enlargement of the thyroid gland with protrusion of the eyeballs), and cancer of the thyroid.(b)
Classification. Class II (performance standards).
0
Image /page/0/Picture/1 description: The image shows the logo for the Department of Health & Human Services - USA. The logo consists of a circular seal with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" arranged around the perimeter. Inside the circle is a stylized image of three human profiles facing to the right, with flowing lines representing hair or movement.
March 3, 2016
Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002
Siemens Healthcare Diagnostics, Inc. Susan Brocchi Regulatory Clinical Affairs Specialist 511 Benedict Avenue Tarrytown NY 10591
Re: K152061
Trade/Device Name: Immulite 200 TSI Assay, Immulite 200 TSI Calibration Verification Material
Regulation Number: 21 CFR 866.5870 Regulation Name: Thyroid autoantibody immunological test system Regulatory Class: II Product Code: JZO Dated: February 3, 2016 Received: February 4, 2016
Dear Ms. Brocchi:
We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. 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.
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 Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
1
If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638 2041 or (301) 796-7100 or at its Internet address
http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to
http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.
You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address
http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.
Sincerely yours,
Kelly OlinerAS
For.
Leonthena R. Carrington, MS, MBA, MT(ASCP) Director Division of Immunology and Hematology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health
Enclosure
2
Indications for Use
510(k) Number (if known) K152061
Device Name IMMULITE®2000 TSI Assay
IMMULITE®2000 TSI Calibration Verification Material (CVM)
Indications for Use (Describe)
The IMMULITE® 2000 TSI (thyroid-stimulating immunoglobulins) Assay is an in vitro diagnostic immunoassay for the semi-quantitative determination of thyroid stimulating autoantibodies specific to thyroid stimulating hormone receptors (TSHR) in human serum (including Serum Separator tubes) or plasma (K2-EDTA or lithium heparin). The IMMULITE® 2000 TSI Assay is for use on the IMMULITE® 2000 system. The measurement of thyroid stimulating autoantibodies, in conjunction with other clinical and laboratory findings, is used as an aid in the diagnosis of patients suspected of having Graves' disease.
The IMMULITE® TSI Calibration Material (CVM) is for in vitro diagnostic use in the verification of calibration of the IMMULITE® TSI Assay on the IMMULITE® 2000 Systems.
X Prescription Use (Part 21 CFR 801 Subpart D)
Over-The-Counter Use (21 CFR 801 Subpart C)
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3
IMMULITE® 2000 TSI Assay
510(k) Summary as Required by 21 CFR 807.92
A. 510(k) Number:
- B. Purpose for Submission: New Device
- C. Measurand: Thyroid-stimulating immunoglobulins
- D. Type of test: Semi-quantitative chemiluminescent immunometric assay
- E. Applicant: Siemens Laboratory Diagnostics Inc.
- F. Proprietary and Established Names: IMMULITE®2000 TSI Assay IMMULITE®2000 TSI Calibration Verification Material
- G. Regulatory Information:
| Regulation Section | Classification | Product Code | Panel |
|---|---|---|---|
| 21 CFR 866.5870, | |||
| Thyroid autoantibody | |||
| immunological test | |||
| system | Class 2 (Performance | ||
| Standards) | JZO – System, Test, | ||
| Thyroid Autoantibody | Immunology (82) | ||
| 21 CFR 862.1150 | Class 2 | JIT, Calibrator, | |
| secondary | Clinical Chemistry | ||
| (75) | |||
| 21 CFR 862.1660, | |||
| Quality control | |||
| material (assayed and | |||
| unassayed) | Class 1, reserved | JJX, Single analyte | |
| control | Clinical Chemistry | ||
| (75) |
H. Intended Use:
-
- Intended use(s):
The IMMULITE® 2000 TSI (thyroid-stimulating immunoglobulins) Assay is an in vitro diagnostic immunoassay for the semi-quantitative determination of thyroid stimulating autoantibodies specific to thyroid stimulating hormone receptors (TSHR) in human serum (including Serum Separator tubes) or plasma (K2-EDTA or lithium heparin). The IMMULITE® 2000 TSI Assay is for use on the IMMULITE® 2000 system. The measurement of thyroid stimulating autoantibodies, in conjunction with other clinical and laboratory findings, is used as an aid in the diagnosis of patients suspected of having Graves' disease.
- Intended use(s):
The IMMULITE® TSI Calibration Verification Material (CVM) is for in vitro diagnostic
4
use in the verification of calibration of the IMMULITE® TSI Assay on the IMMULITE® 2000 Systems.
-
- Indication(s) for use: See Intended Use above
-
- Special conditions for use statement(s): For prescription use only
-
- Special instrument requirements: Only for use on the IMMULITE 2000 Analyzer (K970227)
I. Device Description:
The IMMULITE 2000 TSI assay kit consists of the following components:
- TSI bead pack coated with MAb (3D7) anti-TSHR anchor antibody and hTSHR Capture Chimera
- TSI reagent wedge containing hTSHR-Chimera alkaline phosphatase conjugate
- TSI adjustors: low and high, containing TSI negative heat-inactivated bovine serum and ● thyroid stimulating human MAb (M22)
- TSI controls: negative, low, and high, containing TSI negative human serum and thyroid stimulating MAb (M22)
- Multi-Diluent 2 ●
J. Substantial Equivalence Information:
-
- Comparison with predicate:
| ASSAY SIMILARITIES – Predicate 1 | ||
|---|---|---|
| Item | Device | Predicate 1 |
| IMMULITE 2000 TSI Assay | Thyretain TSI Reporter BioAssay | |
| Analyte | TSH receptor stimulating | |
| autoantibodies | same | |
| Signal | Chemiluminescence | same |
| Detection | ||
| method | Luminometry | same |
| Controls | 3 (negative, low, and high) | 3 (negative, reference, positive) |
| ASSAY DIFFERENCES – Predicate 1 | ||
| Item | Device | Predicate |
| IMMULITE® 2000 TSI Assay | Thyretain TSI Reporter BioAssay |
5
| Intended Use | The IMMULITE® 2000 TSI
(thyroid-stimulating
immunoglobulins) Assay is an in
vitro diagnostic immunoassay for
the semi-quantitative determination
of thyroid stimulating autoantibodies
specific to thyroid stimulating
hormone receptors (TSHR) in
human serum (including Serum
Separator tubes) or plasma (K2-
EDTA or lithium heparin). The
IMMULITE® 2000 TSI Assay is for
use on the IMMULITE® 2000
system. The measurement of thyroid
stimulating autoantibodies, in
conjunction with other clinical and
laboratory findings, is used as an aid
in the diagnosis of patients
suspected of having Graves' disease. | Intended for the qualitative detection
in serum of thyroid stimulating
autoantibodies to the thyroid
stimulating hormone receptors
(TSHRs) on the thyroid. The detection
of these stimulating autoantibodies, in
conjunction with other clinical and
laboratory findings, may be useful as
an aid in the differential diagnosis of
patients with Graves' disease. |
|--------------------|------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Assay Principal | Immunometric | Cell-based Bioassay |
| Sample matrix | Serum or plasma | Serum |
| Assay format | Semi-quantitative | Qualitative |
| Unit of measure | IU/L | SRR% (sample to reference ratio) |
| Solid phase | Polystyrene bead coated with
hTSHR capture chimera | CHO Mc4 cell monolayer |
| Traceability | NIBSC Standard 08/204 | NISBC Standard 03/192 |
| Sample size | 50 uL | 40 uL |
| Measuring
range | 0.10-40 IU/L | N/A (qualitative) |
| Incubation time | 60 minutes | 3 hours |
| LoB | 0.03 IU/L | 62.75% |
| LoD | 0.06 IU/L | 89.14% |
| Assay Cut-off | 0.55 IU/L | 140% |
| ASSAY SIMILARITIES – Predicate 2 | ||
|---|---|---|
| Item | Device | Predicate 2 |
| IMMULITE 2000 TSI Assay | Elecsys Anti-TSHR Immunoassay | |
| Analyte | TSH receptor stimulating | |
| autoantibodies | Same | |
| Unit of | ||
| measure | IU/L | Same |
| Sample size | 50μL | Same |
| ASSAY DIFFERENCES – Predicate 2 | ||
| Item | Device | Predicate |
| IMMULITE® 2000 TSI Assay | Elecsys Anti-TSHR Immunoassay | |
| Intended Use | The IMMULITE® 2000 TSI (thyroid-stimulating immunoglobulins) Assay is an in vitro diagnostic immunoassay for the semi-quantitative determination of thyroid stimulating autoantibodies specific to thyroid stimulating hormone receptors (TSHR) in human serum (including Serum Separator tubes) or plasma (K2-EDTA or lithium heparin). The IMMULITE® 2000 TSI Assay is for use on the IMMULITE® 2000 system. The measurement of thyroid stimulating autoantibodies, in conjunction with other clinical and laboratory findings, is used as an aid in the diagnosis of patients suspected of having Graves' disease. | An immunoassay for the in vitro quantitative determination of autoantibodies to TSH receptor in human serum using a human thyroid stimulating monoclonal antibody. The anti-TSH receptor determination is used in the assessment of patients with suspect Graves' disease (autoimmune hyperthyroidism). The electrochemiluminescence immunoassay “ECLIA” is intended for use on Elecsys and cobas e immunoassay analyzers. |
| Sample matrix | Serum or plasma | Serum |
| Controls | 3 (negative, low, and high) | 2 (positive and negative) |
| Detecting | ||
| Antibody | hTSHR AP detection Chimera | M22 |
| Assay Protocol | Bridge Format | Competition Principle |
| Detection | ||
| Protocol | Chemiluminescence | Electrochemiluminescence |
| Traceability | NIBSC Standard 08/204 | NISBC Standard 90/672 |
| Incubation | 60 Minutes | 27 Minutes |
| LoB | 0.03 IU/L | 0.500 IU/L |
| LoD | 0.06 IU/L | 0.800 IU/L |
| LoQ | 0.10 IU/L | 0.900 IU/L |
| Reportable | ||
| Range | 0.10-40 IU/L | 0.800 - 40.0 IU/L |
| Assay Cut-off | 0.55 IU/L | 1.75 IU/L |
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7
| Calibrator Verification Materials | ||
|---|---|---|
| SIMILARITIES | ||
| Item | Device | Predicate |
| IMMULITE® 2000 TSI CVM | Elecsys Anti-TSHR CalCheck | |
| Form | Lyophilized | Same |
| Stability | Stable unopened until the expiration date. | Same |
| Use | Single Use Only | Same |
| Analyte | Thyroid Stimulating IgG | Same |
| Storage | 2-8°C | Same |
| Calibrator Verification Materials | ||
| DIFFERENCES | ||
| Item | Device | Predicate |
| IMMULITE® 2000 TSI CVM | Elecsys Anti-TSHR CalCheck | |
| Intended Use | The IMMULITE® TSI Calibration Verification Material (CVM) is for in vitro diagnostic use in the verification of calibration of the IMMULITE® TSI Assay on the IMMULITE® 2000 Systems. | For use in the verification of calibration established by the Elecsys Anti-TSHR reagent on the indicated Elecsys and cobas e immunoassay analyzers. |
| Instrumentation | Siemens IMMULITE® 2000 | Roche Elecsys and cobas e |
| Sample Matrix | Bovine serum with preservatives | Human serum |
K. Standard/Guidance Documents Referenced:
CLSI EP05-A2, Evaluation of Precision Performance of Quantitative Measurement Methods, Approved Guideline – Second Edition
CLSI EP06-A, Evaluation of Linearity of Quantitative Measurement Methods: A Statistical Approach – First Edition
CLSI EP07-A2, Interference Testing in Clinical Chemistry; Approved Guideline - Second Edition
8
CLSI EP09-A3, Measurement Procedure Comparison and Bias Estimation Using Patient Samples - Third Edition
CLSI EP17-A2, Evaluation of Detection Capability for Clinical Laboratory Measurement Procedures: Approved Guideline - Second Edition
CLSI EP24-A2, Assessment of the Diagnostic Accuracy of Laboratory Tests Using Receiver Operating Characteristic Curves; Approved Guideline - Second Edition
CLSI EP25-A, Evaluation of Stability of in vitro Diagnostic Reagents; Approved Guidance – First Edition
CLSI EP28-A3c, Defining, Establishing, and Verifying Reference Intervals in the Clinical Laboratory – Third Edition
L. Test Principle:
The IMMULITE 2000 TSI assay is an automated, two-cycle, chemiluminescent immunoassay. It employs two recombinant chimeric human TSH receptors (hTSHR) where the major epitope for the blocking antibody is replaced. The capture receptor is immobilized on the solid phase (polystyrene bead) by 3D7, a monoclonal antibody directed against the C-terminus of hTSHR. In the first 30-minute cvcle, thyroid stimulating antibodies in the patient sample bind through one arm to the capture receptor on the polystyrene bead. Upon completion of the 1st cycle, the bead is washed 4X with water onboard the instrument. In the second cycle, a liquid reagent containing the signal receptor is added to the reaction tube and incubated for 30-minutes. The immobilized TSI binds the signal receptor through the second arm forming a bridge. Unbound signal receptor is then removed by four centrifugal water washes. Finally, chemiluminescent substrate is added to the reaction tube and a signal is generated in direct relation to the amount of bound signal receptor and TSI in the sample.
M. Performance Characteristics
-
- Analytical Performance:
- a. Precision/Reproducibility:
Precision was evaluated using a protocol based on CLSI document EP05-A2. Six serum samples and the low and high kit controls were tested. Serum samples were prepared by mixing positive samples into negative sample pools in order to reach the desired concentration.
Each sample was tested in duplicate in two runs per day over 20 days, for a total of 80 replicates. One instrument was used, and one lot of the device was tested for a total of 80 replicates per sample. Precision was calculated using Nested Analysis of Variance.
9
| 20-Day
Imprecision
| N=80 | Repeatability | Within Lab | |||
|---|---|---|---|---|---|
| Sample | Mean | ||||
| IU/L | SD | %CV | SD | %CV | |
| L2SIC1 | 1.04 | 0.05 | 4.9 | 0.06 | 5.4 |
| L2SIC2 | 21.27 | 0.96 | 4.5 | 1.21 | 5.7 |
| SERP1 | 0.34 | 0.02 | 7.0 | 0.03 | 8.3 |
| SERP2 | 0.69 | 0.03 | 4.1 | 0.03 | 5.0 |
| SERP3 | 1.57 | 0.07 | 4.4 | 0.08 | 5.3 |
| SERP4 | 4.43 | 0.18 | 4.0 | 0.26 | 5.9 |
| SERP5 | 7.80 | 0.27 | 3.5 | 0.42 | 5.4 |
| SERP6 | 29.09 | 1.91 | 6.6 | 2.11 | 7.3 |
b. Linearity/assay Reportable Range:
IMMULITE 2000 TSI Linearity was evaluated according to CLSI EP6-A. TSI positive and TSI negative samples were obtained from several commercial vendors. Sample sets were prepared gravimetrically pooling low and high samples.
Conclusion
Linearity data for % Difference was shown to be ± 15% or 0.50 IU/L (whichever is greater) for most samples; tested within range 0.50 - 40.0 IU/L with calibrators and pooled samples. Because of the heterogeneity of TSHR autoantibodies, certain patient samples may exhibit a non-linear dilution.
c. Traceability, Stability, Expected Values (controls, calibrators, methods):
Traceability:
The IMMULITE® 2000 TSI assay is traceable to NIBSC standard 08/204 and is manufactured using qualified materials and measurement procedures.
The instrument-stored assay calibration curve is based on a series of calibrators covering the claimed assay range and is specific for each lot of the assay.
Assay and component stability:
Real time shelf life stability testing to support the following claims for the unopened devices and opened components:
10
Acceptance criteria for all components of the assay was assessed based on analyte drift as described in CLSI document EP25-A. The dose value of the component should be within the limits described in the table below except if below the LoQ of 0.10 IU/L where recovery cannot be properly determined. At each time point, all Control results must be within Quality Control established range, to validate the run.
| Component on Stability | Parameter | Drift Specifications |
|---|---|---|
| Reagents and Beads | Control 1 at ≤ -60°C | ≤ 15% from Day 0 mean |
| Reagents and Beads | Control 2 at ≤ -60°C | ≤ 10% from Day 0 mean |
| Reagents and Beads | CVM 1 at ≤ -60°C | ≤ 0.15 IU/L from Day 0 mean |
| Reagents and Beads | MDP 1 | ≤ 20% from Day 0 mean |
| Reagents and Beads | MDP 4 | ≤ 10% from Day 0 mean |
| Reagents and Beads | Cal J | ≤ 10% from Day 0 mean |
Closed vial testing (kit stored at 2 - 8 °C) was conducted at 0, 60, 120, 180, 270, 360, 450, and 540 days. Four replicates for the Low and High Adjustors and four replicates of each Control, MDP, and Calibrator J were run on each day of the study. Dose values of the Controls and samples were calculated from the Day 0 adjustment and from the adjustment performed on each subsequent day of testing.
Open vial/kit testing of components stored at 2 - 8°C was conducted at Days 0, 7, 8, 14, 30, 45, 60, 90, and 97. Open vial testing of aliquotted components stored at -20°C was conducted on Days 0, 30, 60, 90, 120, 150, 180, and 194. Four replicates of each Adjustor and four replicates of each Control were run on each day of the study.
| Component | Storage | Stability Claim |
|---|---|---|
| Kit, unopened | 2 – 8 °C | 12 Months |
| Bead Pack, open | 2 – 8 °C | 90 Days |
| Reagent wedge, open and on-board | 2 – 8 °C | 90 Days |
| Sample diluent, open (L2M2Z, L2M2Z4) | 2 – 8 °C | 30 Days |
| Sample diluent, open frozen aliquoted (L2M2Z, L2M2Z4) | -20 °C | 6 Months |
| Adjustors open (L2SIL, L2SIH) | 2 – 8 °C | 90 Days |
| Adjustors (L2SIL, L2SIH) frozen aliquotted | -20 °C | 4 Months |
| Controls open (L2SIC1, L2SIC2) | 2 – 8 °C | 90 Days |
| Controls (L2SIC1, L2SIC2) frozen aliquotted | -20 °C | 6 Months |
| CVM, unopened | 2 – 8 °C | 12 Months |
| CVM, opened and reconstituted | 2 – 8 °C | 30 Days |
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Sample stability:
The sample stability study supports the claim of stability for 24 hours at 20 - 25°C, 7 days at 2 - 8°C, and 12 months at -20°C for serum and plasma.
Adjustment interval (calibration curve) stability:
Real time testing supported the recommended adjustment interval of four weeks.
- d. Detection Limit:
Limit of Blank (LoB): Five TSI negative samples were analyzed on three IMMULITE® 2000 instruments over 3 days. The samples were tested using three lots of reagent for 2 runs with 2 replicates per run. The LoB result was determined by applying a nonparametric principle based on ranked ordered value using the following equation where p= (100-a) = 95 and NB = number of blank measurements = 180:
LoB rank position = [NB(p/100)+0.5]
The LoB was the 171.5th ranked result. The LoB was analyzed for each of the 3 lots. The highest LoB by lot was determined to be 0.03 IU/L.
Limit of Detection (LoD): Five TSI serum samples (mean concentrations ranging from 0.016 to 0.110 IU/L) were assayed in replicates of 4 using 3 reagent kit lots run for 3 days on 3 systems. Thirty-six observations were obtained for each sample. The LoD was determined by the following equation:
$$\mathbf{LoD} = \mathbf{LoB} + \mathbf{c}_p \mathbf{SD_L}$$
The LoD was analyzed for each of the 3 lots. The highest LoD by lot was determined to be 0.06 IU/L.
- e. Analytical Specificity:
Endogenous interferents:
Three serum pools with concentrations across the assay range were prepared and spiked separately with endogenous interferents. A control sample was prepared for each interfering substance by spiking the appropriate diluent at the same volume as the interfering substance; each sample was tested in triplicate. The interferents tested were: Intralipid (500 and 1000 mg/dL); Hemoglobin (200 and 500 mg/dL); Bilirubin: Conjugated and Unconjugated (20 and 40 mg/dL each); and K2-EDTA (9mg/mL). Heterophile samples, HAMA and Rheumatoid Factor, were also evaluated.
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No interference (≤ 10% different than control sample) was found when the means of control samples were compared to the means of the spiked samples, except in the case of hemoglobin. The sponsor has included limitations to this effect in the package insert: "Presence of hemoglobin at a concentration of 200mg/dL or higher may affect recovery."
Samples spiked with a five-fold excess of K2-EDTA anticoagulant were compared to normal serum to investigate the effect of a short blood draw on TSI assay results. Three samples were tested; all but one showed recovery within ± 10% of the serum control value. The sponsor has included limitations to this effect in the package insert: "Short Draw K2-EDTA Plasma samples may result in under-recovery of IMMULITE 2000 TSI results."
HAMA Study:
TSI samples were prepared at three target concentrations across the assay range. Samples were divided into control groups (spiked with HAMA negative serum) and test groups (spiked with HAMA positive serum to final HAMA concentration of 10 ng/mL or 40 ng/mL). Mean values for each sample were determined on IMMULITE 2000 (n=12) and the percent (%) difference from the control samples was calculated.
The assay demonstrated individual sample bias of