(626 days)
Not Found
No
The device description details a standard ELISA procedure and data analysis based on a calibration curve, with no mention of AI or ML techniques.
No
This device is an in vitro diagnostic (IVD) kit used for the quantification of free light chains to aid in the diagnosis of multiple myeloma and AL amyloidosis. It does not provide any treatment or therapeutic function.
Yes.
The device is intended for the quantification of free light chains, and this measurement "aids in the diagnosis of multiple myeloma and AL amyloidosis," which directly supports a diagnostic purpose.
No
The device description clearly outlines a physical kit involving reagents, microplates, washing steps, incubation, and spectrophotometry, indicating it is a hardware-based in vitro diagnostic device, not software-only.
Yes, this device is an IVD (In Vitro Diagnostic).
Here's why:
- Explicit Statement: The "Intended Use / Indications for Use" section clearly states: "For In Vitro Diagnostic Use."
- Intended Use: The device is intended for the quantification of substances (free light chains) in human serum, which is a biological sample taken from the body. This analysis is performed outside of the body ("in vitro").
- Diagnostic Purpose: The measurement of free light chains is stated to "aid in the diagnosis of multiple myeloma and AL amyloidosis." This indicates a diagnostic purpose.
- Laboratory Setting: The "Intended User / Care Setting" is listed as "laboratory / In Vitro Diagnostic Use," further confirming its use in a laboratory setting for diagnostic purposes.
- ELISA Procedure: The device utilizes an ELISA procedure, which is a common laboratory technique for in vitro diagnostic testing.
N/A
Intended Use / Indications for Use
The FLC Kappa kit is intended for the quantification of Kappa free light chains in human serum from adults with an Enzyme-Linked Immunosorbent Assay (ELISA) procedure. Measurement of free light chains aids in the diagnosis of multiple myeloma and AL amyloidosis. It must be used in conjunction with other laboratory and clinical findings. For In Vitro Diagnostic Use.
The FLC Lambda kit is intended for the quantification of Lambda free light chains in human serum from adults with an Enzyme-Linked Immunosorbent Assay (ELISA) procedure. Measurement of free light chains aids in the diagnosis of multiple myeloma and AL amyloidosis. It must be used in conjunction with other laboratory and clinical findings. For In Vitro Diagnostic Use.
Product codes (comma separated list FDA assigned to the subject device)
DFH, DEH
Device Description
The FLC Kappa and FLC Lambda test kits are intended for the quantification of free light chains in human serum from adults with an Enzyme-Linked Immunosorbent Assay (ELISA) procedure utilizing specific antibodies targeting anti-Lambda free light chains.
It is carried out in 8 successive steps:
- Incubation of the previously diluted samples and calibrators, in the wells of the microplate, where specific free light chain antibodies are fixed.
- Washing of the wells to remove elements that have not been fixed by the anti-free light chain antiserum.
- Incubation with an anti- light chain antiserum (Kit specific) conjugated to peroxidase. -
- Washing of the wells to remove the excess of antiserum conjugated to peroxidase. -
- Incubation with peroxidase substrate. -
- Stopping of the enzymatic reaction with an acidic solution. -
- -Reading of the optical density by absorbance spectrophotometry at 450 nm of the colored product.
- -Calculation of the free light chain concentration of the sample using a calibration curve obtained with calibrators that have been analyzed on the same microplate.
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
adults
Intended User / Care Setting
Not Found
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)
a) Precision and Reproducibility:
- Single-site reproducibility: Six different samples (4 serum, 2 controls) tested over 20 days with 3 replicates/sample, 2 runs/day, 1 reagent lot. Total 120 results per sample.
- FLC Kappa: Total reproducibility CV ranged from 10.4% to 14.8%.
- FLC Lambda: Total reproducibility CV ranged from 9.6% to 12.6%.
- Multi-operator reproducibility: Six different samples (4 serum, 2 controls) tested over 5 days by 3 operators, 5 replicates/sample, 1 reagent lot. Total 75 results per sample.
- FLC Kappa: Total reproducibility CV ranged from 7.4% to 12.9%.
- FLC Lambda: Total reproducibility CV ranged from 8.5% to 11.1%.
- Multi-lot reproducibility: Six different samples (4 serum, 2 controls) tested over 5 days by 1 operator, 5 replicates/sample, 3 reagent lots. Total 75 results per sample.
- FLC Kappa: Total reproducibility CV ranged from 6.9% to 9.4%.
- FLC Lambda: Total reproducibility CV ranged from 10.7% to 15.7%.
- Multi-site reproducibility: Six different samples (4 serum, 2 controls) tested over 5 days by 1 operator/site, 5 replicates/sample, 1 reagent lot across 3 laboratories.
- FLC Kappa: Total reproducibility CV ranged from 9.4% to 15.4%.
- FLC Lambda: Total reproducibility CV ranged from 11.4% to 15.2%.
b) Linearity/assay:
- Evaluated based on CLSI EP06-ed2: 2020 guideline. Three linearity panels from different patient sample pools.
- FLC Kappa: Linear range demonstrated between 4.5 mg/L and 76.2 mg/L at 1/1000 initial dilution.
- FLC Lambda: Linear range demonstrated between 3.8 mg/L and 66.8 mg/L at 1/1000 initial dilution.
c) Limit of Blank (LOB) / Limit of Detection (LOD) / Limit of Quantitation (LOQ):
- Evaluated based on CLSI EP17-A2 guideline.
- FLC Kappa: LOB = 0 mg/L, LOD = 0.8 mg/L, LOQ = 4.5 mg/L.
- FLC Lambda: LOB = 0 mg/L, LOD = 1.1 mg/L, LOQ = 3.8 mg/L.
d) Hook Effect (Antigen Excess): No interference observed.
e) Traceability: Calibration traceable to an internally assigned master calibrator.
f) Analytical specificity (Biological and Drug Interferences):
- Biological Interferences (conjugated bilirubin, unconjugated bilirubin, total protein, hemoglobin, triglycerides, rheumatoid factor) evaluated per CLSI EP7-A2 / EP07, 3rd Edition. No interference detected at tested concentrations.
- Drug Interferences (melphalan, dexamethazon, daratumumab, bortezomib, lenalidomide, pomalidomide, carfilzomib, isatuximab) evaluated per CLSI EP7-A2 / EP07, 3rd Edition. No interference detected at tested concentrations for FLC Lambda (table also indicates testing for Kappa).
g) Stability: Shelf-life and in-use studies conducted.
- Shelf Life: 9 months at 2 - 8 °C for both FLC Kappa and FLC Lambda.
- In-use: 5 uses within 1 month and 15 cumulative hours maximum at room temperature (19-25°C) for both FLC Kappa and FLC Lambda.
h) Expected values/ Reference range:
- Reference range intervals established using 238 healthy adults (US-population) per CLSI EP28-A3c.
- Kappa free light chain concentration: 6.4 - 17.4 mg/L
- Lambda free light chain concentration: 8.4 - 21.8 mg/L
- [Kappa free light chain] / [Lambda free light chain] ratio: 0.46 - 1.51
i) Comparison studies:
- Quantitative comparison: 216 serum samples for Kappa, 221 serum samples for Lambda, compared to predicate device using weighted Deming regression (CLSI EP09-A3).
- FLC Kappa: N=216, Sample Range 1.0-1947.0 mg/L, Slope = 0.557, Y-Intercept = 0.912, R2 = 0.917.
- FLC Lambda: N=221, Sample Range 1.6-860.4 mg/L, Slope = 0.608, Y-Intercept = 2.243, R2 = 0.749.
- Qualitative comparison: 216 serum samples for [Kappa FLC] / [Lambda FLC] ratio compared to a commercially available nephelometric technique.
- Positive Percent Agreement (PPA) = 82.3%
- Negative Percent Agreement (NPA) = 89.3%
j) Clinical Study (Diagnosis of multiple myeloma and AL amyloidosis):
- Total of 510 samples: 177 from multiple myeloma, 144 from AL amyloidosis, 189 non-myeloma/non-amyloidosis subjects.
- Clinical sensitivity for Multiple Myeloma (n=366) using [Kappa FLC] / [Lambda FLC] ratio: 96.6% (95% CI: 94.0% to 99.3%).
- Clinical specificity for Multiple Myeloma (n=366) using [Kappa FLC] / [Lambda FLC] ratio: 85.1% (95% CI: 79.4% to 89.5%).
- Clinical sensitivity for AL Amyloidosis (n=333) using [Kappa FLC] / [Lambda FLC] ratio: 91.0% (95% CI: 86.3% to 95.7%).
- Clinical specificity for AL Amyloidosis (n=333) using [Kappa FLC] / [Lambda FLC] ratio: 85.1% (95% CI: 79.4% to 89.5%).
Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
Clinical sensitivity: 96.6% (95% confidence interval: 94.0% to 99.3%) for Multiple Myeloma.
Clinical specificity: 85.1% (95% confidence interval: 79.4% to 89.5%) for Multiple Myeloma.
Clinical sensitivity: 91.0% (95% confidence interval: 86.3% to 95.7%) for AL Amyloidosis.
Clinical specificity: 85.1% (95% confidence interval: 79.4% to 89.5%) for AL Amyloidosis.
Positive Percent Agreement (%): 82.3
Negative Percent Agreement (%): 89.3
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
§ 866.5550 Immunoglobulin (light chain specific) immunological test system.
(a)
Identification. An immunoglobulin (light chain specific) immunological test system is a device that consists of the reagents used to measure by immunochemical techniques both kappa and lambda types of light chain portions of immunoglobulin molecules in serum, other body fluids, and tissues. In some disease states, an excess of light chains are produced by the antibody-forming cells. These free light chains, unassociated with gamma globulin molecules, can be found in a patient's body fluids and tissues. Measurement of the various amounts of the different types of light chains aids in the diagnosis of multiple myeloma (cancer of antibody-forming cells), lymphocytic neoplasms (cancer of lymphoid tissue), Waldenstrom's macroglobulinemia (increased production of large immunoglobulins), and connective tissue diseases such as rheumatoid arthritis or systemic lupus erythematosus.(b)
Classification. Class II (performance standards).
0
Image /page/0/Picture/0 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health and Human Services logo. To the right of that is the FDA logo, which is a blue square with the letters "FDA" in white. To the right of the blue square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.
November 18, 2022
Sebia, Inc. Karen Anderson Director of Regulatory 1705 Corporate Drive, Suite 400 Norcross, Georgia 30093
Re: K210623
Trade/Device Name: FLC Kappa, FLC Lambda Regulation Number: 21 CFR 866.5550 Regulation Name: Immunoglobulin (Light Chain Specific) Immunological Test System Regulatory Class: Class II Product Code: DFH, DEH Dated: August 2, 2022 Received: August 4, 2022
Dear Karen Anderson:
We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database located at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.
If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.
1
Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-products); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.
Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.
For comprehensive regulatory information about 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
Enclosure
2
Indications for Use
510(k) Number (if known) K210623
Device Name FLC Kappa FLC Lambda
Indications for Use (Describe)
The FLC Kappa kit is intended for the quantification of Kappa free light chains in human serum from adults with an Enzyme-Linked Immunosorbent Assay (ELISA) procedure. Measurement of free light chains aids in the diagnosis of multiple myeloma and AL amyloidosis. It must be used in conjunction with other laboratory and clinical findings. For In Vitro Diagnostic Use.
The FLC Lambda kit is intended for the quantification of Lambda free light chains in human serum from adults with an Enzyme-Linked Immunosorbent Assay (ELISA) procedure. Measurement of free light chains aids in the diagnosis of multiple myeloma and AL amyloidosis. It must be used in conjunction with other laboratory and clinical findings. For In Vitro Diagnostic Use.
| 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
510K SUMMARY (Summary of Safety and Effectiveness)
This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of 21 CFR 807.92.
| Submitter Name | Sebia, Inc. |
|---|---|
| Address | 1705 Corporate Drive Suite 400 |
| Norcross, Georgia 30093, USA | |
| Contact | Karen Anderson, Dir of Regulatory, Sebia Inc. |
| Phone: 1-800-835-6497 | |
| Fax: 770-446-8511 | |
| Email: karen.anderson@sebia-usa.com | |
| Matthew C Wagner, Scientific Affairs Specialist | |
| Phone 1-800-835-6497, 3752 | |
| Fax 770-446-8511 | |
| Email: Matthew.Wagner@sebia-usa.com | |
| Date Prepared | October 20, 2022 |
| Manufacturing | Sebia |
| Parc Technologique Léonard de Vinci | |
| Rue Léonard de Vinci, | |
| CP 8010 LISSES, 91008 EVRY Cedex | |
| FRANCE | |
| Phone: (33) 1 69 89 80 80 | |
| Fax: (33) 1 69 89 78 78 | |
| Product Name | FLC Kappa |
| FLC Lambda | |
| Common Name | Light Chain immunological test system |
| Product Regulation No. | 21CFR sec. 866.5550 - Immunoglobulin (light chain |
| specific) immunological test system | |
| 21 CFR sec. 862.1660- Quality Control Material | |
| (assayed and unassayed) | |
| Product Codes | DFH, Kappa antigen, antiserum, control |
| DEH, Lambda, antigen, antiserum, control | |
| Device classification and Panel | |
| Classification | Class II, Immunology (82) |
| Establishment Registration No. | 8023024 |
4
1. DEVICE DESCRIPTIONS
The FLC Kappa and FLC Lambda test kits are intended for the quantification of free light chains in human serum from adults with an Enzyme-Linked Immunosorbent Assay (ELISA) procedure utilizing specific antibodies targeting anti-Lambda free light chains.
It is carried out in 8 successive steps:
- Incubation of the previously diluted samples and calibrators, in the wells of the । microplate, where specific free light chain antibodies are fixed.
- Washing of the wells to remove elements that have not been fixed by the anti-free light chain antiserum.
- Incubation with an anti- light chain antiserum (Kit specific) conjugated to peroxidase. -
- Washing of the wells to remove the excess of antiserum conjugated to peroxidase. -
- Incubation with peroxidase substrate. -
- Stopping of the enzymatic reaction with an acidic solution. -
- -Reading of the optical density by absorbance spectrophotometry at 450 nm of the colored product.
- -Calculation of the free light chain concentration of the sample using a calibration curve obtained with calibrators that have been analyzed on the same microplate.
2. REAGENTS
REAGENTS AND MATERIALS SUPPLIED IN THE FLC KAPPA AND FLC LAMBDA KITS
| PN 5102 | PN 5103 | |
|---|---|---|
| ITEMS | FLC Kappa Kit | FLC Lambda Kit |
| Microplate Kappa 1 plate with 12 segments, 8 wells each | Microplate Kappa 1 plate with 12 segments, 8 wells each | Microplate Lambda 1 plate with 12 segments, 8 wells each |
| Dilution buffer (ready to use) | 1 vial, 100 mL | 1 vial, 100 mL |
| Wash solution (stock solution) | 1 vial, 100 mL | 1 vial, 100 mL |
| Calibrators (ready to use) | 5 vials, 0.6 mL each | |
| Kappa Calibrators | 5 vials, 0.6 mL each | |
| Lambda Calibrators | ||
| PER antiserum (ready to use) | 1 vial, 12 mL | |
| Anti-Kappa - PER antiserum (ready to use) | 1 vial, 12 mL | |
| Anti-Lambda - PER antiserum (ready to use) | ||
| Substrate | 1 vial, 12 mL | |
| Substrate Kappa (ready to use) | 1 vial, 12 mL | |
| Substrate Lambda (ready to use) | ||
| Stop solution (ready to use) | 1 vial, 12 mL | 1 vial, 12 mL |
5
REAGENTS/SUPPLIES REQUIRED BUT NOT SUPPLIED IN THE KITS
| SUPPLIES | SEBIA PRODUCT NUMBER |
|---|---|
| Densitometer for microplate reading by | |
| absorbance spectrophotometry at 450 nm. | |
| NON COATED ELISA PLATES (10), SEBIA, | |
| microplates with scored wells in order to | |
| complete segments when the forecasted | |
| number of samples per segment is fewer than | |
| 8, and frame | |
| to store the segments that have not been | |
| used. | PN 5303 |
| Absorbent paper for removal the excess of | |
| wash solution from the wells of the microplate. | |
| Multichannel pipette. | |
| FLC CONTROL LEVEL 1 | PN 5112 |
| FLC CONTROL LEVEL 2 | PN 5113 |
3. INDICATIONS FOR USE
FLC Kappa and FLC Lambda kits
The FLC Kappa kit is intended for the quantification of Kappa free light chains in human serum from adults with an Enzyme-Linked Immunosorbent Assay (ELISA) procedure. Measurement of free light chains aids in the diagnosis of multiple myeloma and AL amyloidosis. It must be used in conjunction with other laboratory and clinical findings.
For In Vitro Diagnostic Use only.
The FLC Lambda kit is intended for the quantification of Lambda free light chains in human serum from adults with an Enzyme-Linked Immunosorbent Assay (ELISA) procedure. Measurement of free light chains aids in the diagnosis of multiple myeloma and AL amyloidosis. It must be used in conjunction with other laboratory and clinical findings. For In Vitro Diagnostic Use only.
Special conditions for use statements: For prescription use only.
Warning: The result of the FLC Kappa and FLC Lambda in a given specimen determined with assays and/or instrument platforms from different manufacturers can vary due to differences in assay methods and reagent
specificity. The results reported by the laboratory to the physician must include the identity of the assay used. Values obtained with different assay methods cannot be used interchangeably.
6
SUBSTANTIAL EQUIVALENCE INFORMATION
| Predicate Device Name | Predicate Device
510(k) number | Regulation No. |
|--------------------------------------------------------------------------------------|-----------------------------------|----------------|
| The Binding Site Freelite®
Human Kappa Free Kit for use on
the Siemens BN™ II | K031016 | 866.5550 |
| The Binding Site Freelite®
Human Lambda Free Kit for use
on the Siemens BN™ II | K031016 | 866.5550 |
4. COMPARISON WITH PREDICATE DEVICE
Kit Similarities (Table A).
| Similarities | ||
|---|---|---|
| Candidate Device | Predicate | |
| Table A | FLC Kappa | |
| FLC Lambda | The Binding Site Freelite® Human Kappa Free | |
| and Freelite® Human Lambda Free kits for use | ||
| on the Siemens BN TM II (K031016) | ||
| Analyte | Kappa: Kappa FLC | |
| Lambda: Lambda FLC | Same | |
| Measurement | Quantitative | Same |
Kit Differences (Table B)
| Differences | ||
|---|---|---|
| Table B | Candidate Device | Predicate Device |
| Indications for Use | The FLC Kappa kit is intended for the | |
| quantification of Kappa free light | ||
| chains in human serum from adults | ||
| with an Enzyme-Linked | ||
| Immunosorbent Assay (ELISA) | ||
| procedure. Measurement of free light | ||
| chains aids in the diagnosis of multiple | ||
| myeloma and AL amyloidosis. It must | ||
| be used in conjunction with other | ||
| laboratory and clinical findings. | ||
| For In Vitro Diagnostic Use only. | Kappa: This kit is intended for the | |
| quantitation of kappa free light chains in | ||
| serum and urine on the Siemens BN™ II. | ||
| Measurement of free light chains aids in the | ||
| diagnosis and monitoring of multiple | ||
| myeloma, lymphocytic neoplasms, | ||
| Waldenstrom's macroglobulinemia, AL | ||
| amyloidosis, light chain deposition disease | ||
| and connective tissue diseases such as | ||
| systemic lupus erythematosus in | ||
| conjunction with other laboratory and clinical | ||
| findings. | ||
| The FLC Lambda kit is intended for the | ||
| quantification of Lambda free light | ||
| chains in human serum from adults | ||
| with an Enzyme-Linked | ||
| Immunosorbent Assay (ELISA) | ||
| procedure. Measurement of free light | ||
| chains aids in the diagnosis of multiple | ||
| myeloma and AL amyloidosis. It must | ||
| be used in conjunction with other | ||
| laboratory and clinical findings. | ||
| For In Vitro Diagnostic Use only. | Lambda: This kit is intended for the | |
| quantitation of lambda free light chains in | ||
| serum and urine on the Siemens BN™ II | ||
| Measurement of free light chains aids in the | ||
| diagnosis and monitoring of multiple | ||
| myeloma, lymphocytic neoplasms, | ||
| Waldenstrom's macroglobulinemia, AL | ||
| amyloidosis, light chain deposition disease | ||
| and connective tissue diseases such as | ||
| systemic lupus erythematosus in | ||
| conjunction with other laboratory and clinical | ||
| findings. | ||
| Specimen | ||
| Type | Human Serum | Human Serum, Human Urine |
| Detection | ||
| Method | Enzyme-Linked Immunosorbent | |
| Assay (ELISA) | Nephelometric | |
| Detection | ||
| Antibody | Kappa : Sandwich ELISA with | |
| polyclonal rabbit anti- human | ||
| kappa free light chains coated | ||
| on the well of the microplate | ||
| (capture antibody) and | ||
| polyclonal rabbit anti-human | ||
| kappa light chain conjugated to | ||
| horseradish peroxidase (HRP) | ||
| (detection antibody) |
Lambda : Sandwich ELISA with
polyclonal rabbit anti-human lambda
free light chain coated on the well of
the microplate (capture antibody) and
polyclonal rabbit anti-human lambda
light chain conjugated to horseradish
peroxidase (HRP) (detection antibody) | Kappa: Polyclonal sheep anti-human kappa
antibody coated with latex particles
Lambda: Polyclonal sheep anti-human
Lambda antibody coated with latex particles |
| Analytical
Measuring
ranges | Kappa
Standard dilution (1/1000)
4.5 to 76.2 mg/L
(dilution scheme 1/250 to 1/100 000)
Lambda
Standard dilution (1/1000)
3.8 to 66.8 mg/L
(dilution scheme 1/250 to 1/100 000) | Kappa
Standard dilution (1/100)
5.9 to 190 mg/L
(dilution scheme 1/5 to 1/8000)
Lambda
Standard dilution (1/100)
5.0 to 160 mg/L
(dilution scheme 1/5 to 1/8000) |
| Reference
Interval | Kappa: 6.4 to 17.4 mg/L.
Lambda: 8.4 to 21.8 mg/L.
Ratio: 0.46 to 1.51 | Kappa: 3.30 to 19.40 mg/L
Lambda: 5.71 to 26.30 mg/L
Ratio: 0.26-1.65 |
| Similarities | | |
| Item | Candidate | Predicate |
| Reference Material | Internal preparation | Reference Same |
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8
Calibrators:
| Differences | ||
|---|---|---|
| Item | Candidate | Predicate |
| Number of levels | 5 | One |
5. PERFORMANCE DATA
a) Precision and Reproducibility
The precision and FLC Kappa and FLC Lambda assays were evaluated according to the CLSI EP05-A3 guideline.
Single-site reproducibility
Six different samples were tested using the FLC Kappa and FLC Lambda kits. The analyzed samples included 4 serum samples S1 to S4) and 2 controls (samples C1 and C2). Each day, for 20 days, a unique operator analyzed the same samples (3 replicates / sample) on a microplate (2 runs with minimum 2 hours between the 2 runs) with 1 reagent lot (same microplate design each day), yielding a total of 120 results per sample.
The single-site reproducibility study is summarized in the following table including within-run, between runs, within-day, between-days and total reproducibility precision estimates (SD and %CV) for the free light chain concentrations (in mg/L).
| Sample | Mean
(mg/L) | Within-run | | Between-runs | | Within-day | | Between-days | | Total
(*)
reproducibility
(Within-laboratory
precision) | |
|--------|----------------|------------|------|--------------|-------|------------|-------|--------------|------|---------------------------------------------------------------------|-------|
| | | SD | CV | SD | CV | SD | CV | SD | CV | SD | CV |
| S1 | 39.2 | 1.47 | 3.8% | 4.02 | 10.2% | 4.28 | 10.9% | 1.29 | 3.3% | 4.47 | 11.4% |
| S2 | 6.3 | 0.41 | 6.6% | 0.74 | 11.9% | 0.85 | 13.6% | 0.36 | 5.8% | 0.92 | 14.8% |
| S3 | 14.4 | 0.54 | 3.7% | 1.30 | 9.0% | 1.40 | 9.7% | 0.81 | 5.6% | 1.62 | 11.2% |
| S4 | 62.0 | 2.89 | 4.7% | 6.03 | 9.7% | 6.69 | 10.8% | 1.77 | 2.9% | 6.92 | 11.2% |
| C1 | 11.5 | 0.67 | 5.8% | 1.09 | 9.5% | 1.28 | 11.2% | 0.00 | 0.0% | 1.28 | 11.2% |
| C2 | 42.2 | 1.64 | 3.9% | 4.00 | 9.5% | 4.32 | 10.2% | 0.66 | 1.6% | 4.37 | 10.4% |
FLC Kappa
(*) Total reproducibility includes within-run, between-runs, within-day and between-days.
9
FLC Lambda
| Sample | Mean
(mg/L) | Within-run | | Between-runs | | Within-day | | Between-days | | Total
reproducibility
(Within-laboratory
precision)(*) | |
|--------|----------------|------------|------|--------------|------|------------|------|--------------|------|-----------------------------------------------------------------|-------|
| | | SD | CV | SD | CV | SD | CV | SD | CV | SD | CV |
| S1 | 33.7 | 1.55 | 4.6% | 2.67 | 7.9% | 3.09 | 9.2% | 1.61 | 4.8% | 3.48 | 10.3% |
| S2 | 13.2 | 0.55 | 4.2% | 0.96 | 7.3% | 1.11 | 8.4% | 0.63 | 4.8% | 1.28 | 9.7% |
| S3 | 20.0 | 0.83 | 4.1% | 1.25 | 6.2% | 1.50 | 7.5% | 0.74 | 3.7% | 1.67 | 8.3% |
| S4 | 79.5 | 5.13 | 6.5% | 5.15 | 6.5% | 7.27 | 9.1% | 6.91 | 8.7% | 10.03 | 12.6% |
| C1 | 19.7 | 1.33 | 6.8% | 1.40 | 7.1% | 1.93 | 9.8% | 0.00 | 0.0% | 1.93 | 9.8% |
| C2 | 37.1 | 1.66 | 4.5% | 2.76 | 7.5% | 3.22 | 8.7% | 1.55 | 4.2% | 3.57 | 9.6% |
(*) Total reproducibility includes within-run, between-runs, within-day and between-days.
Multi-operator reproducibility
Six different samples were tested using the FLC Kappa and FLC Lambda kits. The analyzed samples included 4 serum samples (samples S1 to S4) and 2 controls (samples C1 and C2). Each day, for 5 days, 3 operators analyzed the same samples (5 replicates / sample) on a microplate, with 1 reagent lot (same microplate design each day), yielding a total of 75 results per sample. The multi-operator reproducibility study is summarized in the following within-day, between-days, within-operator, between-operators and total reproducibility precision estimates (SD and %CV) for the free light chain concentrations (in mg/L).
| Sample | Mean
(mg/L) | Within-day | | Between-days | | Within-operator | | Between-
operators | | Total
reproducibility (*) | |
|--------|----------------|------------|------|--------------|------|-----------------|-------|-----------------------|------|------------------------------|-------|
| | | SD | CV | SD | CV | SD | CV | SD | CV | SD | CV |
| S1 | 44.1 | 2.49 | 5.7% | 2.05 | 4.7% | 3.23 | 7.3% | 0.50 | 1.1% | 3.27 | 7.4% |
| S2 | 6.8 | 0.46 | 6.8% | 0.63 | 9.2% | 0.78 | 11.4% | 0.00 | 0.0% | 0.78 | 11.4% |
| S3 | 15.7 | 0.72 | 4.6% | 1.30 | 8.3% | 1.49 | 9.5% | 0.00 | 0.0% | 1.49 | 9.5% |
| S4 | 67.9 | 3.52 | 5.2% | 5.58 | 8.2% | 6.60 | 9.7% | 0.00 | 0.0% | 6.60 | 9.7% |
| C1 | 12.7 | 1.25 | 9.9% | 0.93 | 7.3% | 1.56 | 12.3% | 0.50 | 3.9% | 1.64 | 12.9% |
| C2 | 44.8 | 1.95 | 4.3% | 2.94 | 6.6% | 3.53 | 7.9% | 0.00 | 0.0% | 3.53 | 7.9% |
FLC Kappa
(*) Total reproducibility includes within-day, between-days, within-operator and between-operators.
FLC Lambda
| Sample | Mean
(mg/L) | Within-day | | Between-days | | Within-operator | | Between-operators | | Total
reproducibility (*) | |
|--------|----------------|------------|------|--------------|------|-----------------|-------|-------------------|------|------------------------------|-------|
| | | SD | CV | SD | CV | SD | CV | SD | CV | SD | CV |
| S1 | 35.4 | 2.70 | 7.6% | 2.87 | 8.1% | 3.94 | 11.1% | 0.00 | 0.0% | 3.94 | 11.1% |
| S2 | 13.4 | 0.87 | 6.5% | 0.83 | 6.2% | 1.20 | 9.0% | 0.00 | 0.0% | 1.20 | 9.0% |
| S3 | 21.1 | 0.98 | 4.6% | 1.49 | 7.1% | 1.79 | 8.5% | 0.00 | 0.0% | 1.79 | 8.5% |
| S4 | 81.5 | 5.88 | 7.2% | 5.58 | 6.8% | 8.11 | 10.0% | 3.99 | 4.9% | 9.03 | 11.1% |
| C1 | 20.7 | 1.59 | 7.7% | 1.41 | 6.8% | 2.13 | 10.3% | 0.00 | 0.0% | 2.13 | 10.3% |
| C2 | 37.1 | 2.09 | 5.6% | 2.81 | 7.6% | 3.51 | 9.4% | 1.93 | 5.2% | 4.00 | 10.8% |
(*) Total reproducibility includes within-day, between-days, within-operator and between-operators.
10
Multi-lot reproducibility
Six different samples were tested using the FLC Kappa and FLC Lambda kits. The analyzed samples included 4 serum samples $1 to S4), and 2 controls (samples C1 and C2), Each day, for 5 days, 1 operator analyzed the same samples (5 replicates / sample) on a microplate, with 3 reagent lots (1 microplate per reagent lot, same microplate design each day), yielding a total of 75 results per sample. The multi-lot reproducibility study is summarized in the following table including between-days, within day, between-lots, within-lot and total reproducibility precision estimates (SD and %CV) for the free light chain concentrations (in mg/L).
FLC Kappa
| Sample | Mean
(mg/L) | Between-days | | Within-day | | Between-lots | | Within-lot | | Total reproducibility
(*) | |
|--------|----------------|--------------|------|------------|------|--------------|------|------------|------|------------------------------|------|
| | | SD | CV | SD | CV | SD | CV | SD | CV | SD | CV |
| S1 | 38.7 | 1.72 | 4.4% | 2.00 | 5.2% | 0.76 | 2.0% | 2.64 | 6.8% | 2.75 | 7.1% |
| S2 | 6.8 | 0.50 | 7.4% | 0.36 | 5.3% | 0.17 | 2.6% | 0.62 | 9.1% | 0.64 | 9.4% |
| S3 | 15.3 | 0.55 | 3.6% | 0.50 | 3.3% | 0.40 | 2.6% | 0.74 | 4.9% | 0.84 | 5.5% |
| S4 | 62.2 | 1.69 | 2.7% | 3.59 | 5.8% | 1.65 | 2.6% | 3.96 | 6.4% | 4.29 | 6.9% |
| C1 | 12.1 | 0.54 | 4.4% | 0.86 | 7.1% | 0.00 | 0.0% | 1.01 | 8.3% | 1.01 | 8.3% |
| C2 | 42.8 | 1.71 | 4.0% | 1.96 | 4.6% | 1.44 | 3.4% | 2.60 | 6.1% | 2.97 | 6.9% |
(*) Total reproducibility includes between-days, within-day, between-lots and within-lot.
FLC Lambda
| Sample | Mean
(mg/L) | Between-days | | Within-day | | Between-lots | | Within-lot | | Total
reproducibility (*) | |
|--------|----------------|--------------|-------|------------|------|--------------|-------|------------|-------|------------------------------|-------|
| | | SD | CV | SD | CV | SD | CV | SD | CV | SD | CV |
| S1 | 33.7 | 3.12 | 9.3% | 2.91 | 8.6% | 0.70 | 2.1% | 4.27 | 12.7% | 4.33 | 12.9% |
| S2 | 13.8 | 1.20 | 8.7% | 1.14 | 8.3% | 1.40 | 10.1% | 1.66 | 12.0% | 2.17 | 15.7% |
| S3 | 20.8 | 1.98 | 9.5% | 0.99 | 4.8% | 2.20 | 10.6% | 2.21 | 10.7% | 3.12 | 15.0% |
| S4 | 71.4 | 7.63 | 10.7% | 5.71 | 8.0% | 0.00 | 0.0% | 9.53 | 13.4% | 9.53 | 13.4% |
| C1 | 20.9 | 1.51 | 7.2% | 1.70 | 8.2% | 2.32 | 11.1% | 2.27 | 10.9% | 3.25 | 15.6% |
| C2 | 38.5 | 3.29 | 8.5% | 1.84 | 4.8% | 1.66 | 4.3% | 3.77 | 9.8% | 4.12 | 10.7% |
(*) Total reproducibility includes between-days, within-day, between-lots and within-lot.
Multi-site reproducibility
Six different samples were tested using the FLC Kappa and the FLC Lambda kits. The analyzed samples included 4 serum samples (samples S1 to S4) and 2 controls (samples C1 and C2). Each day, for 5 days, 1 operator analyzed the same samples (1 dilution 1/1000, 5 replicates / sample) on a microplate with 1 reagent lot (same microplate design each day). The same protocol was followed by 2 other operators in 2 other laboratories with the same reagent lot.
The multi-sites reproducibility study is summarized in the following within-day, between-days, within-site, between-sites and total reproducibility precision estimates (SD and %CV) for the free light chain concentrations (in mg/L).
11
FLC Kappa
| Sample | Mean
(mg/L) | Between-days | | Within-day | | Between-sites | | Within-site | | Total reproducibility
(*) | |
|--------|----------------|--------------|------|------------|------|---------------|-------|-------------|-------|------------------------------|-------|
| | | SD | CV | SD | CV | SD | CV | SD | CV | SD | CV |
| S1 | 6.2 | 0.39 | 6.3% | 0.45 | 7.2% | 0.00 | 0.0% | 0.60 | 9.6% | 0.60 | 9.6% |
| S2 | 15.0 | 1.48 | 9.9% | 0.87 | 5.8% | 0.00 | 0.0% | 1.71 | 11.4% | 1.71 | 11.4% |
| S3 | 32.5 | 2.76 | 8.5% | 1.29 | 4.0% | 1.48 | 4.5% | 3.05 | 9.4% | 3.38 | 10.4% |
| S4 | 71.9 | 5.78 | 8.0% | 4.23 | 5.9% | 2.86 | 4.0% | 7.16 | 9.9% | 7.71 | 10.7% |
| C1 | 13.8 | 0.70 | 5.0% | 1.08 | 7.9% | 1.69 | 12.2% | 1.29 | 9.3% | 2.12 | 15.4% |
| C2 | 57.2 | 2.68 | 4.7% | 2.67 | 4.7% | 3.79 | 6.6% | 3.78 | 6.6% | 5.36 | 9.4% |
(*) Total reproducibility includes between-days, within-day, between-sites and within-site.
FLC Lambda
| Sample | Mean (mg/L) | Between-days | Within-day | Between-sites | Within-site | Total reproducibility (*) | |||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| SD | CV | SD | CV | SD | CV | SD | CV | SD | CV | ||
| S1 | 11,4 | 0,91 | 8,0% | 0,76 | 6,7% | 0,51 | 4,5% | 1,19 | 10,5% | 1,30 | 11,4% |
| S2 | 20,7 | 1,62 | 7,8% | 1,51 | 7,3% | 1,12 | 5,4% | 2,22 | 10,7% | 2,48 | 12,0% |
| S3 | 31,3 | 3,39 | 10,8% | 2,12 | 6,8% | 1,87 | 6,0% | 4,00 | 12,8% | 4,42 | 14,1% |
| S4 | 68,8 | 6,13 | 8,9% | 4,44 | 6,5% | 4,81 | 7,0% | 7,57 | 11,0% | 8,97 | 13,0% |
| C1 | 24,2 | 2,27 | 9,4% | 1,62 | 6,7% | 2,40 | 9,9% | 2,78 | 11,5% | 3,68 | 15,2% |
| C2 | 50,1 | 5,81 | 11,6% | 3,87 | 7,7% | 2,83 | 5,7% | 6,98 | 13,9% | 7,53 | 15,1% |
(*) Total reproducibility includes between-days, within-day, between-sites and within-site.
b) Linearity/assay
The linearity of the FLC Kappa and FLC Lambda kits was evaluated in a study based on the Clinical and Laboratory Standards Institute (CLSI - USA) EP06-ed2: 2020 guideline "Evaluation of the Linearity of Quantitative Measurement Procedures".
The results for concentration (mg/L) of Kappa and Lambda free light chains were analyzed using statistical tools recommended by CLSI.
Three linearity panels at 1/1000 dilution were performed based on 3 different patient sample pools.
| FLC Assay | Range
Tested
(mg/L) | Linear
Regression | 95% CI Slope | 95%
CI
Y-Intercept | Claimed
linear
range |
|------------------|---------------------------|----------------------|------------------|--------------------------|----------------------------|
| Kappa study N°1 | [4.4; 81.7] | Y=1.009x -0.2967 | [0.9534; 1.065] | [-0.88; 0.29] | |
| Kappa study N°2 | [4.5; 79.9] | Y=1.016x -0.2014 | [0.967; 1.065] | [-0.72; 0.32] | [4.5mg/L;
76.2mg/L] |
| Kappa study N°3 | [3.5; 76.2] | $Y=0.9332x + 0.4952$ | [0.8635; 1,003] | [-0.11; 1.10] | |
| Lambda study N°1 | [3.8; 74.1] | Y=1.046x -1.537 | [0.9389; 1.153] | [-2.52; -0.55] | |
| Lambda study N°2 | [3.3; 77.7] | Y=0.9561x -0.62 | [0.9299; 0.9823] | [-0.84; -0.40] | [3.8mg/L;
66.8mg/L] |
| Lambda study N°3 | [3.5; 66.8] | Y=0.9611x -0.31 | [0.8948; 1.027] | [-0.87; 0.25] | |
12
The highest of the lower limit and the lowest of the higher limit found in the 3 linearity panels were taken.
The linearity of FLC Kappa kit at initial dilution 1/1000 is demonstrated between 4.5 mg/L and 76.2 ma/L.
The linearity of FLC Lambda kit at initial dilution 1/1000 is demonstrated between 3.8 mg/L and 66.8 mq/L.
c) Limit of Blank (LOB) / Limit of Detection (LOD) / Limit of Quantitation (LOQ)
The determination of the limit of detection (LOD) and of the limit of quantitation (LOQ) of the FLC Kappa kit was evaluated in a study based on the Clinical and Laboratory Standards Institute (CLSI - USA) EP17-A2 guideline "Evaluation of Detection Capability for Clinical Laboratory Measurement Procedures; Approved Guideline - Second Edition".
Due to the blank removal, the limit of blank (LOB) is considered as being 0 mg/L.
The LOD was determined by assaying 5 samples with low Kappa free light chains concentration: they were tested in two runs of four replicates over the course of seven days using two reagent lots (a total of 56 replicates per sample for each lot). The precision profile analysis was used to calculate the LOD. Since LOD values for the 2 lots are below the calibrator 1 values, the LOD value is considered as equal to the LOQ value.
The LOQ was determined by assaying 5 samples with low Kappa free light chains concentration: they were tested in two runs of four replicates over the course of seven days using two reagent lots (a total of 56 replicates per sample for each lot). The LoQ was defined to be the lowest concentration level that meets the within-laboratory imprecision of