(120 days)
The Elecsys Vitamin D total II assay is intended for the quantitative determination of total 25-hydroxyvitamin D in human serum and plasma. This assay is to be used as an aid in the assessment of vitamin D sufficiency in adults. The electrochemiluminescence binding assay is intended for use on the cobas e 411 immunoassay analyzer.
CalSet Vitamin D total II is used for calibrating the quantitative Elecsys Vitamin D total II assay on the cobas e 411 immunoassay analyzer.
PreciControl Vitamin D total II is used for quality control of the Elecsys Vitamin D total II assay on the cobas e 411 immunoassay analyzer.
This CalCheck set is an assayed control for use in calibration verification of the assay range established by the Elecsys Vitamin D total II reagent on the cobas e 411 immunoassay analyzer.
Elecsys Vitamin D total II is a second generation assay by Roche Diagnostics for the in vitro quantitative determination of 25-hydroxyvitamin D in human serum and plasma. It is intended for use on the cobas e 411 immunoassay analyzer. The cobas e family of analyzers employs the electrochemiluminescence "ECLIA" technology. The assay is a 27 minute assay utilizing a competition principle and a pretreatment step to release the bound 25-hydroxyvitamin D from the vitamin D binding protein.
Results are determined via a calibration curve which is instrument specifically generated by 2-point calibration against the master curve for that reagent lot.
The provided text describes the acceptance criteria and study proving the device meets those criteria for the Elecsys Vitamin D total II assay and its associated calibrators and controls.
Here's the breakdown of the information requested:
1. Table of Acceptance Criteria and Reported Device Performance
The document does not explicitly present a "table of acceptance criteria" in terms of specific thresholds for performance metrics. Instead, it describes various studies conducted to demonstrate performance and states that the data supports substantial equivalence. The "reported device performance" is woven throughout Section 4, "Non-Clinical Performance Evaluation," and Section 5, "External (Clinical) Testing."
However, we can infer some performance metrics and the studies that support them. The core of the device's acceptable performance is its substantial equivalence to a predicate device (Elecsys Vitamin D Assay K113546) and its traceability to a reference measurement procedure (ID-LC-MS/MS traceable to NIST SRM 2972).
Here's a table summarizing performance aspects:
| Performance Metric | Acceptance Criteria (Inferred/Implied) | Reported Device Performance (Elecsys Vitamin D total II) |
|---|---|---|
| Precision | Demonstrated acceptable repeatability and intermediate precision. | Precision study conducted per CLSI guideline EP5-A3. (Detailed results for mean, SD, and CV at various concentrations are provided in Table 1 under "Precision" for both repeatability and intermediate precision, showing performance comparable to or better than the predicate for the tested analyte levels and concentration ranges.) Example (Repeatability HS1): Mean 11.1 ng/mL, SD 0.725 ng/mL, CV 6.6%. Example (Intermediate HS1): Mean 11.1 ng/mL, SD 0.965 ng/mL, CV 8.7%. |
| Analytical Sensitivity | LoB, LoD, LoQ established and deemed acceptable. | LoB: 2 ng/mL (Same as predicate) |
| LoD: 3 ng/mL (Same as predicate) | ||
| LoQ: 5 ng/mL (Same as predicate) | ||
| Linearity/Reportable Range | Demonstrated linearity across the claimed measuring range. | Claimed measuring range: 5 - 100 ng/mL (Predicate: 5 - 60 ng/mL). Linearity study conducted according to CLSI guideline EP6-A using serum and plasma samples. Data analyzed for linear, quadratic, and cubic polynomials. (Specific linearity data not tabulated, but reported as successfully demonstrated.) |
| High Dose Hook Effect | No significant hook effect within relevant concentrations. | Assessed on cobas e 411 analyzer. "The hook concentration reported corresponds to the highest analyte concentration that generates a signal within the primary measuring range of the assay." (Implies no issue within the measuring range.) |
| Interferences | Limited interference from endogenous substances and common drugs. | HAMA: Assessed, recovery calculated. (Implied acceptable performance since no issues reported.) |
| Endogenous: Hemolysis (≤ 600 mg/dL), Bilirubin (≤ 66 mg/dL), Lipemia/Intralipid (≤ 300 mg/dL), Biotin (≤ 30 ng/mL), Serum albumin (≤ 7 g/dL), Cholesterol (≤ 300 mg/dL), Triglyceride (≤ 300 mg/dL), Rheumatoid Factor (< 1200 IU/mL), Total Protein (≤ 9 g/dL), IgG (≤ 7 g/dL) were tested. (Limits are specified, implying performance within these specified interference levels is acceptable). | ||
| Drugs: 16 commonly and 3 specially used pharmaceutical compounds tested. (Implied acceptable performance as no issues reported.) | ||
| Analytical Specificity/Cross-Reactivity | Acceptable cross-reactivity with other vitamin D metabolites. | 25-hydroxyvitamin D3: 100% (Same as predicate) |
| 25-hydroxyvitamin D2: 93.7% (Predicate: 92%) | ||
| 24, 25-dihydroxyvitamin D3: 13.7% (Predicate: 149%) - Note: Significant improvement from predicate, which showed high cross-reactivity with this metabolite. | ||
| 3-epi-25-hydroxyvitamin D3: 112.8% (Predicate: 91%) | ||
| 3-epi-25-hydroxyvitamin D2: 91.4% (Not specified for predicate) | ||
| Method Comparison to Reference Method | Strong correlation and low bias against the RMP. | Passing Bablok: y = 0.937x - 0.360, T = 0.902 (Candidate Assay against ID-LC-MS/MS RMP). (Implies good agreement with the reference method.) |
| Sample Matrix Comparison | Consistent results across different sample matrices (anticoagulants). | Compared serum, Li-Heparin, K2-EDTA-, K3-EDTA-plasma, and Plasma Gel Separation Tubes (Li-Heparin) using Passing/Bablok regression analysis. (Implied acceptable performance as no issues reported and data supports claim.) |
| Stability (Reagent, Calibrator, Control, Calibration) | Demonstrated stability for claimed storage conditions and intervals. | Reagent: After first opening (56 days at 2-8°C), on-board (28 days on analyzer). Real-time stability (shelf-life) ongoing. |
| Calibrator (CalSet): -20°C (12 weeks), 2-8°C (72 hours), on-board (5 hours), accelerated (35°C for 3 weeks), real-time (16 months). | ||
| Control (PreciControl): Reconstitution stability (-20°C for 32 days, 2-8°C for 73 hours, 20-25°C for 6 hours), accelerated (35°C for 3 weeks), real-time. | ||
| Calibration: Lot calibration stability (12 weeks/3 months), on-board calibration stability (7 days). | ||
| Reference Range | Establish a suitable reference interval for vitamin D sufficiency assessment. | Established reference interval: 7.61 - 55.5 ng/mL (for total population, based on 421 healthy adults). |
2. Sample Size Used for the Test Set and Data Provenance
The document describes various studies, each with its own sample size and provenance.
- Precision: 5 human serum samples (single donors, native and spiked).
- Analytical Sensitivity (LoB, LoD, LoQ):
- LoB: 1 blank sample (Vitamin D depleted human serum pool), 60 determinations.
- LoD: 5 low-level human serum sample pools (diluted), with replicates per run.
- LoQ: At least 4 low-level samples of serum, Li-heparin, K2-EDTA, K3-EDTA plasma, 25 replicates/sample/reagent lot.
- Linearity/Assay Reportable Range: 1 high analyte serum and K3-EDTA plasma sample (single donors, spiked), diluted to at least 9 concentrations.
- High Dose Hook Effect: 1 human serum sample spiked to approx. 10,000 ng/mL, dilution series performed.
- Human Anti-Mouse Antibodies (HAMA): HAMA-serum (HAMA L2) and related basis serum (without interferent).
- Endogenous Interferences: For each interfering substance, three human serum samples (single donors, native as well as spiked) containing low, mid, and high concentrations of 25-OH Vitamin D.
- Exogenous Interferences (Drugs): Two human serum samples (single donors, native as well as spiked) with analyte concentrations approx. 30 and 70 ng/mL.
- Analytical Specificity/Cross-Reactivity: Native human sera with approximate 25-hydroxyvitamin D concentrations of 25, 40, and 60 ng/mL (spiked with cross-reactants).
- Sample Matrix Comparison: Minimum of 40 serum/plasma pairs per sample material (single donors - native, diluted, spiked).
- Method Comparison to Predicate: 105 serum samples (single donors, native).
- Method Comparison to Reference Method: 111 native single donor patient serum samples.
- Reagent Stability: 5 human serum (HS) samples and 2 controls for run qualification (for each stability study). Human serum samples were single donors (native as well as spiked).
- Calibrator Stability: "on-test" and "reference materials" (specific number of human serum samples not detailed for each study, but implied to be sufficient for analytical validation).
- PreciControl Vitamin D total II (Value Assignment): Native human serum sample panel (single donors).
- Calibration Stability: 5 human serum (HS) samples and 2 controls (PreciControl Vitamin D total II). Pools of human serum samples (native as well as spiked).
- CalCheck Vitamin D total II (Value Assignment): Not explicitly stated, but performed for each lot.
- External (Clinical) Testing (Reference Range): Approximately 600 study subjects. After exclusions, 421 subjects were used for the reference range calculation.
Data Provenance:
- Clinical Samples for Reference Range: Prospectively collected from three US sites (northern, midwest, southern) during summer and winter seasons.
- Method Comparison to Reference Method: Patient serum samples from the Vitamin D Standardization and Certification Program (VDSCP) with assigned values by the RMP (Reference Measurement Procedure) at CDC.
- Analytical Studies (Precision, Linearity, Interference, etc.): Predominantly used human serum and plasma samples, often from "single donors" (native as well as spiked). The document implies these were collected appropriately for laboratory studies, likely from a variety of sources but not specifically detailed by country of origin beyond the US sites for the reference range study. All analytical studies appear to be retrospective in the sense that samples were acquired for the purpose of the study rather than being prospectively collected specifically for patient outcomes.
3. Number of Experts Used to Establish the Ground Truth for the Test Set and the Qualifications of Those Experts
The concept of "experts" as human readers establishing ground truth (common in imaging AI) is not directly applicable here. This device is an in vitro diagnostic (IVD) assay measuring a biomarker concentration.
- Ground Truth for Clinical Samples: For the Reference Range study (External/Clinical Testing), the "ground truth" is the measured Vitamin D total II concentration by the Elecsys Vitamin D total II assay itself, and the study then calculates the reference interval from this healthy population. The individuals themselves are not "experts" establishing a truth.
- Ground Truth for Analytical Studies:
- Traceability and Standardization: The assay is traced to the ID-LC-MS/MS 25-hydroxyvitamin D Reference Measurement Procedure (RMP) at the University of Ghent, which is further traceable to the National Institute of Standards and Technology (NIST) Standard Reference Material (SRM) 2972. This is the ultimate ground truth for Vitamin D concentration. These are established laboratory reference methods, not human expert consensus.
- Value Assignment of Controls and Calibrators: Values are assigned for PreciControl Vitamin D total II and CalCheck Vitamin D total II based on measurements with the Elecsys Vitamin D total II assay, which is itself traceable to the RMP. The "ground truth" for these materials is their assigned concentration based on the reference standard.
- Method Comparison: One comparison is against the ID-LC-MS/MS RMP at CDC, which serves as the independent reference standard (ground truth) for those 111 samples.
- Sample Pools/Spiking: For many analytical studies (e.g., precision, linearity, interference), test samples were prepared by pooling or spiking human serum/plasma. The "true" concentration for these samples is derived from the known amounts of spiked analyte or the consensus value of the pool, assuming the measurement method for verification is accurate and traceable.
In summary, the "ground truth" is established through highly controlled, traceable analytical reference methods and materials, not by human expert interpretation.
4. Adjudication Method for the Test Set
Adjudication methods (like 2+1, 3+1) are typically used in studies where multiple human readers interpret images or clinical data and discrepancies need to be resolved. This is not directly relevant to an IVD assay where the output is a quantitative concentration.
For the analytical studies, the "adjudication" is inherent in the rigorous statistical analysis and adherence to established CLSI guidelines (e.g., EP5-A3 for precision, EP17-A2 for LoB/LoD, EP6-A for linearity, EP07-A2 for interference). Any outlying data points would be addressed through statistical methods or re-testing according to study protocols, not through human adjudication of differing interpretations. The method comparison studies (against predicate and RMP) use statistical regression analyses (Deming, Passing Bablok) to assess agreement, rather than adjudication of individual results.
5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study Was Done
No, an MRMC comparative effectiveness study was not done. MRMC studies are specific to evaluating the impact of a medical device (often AI-powered) on the performance of human readers, commonly in imaging diagnostics. This device is an in vitro diagnostic (IVD) assay that directly measures a biomarker concentration. Its performance is evaluated analytically and against reference methods, not in how it assists human interpretation of complex cases.
Therefore, there is no effect size of how much human readers improve with AI vs. without AI assistance.
6. If a Standalone (i.e., Algorithm Only Without Human-in-the-Loop Performance) Was Done
Yes, the primary evaluation of the Elecsys Vitamin D total II assay is its standalone analytical performance. The entire "Non-Clinical Performance Evaluation" (Section 4) is dedicated to demonstrating the performance of the device itself (the assay on the cobas e 411 analyzer) across various metrics like precision, sensitivity, linearity, interference, specificity, and stability. The "Method Comparison to Reference Method" (4.11) particularly assesses its direct agreement with the gold standard RMP.
The "human-in-the-loop" component in this context is the laboratory technician operating the analyzer and interpreting the quantitative results within a clinical context (e.g., in aid of assessing vitamin D sufficiency). The performance evaluated here is solely that of the analytical system.
7. The Type of Ground Truth Used
The ground truth used for the device's validation is primarily external reference measurement procedures (RMPs) and reference materials.
- Primary Ground Truth: The ID-LC-MS/MS 25-hydroxyvitamin D Reference Measurement Procedure (RMP), specifically from the University of Ghent and CDC, traceable to the National Institute of Standards and Technology (NIST) Standard Reference Material (SRM) 2972. This is the metrological traceability chain that provides the "true" value for vitamin D concentration.
- Secondary Ground Truth/Comparators:
- Predicate Device: The Elecsys Vitamin D Assay (K113546) served as a comparator for demonstrating substantial equivalence.
- Healthy Adult Population (for reference range): For establishing the clinical reference range, the ground truth is simply the distribution of measured values from a carefully selected cohort of healthy individuals. This is a form of outcomes data in the sense of a normal physiological range.
- Known Spiked Concentrations: For many analytical studies (e.g., linearity, interference, specificity), samples were spiked with known amounts of analyte or interfering substances. The "true" concentration or the expected impact of the interferent serves as the ground truth for evaluating recovery or cross-reactivity.
8. The Sample Size for the Training Set
The document is a 510(k) summary for an IVD device, not an AI/ML device. Therefore, the concept of a "training set" for an algorithm in the machine learning sense is not applicable. The Elecsys Vitamin D total II assay is a chemical immunoassay, not a learning algorithm that undergoes a training phase with a dataset.
The parameters of the assay (reagent formulation, calibration curves) are established through developmental optimization and analytical validation, not through an iterative learning process with a "training set."
9. How the Ground Truth for the Training Set Was Established
As noted above, there is no "training set" in the machine learning sense. The assay's performance characteristics and calibration are established through the use of:
- Reference materials (calibrators): CalSet Vitamin D total II, which is traceable to the ID-LC-MS/MS RMP. These calibrators are used to establish the instrument-specific calibration curve for each reagent lot, ensuring the assay's accuracy against a known standard.
- Quality controls (controls): PreciControl Vitamin D total II, used to monitor the assay's performance and ensure it stays within predefined limits. Their assigned values are derived from measurements against the master calibration curve, which is ultimately traceable to the RMP.
Therefore, the "ground truth" during the development and routine use of the assay is established via the metrological traceability chain to international reference standards and reference measurement procedures.
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Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002
February 8, 2017
ROCHE DIAGNOSTICS LINDA MCCAMMACK REGULATORY PROGRAM MANAGER 9115 HAGUE ROAD INDIANAPOLIS IN 46250
Re: K162840
Trade/Device Name: Elecsys Vitamin D total II. CalSet Vitamin D total II. CalCheck Vitamin D total II, PreciControl Vitamin D total II Regulation Number: 21 CFR 862.1825 Regulation Name: Vitamin D test system Regulatory Class: II Product Code: MRG, JIT, JJX Dated: December 22, 2016 Received: December 23, 2016
Dear Linda McCammack:
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.
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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,
Katherine Serrano -S
For : Courtney H. Lias, Ph.D. Director Division of Chemistry and Toxicology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health
Enclosure
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Indications for Use
510(k) Number (if known) K162840
Device Name Elecsys Vitamin D total II CalCheckVitamin D total II CalSet Vitamin D total II PreciControl Vitamin D total II
Indications for Use (Describe)
Elecsys Vitamin D total II
The Elecsys Vitamin D total II assay is intended for the quantitative determination of total 25-hydroxyvitamin D in human serum and plasma. This assay is to be used as an aid in the assessment of vitamin D sufficiency in adults. The electrochemiluminescence binding assay is intended for use on the cobas e 411 immunoassay analyzer.
CalSet Vitamin D total II
CalSet Vitamin D total II is used for calibrating the quantitative Elecsys Vitamin D total II assay on the cobas e 411 immunoassay analyzer.
PreciControl Vitamin D total II
PreciControl Vitamin D total II is used for quality control of the Elecsys Vitamin D total II assay on the cobas e 411 immunoassay analyzer.
CalCheck Vitamin D total II
This CalCheck set is an assayed control for use in calibration verification of the assay range established by the Elecsys Vitamin D total II reagent on the cobas e 411 immunoassay analyzer.
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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Elecsys Vitamin D total II 510(k) Summary
This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of 21 CFR 807.92.
In accordance with 21 CFR 807.87, Roche Diagnostics hereby submits official notification as required by Section 510(k) of the Federal Food, Drug and Cosmetics Act of our intention to market the device described in this Premarket Notification 510(k).
The purpose of this Traditional 510(k) Premarket Notification is to obtain FDA review and clearance for the Elecsys Vitamin D total II Test System.
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| Submitter Name | Roche Diagnostics |
|---|---|
| Address | 9115 Hague RoadP.O. Box 50416Indianapolis, IN 46250-0457 |
| Contact | Linda McCammackPhone: (317) 521-7144FAX: (317) 521-2324Email: linda.mccammack@roche.com |
| Date Prepared | September 19, 2016 |
| Proprietary Name | 1- Elecsys Vitamin D total II2-CalSet Vitamin D total II3- PreciControl Vitamin D total II4- CalCheck Vitamin D |
| Common Name | 1-Vitamin D total2 -CalSet Vitamin D total II3 - PreciControl Vitamin D total II4 - CalCheck Vitamin D |
| Classification Name | 1 - System, Test, Vitamin D,2 - Calibrator, Secondary,3 - Single (Specified) Analyte Controls (Assayed and Unassayed),4 - Single (Specified) Analyte Controls (Assayed and Unassayed) |
| Product Codes,Regulation Numbers | 1 -MRG, 862.18252- JIT, 862.11503- JJX, 862.16604- JJX, 862.1660 |
| Predicate Devices | Elecsys Vitamin D Assay (K113546) |
| Establishment Registration | For the Elecsys Vitamin D total II, the establishment registration numbers:Roche Diagnostics GmbH in Mannheim, Germany is 9610126,Roche Diagnostics Penzberg, Germany is 9610529Roche Diagnostics in the United States is 1823260 |
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DEVICE DESCRIPTION 1.
Elecsys Vitamin D total II is a second generation assay by Roche Diagnostics for the in vitro quantitative determination of 25-hydroxyvitamin D in human serum and plasma. It is intended for use on the cobas e 411 immunoassay analyzer. The cobas e family of analyzers employs the electrochemiluminescence "ECLIA" technology. The assay is a 27 minute assay utilizing a competition principle and a pretreatment step to release the bound 25-hydroxyvitamin D from the vitamin D binding protein.
Results are determined via a calibration curve which is instrument specifically generated by 2-point calibration against the master curve for that reagent lot.
Reagents 1.1.
The Elecsys Vitamin D total II reagent working solutions include:
- Pretreatment rackpack (kit placed on instrument) .
- Pretreatment Reagent 1 (Dithiothreitol) -
- Pretreatment Reagent 2 (Sodium hydroxide) -
- Rackpack (kit placed on instrument) .
- -Streptavidin-coated microparticles
- -Reagent 1 (ruthenium labeled vitamin D binding protein)
- Reagent 2 (biotinylated 25-hydroxyvitamin D) -
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1.2. Calibrator
The assay will be calibrated using the CalSet Vitamin D total II, which is a two concentration level set of lyophilized human serum matrix that are traceable to the ID-LC-MS/MS 25-hydroxyvitamin D Reference Measurement Procedure. The ID-LC-MS/MS is traceable to the National Institute of Standards and Technology Standard Reference Material 2972. The CalSet includes:
- Cal 1 (approximately 2 ng/mL 25-hydroxyvitamin D in human serum matrix) .
- Cal 2 (approximately 45 ng/mL 25-hydroxyvitamin D in human serum matrix) .
1.3. Control
PreciControl Vitamin D total II is used for the quality control verification of the performance of the assay on the cobas e 411 analyzer. The control materials are lyophilized serum based on human serum in two concentration ranges.
1.4. Calibration Verification Materials
CalCheck Vitamin D total II is used for the calibration verification and the assessment of the measuring range as needed by the laboratory certification agencies such as College of American Pathologists or CLIA certification. The CalChecks are a customer convenience product and not required to assess the performance.
INTENDED USE 2.
Following are the Intended Uses for the Elecsys Vitamin D total II Test System.
Elecsys Vitamin D total II 2.1.
The Elecsys Vitamin D total II assay is intended for the quantitative determination of total 25-hydroxyvitamin D in human serum and plasma. This assay is to be used as an aid in the assessment of vitamin D sufficiency in adults. The electrochemiluminescence binding assay is intended for use on the cobas e 411 immunoassay analyzer.
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CalSet Vitamin D total II 2.2.
CalSet Vitamin D total II is used for calibrating the quantitative Elecsys Vitamin D total II assay on the cobas e 411 immunoassay analyzer.
PreciControl Vitamin D total II 2.3.
PreciControl Vitamin D total II is used for quality control of the Elecsys Vitamin D total II assay on the cobas e 411 immunoassay analyzer.
2.4. CalCheck Vitamin D total II
The CalCheck set is an assayed control for use in calibration verification and for use in the verification of the assay range established by the Elecsys Vitamin D total II reagent on the cobas e 411 immunoassay analyzer.
TECHNOLOGICAL CHARACTERISTICS 3.
Elecsys Vitamin D total II utilizes electrochemiluminescence "ECLIA" technology for the quantitative determination of 25-hydroxyvitamin D for the assessment of vitamin D sufficiency on the cobas e 411 immunoassay analyzer.
The following tables compare the Elecsys Vitamin D total II with its predicate device, Elecsys Vitamin D Assay (K113546).
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Assay Comparison Table 1:
| Feature | Predicate Device:Elecsys Vitamin D Assay (K113546) | Candidate Device:Elecsys Vitamin D total II |
|---|---|---|
| Intended Use/Indications for Use | This Vitamin D assay in intended for thequantitative determination of total25-hydroxyvitamin D in human serum andplasma. This assay is to be used as an aid inthe assessment of vitamin D sufficiency inadults. The electrochemiluminescencebinding assay is intended for use on Elecsysand cobas e immunoassay analyzers. | This assay is intended for the quantitativedetermination of total 25-hydroxyvitaminD in human serum and plasma. This assayis to be used as an aid in the assessment ofvitamin D sufficiency in adults. Theelectrochemiluminescence binding assay isintended for use on the cobas e 411immunoassay analyzer. |
| Assay Method | Competition principle binding protein | Same |
| Detection Method | Electrochemiluminescence | Same |
| Applications/TestTime | 27 minutes | Same |
| Instrument Platform | Elecsys 2010, MODULAR ANALYTICSE170, cobas e 411, cobas e 601, andcobas e 602 | cobas e 411 |
| Sample/Reagent Ratio | 15 µL / 185 µL | 20 µL / 180 µL |
| Sample Type/Matrix | Human serum, plasma | Same |
| Sample Anticoagulants | Li-heparin, K2-EDTA, K3-EDTA,Li-heparin PST. | Li-heparin, K2-EDTA,K3-EDTA, PST tubes for correspondingplasma types. |
| Calibrator | Vitamin D CalSet | CalSet Vitamin D total II |
| Calibration Method | 2-point calibration based on master curve forspecific reagent lot | Same |
| Calibration Interval | Calibration must be performed once perreagent lot using fresh reagent (< 24hours since registered). Renewedcalibration:• After 1 month (28 days) using thesame reagent lot• After 7 days (when using the samereagent kit on the analyzer)• As required e.g. quality controlfindings outside the defined limits | Calibration must be performed once perreagent lot using fresh reagent (< 24hours since registered). Renewedcalibration:• After 3 months (12 weeks) usingthe same reagent lot• After 7 days (when using the samereagent kit on the analyzer)• As required e.g. quality controlfindings outside the defined limits |
| Controls | PreciControl Varia | PreciControl Vitamin D total II |
| Traceability/Standardization | LC-MS/MS traceable to NIST SRM 2972 | ID-LC-MS/MS RMP traceable toNIST 2972. |
| Reagent Stability | • Unopened at 2-8°C up to the statedexpiration date• After opening at 2-8°C 56 days (8weeks)• On the analyzer 21 days | • Unopened at 2-8°C up to thestated expiration date• After opening at 2-8°C 56 days (8weeks)• On the analyzer 28 days (4 weeks) |
| Measuring Range | 5 - 60 ng/mL | 5 - 100 ng/mL |
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| Feature | Predicate Device:Elecsys Vitamin D Assay (K113546)Elecsys 2010 and cobas e 411 analyzers: | Candidate Device:Elecsys Vitamin D total II | ||||||
|---|---|---|---|---|---|---|---|---|
| Repeatability | Repeatability | |||||||
| mean[ng/mL] | SD[ng/mL] | CV[%] | mean[ng/mL] | SD[ng/mL] | CV[%] | |||
| Precision | HS1 | 6.20 | 0.445 | 7.2 | HS1 | 11.1 | 0.725 | 6.6 |
| HS2 | 11.2 | 1.42 | 5.1 | HS2 | 20.8 | 0.849 | 4.1 | |
| HS3 | 23.2 | 0.666 | 2.9 | HS3 | 25.6 | 0.774 | 3.0 | |
| HS4 | 43.5 | 1.04 | 2.4 | HS4 | 47.5 | 0.749 | 1.6 | |
| HS5 | 52.6 | 0.858 | 1.6 | HS5 | 92.6 | 1.76 | 1.9 | |
| Intermediate | Intermediate | |||||||
| HS1 | 6.20 | 0.640 | 10.3 | HS1 | 11.1 | 0.965 | 8.7 | |
| HS2 | 11.2 | 0.849 | 7.6 | HS2 | 20.8 | 1.09 | 5.2 | |
| HS3 | 23.2 | 1.24 | 5.3 | HS3 | 25.6 | 1.43 | 5.6 | |
| HS4 | 43.5 | 1.46 | 3.4 | HS4 | 47.5 | 1.77 | 3.7 | |
| HS5 | 52.6 | 1.50 | 2.9 | HS5 | 92.6 | 2.40 | 2.6 | |
| LoB | 2 ng/mL | Same | ||||||
| LoD | 3 ng/mL | Same | ||||||
| LoQ | 5 ng/mL | Same | ||||||
| 25-hydroxyvitamin D3 | 100% | 25-hydroxyvitamin D3 | 100% | |||||
| 25-hydroxyvitamin D2 | 92% | 25-hydroxyvitamin D2 | 93.7% | |||||
| Analytical Specificity | 24, 25-dihydroxyvitamin D3 | 149% | 24, 25-dihydroxyvitamin D3 | 13.7% | ||||
| C3-epimer of 25-hydroxyvitamin D3 | 91% | 3-epi-25-hydroxyvitamin D3 | 112.8% | |||||
| 3-epi-25-hydroxyvitamin D2 | 91.4% | |||||||
| Method ComparisonCandidate Assayagainst ID-LC-MS/MSRMP | Deming Regression$y = 0.954x - 0.707$$r = 0.982$Using 111 samples with sample concentrations between approximately 5.6 ng/mL and 93 ng/mL | Passing Bablok$y = 0.937x - 0.360$$T = 0.902$ | ||||||
| Method ComparisonCandidate Assayagainst PredicateDeviceElecsys Vitamin DAssay | Deming Regression$y = 0.849x + 0.983$$r = 0.955$Using 105 samples with sample concentrations between approximately 6.1 ng/mL and 56 ng/mL | Passing Bablok$y = 0.867x + 0.724$$T = 0.840$ | ||||||
| Limitations | Hemolysis | ≤ 2 g/L | Hemolysis | ≤ 600 mg/dL | ||||
| Bilirubin | ≤ 66 mg/dL | Bilirubin | ≤ 66 mg/dL | |||||
| Lipemia (Intralipid) | ≤ 400 mg/dL | Intralipid | ≤ 300 mg/dL | |||||
| Biotin | ≤ 70 ng/mL | Biotin | ≤ 30 ng/mL |
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| Feature | Predicate Device:Elecsys Vitamin D Assay (K113546) | Candidate Device:Elecsys Vitamin D total II | ||
|---|---|---|---|---|
| Serum albumin | $\leq$ 7 g/dL | Serum albumin | $\leq$ 7 g/dL | |
| Cholesterol | $\leq$ 380 mg/dL | Cholesterol | $\leq$ 300mg/dL | |
| Triglyceride | $\leq$ 300 mg/dL | |||
| Rheumatoid Factor | $<$ 1200 IU/mL | |||
| Total Protein | $\leq$ 9 g/dL | |||
| IgG | $\leq$ 7 g/dL |
Table 2: Calibrator Comparison
| Feature | Predicate Device | Candidate Device |
|---|---|---|
| Intended Use/Indications for Use | Vitamin D CalSet is used for calibrating thequantitative Elecsys Vitamin D assay on theElecsys and cobas e immunoassay analyzers. | CalSet Vitamin D total II is used forcalibrating the quantitative ElecsysVitamin D total II on the cobas e 411immunoassay analyzer. |
| Analyte | 25-hydroxyvitamin D | 25-hydroxyvitamin D3 |
| Matrix | Human serum matrix with added25-hydroxyvitamin D | Human serum matrix with added25-hydroxyvitamin D3 |
| Levels | Two | Same |
| Target Ranges | Cal 1: approximately 2 ng/mLCal 2: approximately 37 ng/mL | Cal 1: approximately 2 ng/mLCal 2: approximately 45 ng/mL |
| Format | Lyophilized | Same |
| Handling | Carefully dissolve the contents of one bottle byadding exactly 1.0 mL of distilled or deionizedwater and allow to stand closed for 15 minutes toreconstitute. Mix carefully, avoiding the foamformation. | Same |
| Storage and Stability | Reconstituted calibrators:• At 2-8°C – 120 hours• At -20°C – 90 days (freeze only once)• On the cobas e 411 analyzer – up to 5 hours | Reconstituted calibrators:• At 2-8°C – 72 hours• At -20°C – 12 weeks (freeze only once)• On the cobas e 411 analyzer – up to 5 hours |
Table 3: Control Comparison
| Feature | Predicate Device | Candidate Device |
|---|---|---|
| Intended Use/Indications for Use | PreciControl Varia is used for the quality controlof Elecsys immunoassays on the Elecsys andcobas e immunoassay analyzers. | PreciControl Vitamin D total II is usedfor quality control of the ElecsysVitamin D total II assay on thecobas e 411 immunoassay analyzer. |
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| Feature | Predicate Device | Candidate Device |
|---|---|---|
| Analyte | Vitamin B12FerritinFolateβ-CTxOsteocalcinParathyroid hormone25-hydroxyvitamin DCalcitonin | 25-hydroxyvitamin D |
| Matrix | Human serum matrix | Same |
| Levels | Three | Two |
| Target Ranges | Target range for the 25-hydroxyvitamin D:• Level 0: Approximately 12.8 ng/mL• Level 1: Approximately 17 ng/mL• Level 2: Approximately 32 ng/mL | Target range:• Level 1: Approximately 13 ng/mL• Level 2: Approximately 30 ng/mL |
| Format | Lyophilized | Same |
| Handling | Carefully dissolve the contents of one bottle byadding exactly 3.0 mL of distilled or deionizedwater and allow to stand closed for 30 minutes toreconstitute. Mix carefully, avoiding the foamformation. | Carefully dissolve the contents of onebottle by adding exactly 1.0 mL ofdistilled or deionized water and allow tostand closed for 15 minutes toreconstitute. Mix carefully, avoiding thefoam formation. |
| Storage and Stability | Reconstituted control serum:• At -20°C – 31 days (freeze only once)• At 2-8°C – 72 hours• At 20-25°C - up to 5 hours | Same |
Table 4: CalCheck Comparison
| Feature | Predicate Device | Candidate Device |
|---|---|---|
| IntendedUse/Indications forUse | The Elecsys Vitamin D CalCheck 5 is anassayed control for use in calibration verificationand for use in the verification of the assay rangeestablished by the Elecsys Vitamin D reagent onthe Elecsys and cobas e immunoassay analyzers. | CalCheck Vitamin D total II is anassayed control for use in calibrationverification and for use in theverification of the assay rangeestablished by the Elecsys Vitamin Dtotal II reagent on the cobas e 411immunoassay analyzer. |
| Analyte | 25-hyroxyvitamin D3 | Same |
| Matrix | Human serum | Same |
| Levels | Five | Same |
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| Feature | Predicate Device | Candidate Device |
|---|---|---|
| Target Ranges | Approximate target concentrations:• ≤ 5.00 ng/mL• 10.5 - 19.5 ng/mL• 21.0 - 39.0 ng/mL• 33.6 - > 60.0 ng/mL• 42.0 - > 60.0 ng/mL | Approximate target concentrations:• ≤ 5 ng/mL• 17.5 - 22.4 ng/mL• 46.5 - 54.4 ng/mL• 75.5 - 84.4 ng/mL• 94.5 - > 100 ng/mL |
| Format | Lyophilized | Same |
| Handling | Reconstitute the contents of Check 1, Check 2,Check 3, Check 4 and Check 5 with exactly 1.0mL distilled or deionized water. Allow thebottles to stand closed for 15 minutes. Mixgently by inversion to ensure homogeneity. | Same |
| Storage and Stability | Reconstituted control serum:• At 20-25°C – 5 hours | Same |
NON-CLINICAL PERFORMANCE EVALUATION 4.
Non-clinical performance evaluation for Elecsys Vitamin D total II executed with the study briefly summarized.
4.1. Precision
Repeatability and Intermediate Precision 4.1.1.
Precision was evaluated on a single cobas e 411 analyzer according to CLSI guideline EP5-A3 using one reagent lot for evaluation. The protocol consisted of testing 2 replicates of each of two levels of control, PreciControl Vitamin D total II and five human sera per run, 2 runs per day for 21 days. The samples were run in randomized order on the cobas e 411 analyzer. Serum samples used were all single donors (native as well as spiked).
Analytical Sensitivity 4.2.
Limit of Blank (LoB) 4.2.1.
Experimental Design included three reagent lots evaluated on one cobas e 411 analyzer, six runs over three or more days with one blank sample with ten replicates per run. The zero-level (blank) sample used was a Vitamin D depleted human serum sample pool. In total, 60 determinations for analyte free samples have been obtained. The LoB was calculated according to CLSI EP17-A2.
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4.2.2. Limit of Detection (LoD)
Experimental Design included three reagent lots evaluated on one cobas e 411 analyzer, six runs over three or more days with five samples with two replicates/sample/run. The five samples were low-level human serum sample pools (diluted). A pooled estimate of the precision (SD total) for the 5 low-level samples was calculated. The LoD was calculated according to CLSI EP17-A2.
Limit of Quantitation (LoQ) 4.2.3.
Experimental Design included three reagent lots evaluated on one cobas e 411 analyzer, one run per day over five days. Five replicates per each sample per run with at least four low level samples of serum, Li -heparin, K2-EDTA and K3-EDTA plasma and 25 replicates/sample/reagent lot.
The mean value and the intermediate precision (CV) and standard deviation (SD) for each LoQ sample were calculated.
LoQ samples were sorted according to the concentration of their measured mean value. LoQ is defined as the mean value of that sample which is the first that fulfills the specification for the intermediate precision and for which no sample with higher concentration exists that exceeds this specification.
Linearity/Assay Reportable Range 4.3.
The linearity study was conducted on the Elecsys Vitamin D total II assay to demonstrate that measurements across the claimed measuring range for each parameter are linear. The study was performed according to CLSI guideline EP6-A using serum and plasma samples on the cobas e 411 immunoassay analyzer.
A high analyte serum and K3-EDTA plasma sample (single donors, spiked) was diluted with a depleted low analyte human serum. At least 9 concentrations (dilutions) throughout the measuring range were prepared. Samples were assayed in 3-fold determination within a single run.
The linearity data were analyzed with regards to linear, quadratic and cubic polynomials according to CLSI EP6-A.
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High Dose Hook Effect 4.4.
The high-dose hook effect of the Elecsys Vitamin D total II assay was assessed on the cobas e 411 analyzer in two-fold determination. One human serum sample was spiked with analyte to achieve a high 25-OH Vitamin D concentration of approx. 10,000 ng/mL. A dilution series was performed using a low level analyte (Vitamin D depleted) serum. The hook concentration reported corresponds to the highest analyte concentration that generates a signal within the primary measuring range of the assay.
Human Anti-Mouse Antibodies (HAMA) 4.5.
The effect of the presence of human anti-mouse antibodies on the Elecsys Vitamin D total II assay was assessed on the cobas e 411 immunoassay analyzer.
The specified HAMA-serum (HAMA L2) and the related basis serum (without interferent) was measured in two-fold determination each. Recovery of the HAMA-serum compared to the basis serum was calculated.
Endogenous Interferences 4.6.
Hemolysis/Bilirubin/Lipemia/Biotin 4.6.1.
The purpose of this study was to evaluate endogenous substances for potential interference with the parameters measured with the Elecsys Vitamin D total II on the cobas e 411 immunoassay analyzer using human serum samples (single donors, native as well as spiked).
For each interfering substance three human serum samples containing low, mid and high concentrations of 25-OH Vitamin D were tested in accordance with CLSI EP07-A2.
Triglycerides Interference 4.6.2.
One aliguot of each serum sample was spiked with the interfering substance (Triglyceride concentrate), another aliquot was spiked with the same volume of isotonic NaCl solution (dilution pool). The interfering pool was then diluted into the dilution pool in 10% increments. The recovery for each sample was calculated by comparison to the reference sample.
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Rheumatoid Factors Interference 4.6.3.
One aliquot of each serum sample was spiked with the interfering substance, and another aliquot with the same volume of the solvent (buffer matrix) of the interfering substance (dilution pool). The interfering pool was then diluted into the dilution pool in 10% increments. The recovery for each sample was calculated by comparison to the reference sample.
Total Protein/Albumin/Immunoglobulin (IgG)/Cholesterol Interference 4.6.4.
One aliquot of each serum sample was spiked with the interfering substances. Another aliquot without any additives (since the interfering substance is a lyophilisate) is used as dilution pool. The interfering pool was then diluted into the dilution pool in 10% increments.
Analytical Specificity/Cross-Reactivity 4.7.
A cross-reactivity study was conducted according to CLSI-EP7-A2 to evaluate the potential cross-reactivity of the assay with other vitamin D metabolites that occur in the human body during anabolism and catabolism. The potential cross-reactants were added at defined concentrations to native human sera with approximate 25-hydroxyvitamin D concentrations of 25, 40 and 60 ng/mL and analyzed with Elecsys Vitamin D total II on the cobas e 411analyzer. The concentrations of the cross-reactants where chosen i) to ensure results within the measuring range depending on their expected cross-reactivity and ii) to reflect at least a >10 times higher concentration as typically found in native serum. Results from these spiked serum samples were matched against the unspiked references and the % cross-reactivity (normalized and nonnormalized) was calculated.
The analysis was done in nmol/L to take into account i) the different molecular weights of the vitamin D metabolites and ii) the 1:1 equimolar binding of metabolite and vitamin D binding protein.
4.8. Exogenous Interferences – Drugs
The effect on quantitation of analyte in the presence of drugs was determined by comparing values obtained from samples spiked with 16 commonly and 3 specially used pharmaceutical compounds with the reference sample (unspiked). Two human serum samples (single donors,
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native as well as spiked) were used and tested on the cobas e 411 immunoassay analyzer. The analyte concentration of the samples were approximately 30 and 70 ng/mL.
The drug concentrations tested are according to the recommendation (if available) given in the CLSI guideline EP7-A2. When concentrations are not given in the guideline, concentrations of at least 3-times of the maximum recommended daily dose were tested.
The two serum samples were divided into aliquots and spiked with the potential interferents.
The reference sample without interferent was spiked with the respective amount of solvent only.
The Elecsys Vitamin D total II concentration of the spiked aliquots was determined in 10-fold determination and compared to the Vitamin D total II result determined for the reference aliquot (also in 10-fold determination) on one cobas e 411 immunoassay analyzer.
Sample Matrix Comparison 4.9.
The effect on quantitation of analyte in the presence of anticoagulants with the
Elecsys Vitamin D total II Immunoassay was determined by comparing values obtained from samples (single donors - native, diluted as well as spiked) drawn into serum, Li-Heparin, K2-EDTA- , K3-EDTA-plasma primary tubes and Plasma Gel Separation Tubes (Li-Heparin).
A minimum of 40 serum/plasma pairs per sample material were tested in singleton with one reagent lot on one cobas e 411 immunoassay analyzer. Potential effects are assessed by Passing/Bablok regression analysis.
4.10. Method Comparison to Predicate
A method comparison was performed using the Elecsys Vitamin D total II assay (candidate device, Y) and the Elecsys Vitamin D Assay (predicate device, X) to assess the bias between the two assays (for information only).
A total of 105 serum samples (single donors, native) were measured in singleton on the cobas e 411 analyzer in one run covering the entire measuring range. Vitamin D values ranged between 6.1 and 55.9 ng/mL for the Reference Method (X) according to the recommendations of CLSI EP9-A2.
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4.11. Method Comparison to Reference Method
A total of 111 native single donor patient serum samples (reference sample set provided by the Vitamin D Standardization and Certification Program with assigned values by the RMP at CDC, independent from the samples used for standardization) were measured in singleton on the cobas e 411 analyzer in one run covering the entire measuring range. 25-hydroxyvitamin D values ranged between 5.64 and 92.8 ng/mL for the Reference Method.
4.12. Reagent Stability
To test reagent stability, three studies were executed with two studies completed including:
- Study 1: Reagent stability after first opening at 2-8°C (56 days) .
- Study 2: On board reagent stability (28 days) .
- Study 3: A real-time stability study is ongoing to support shelf-life stability claim. .
4.12.1. Reagent Stability After First Opening
Reagent stability after first opening for the Elecsys Vitamin D total II assay was tested on one cobas e 411 immunoassay analyzer. A fresh reagent rackpack was placed on the analyzer and calibrated. Reference values for the samples tested were determined. After initial measurement the kit was removed from the analyzer and kept at 2-8 °C up to 64 days. After 64 days the kit was placed on the analyzer again, calibrated and the test samples were determined. Samples tested in duplicate include five human serum (HS) samples and two controls (PreciControl Vitamin D total II) for run qualification. The human serum samples used were all single donors (native as well as spiked).
4.12.2. On-board Reagent Stability
Reagent On-board Stability for the Elecsys Vitamin D total II assay was tested on one cobas e 411 immunoassay analyzer. A fresh Reagent Rack-Pack was placed on the analyzer and calibrated. All samples were measured on day 0. On day 8, 15, 22, 29 and day 36, the same samples were measured with the same reagent kit kept at 20℃ ± 3℃ (on-board condition) using the calibration curves established on day 0, 8, 15, 22 and 29, respectively. Samples tested include
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five human serum (HS) samples and two controls (PreciControl Vitamin D total II) for run qualification. Each sample was tested in two-fold determination. The human serum samples used were all single donors (native as well as spiked).
4.12.3. Real-time Stability
In the ongoing real-time stability study, the Elecsys Vitamin D total II reagent is stored at 2-8°C. The stored assay reagents are tested at time point T=0 (after manufacturing) and at specified intervals over the shelf life of the device up to the planned shelf life plus one month. Testing will be performed using three human serum samples (stored at -80°C). The average on-test recovery value will be calculated as percent recovery compared to the reference value measured at T=0 for the human serum samples.
4.13. Calibrator Stability
Five studies were performed in order to verify the stability claims for the CalSet Vitamin D total II.
- Study 1: Stability at -20℃ after reconstitution .
- Study 2: Stability at 2-8°C after reconstitution .
- Study 3: On-board stability at 20-25℃ after reconstitution .
- . Study 4: Accelerated stability at 35°C
- Study 4: Real-time stability .
4.13.1. Stability at -20°C after reconstitution
The on-test and reference materials were tested in duplicate on the cobas e 411 immunoassay analyzer. The on-test material was reconstituted and stored in closed vials for 14 weeks at -20°C. The on-test signal recovery was calculated as percent of the reference value.
4.13.2. Stability at 2-8°C after reconstitution
The on-test and reference materials were tested in duplicate on the cobas e 411 immunoassay analyzer. The on-test material was reconstituted and stored in closed vials for 73 hours at 2-8°C. The on-test signal recovery was calculated as percent of the reference value.
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4.13.3. On-board stability at 20-25°C after reconstitution
The on-test and reference materials were tested in duplicate on the cobas e 411 immunoassay analyzer. The on-test material was reconstituted and stored in open vials for 7 hours at 20-25°C. The on-test signal recovery was calculated as percent of the reference value.
4.13.4. Accelerated stability at 35°C
The on-test and reference materials were tested in duplicate on the cobas e 411 immunoassay analyzer. The on-test material was stored lyophilized (as supplied to the user) at 35°C for 3 weeks. The reference material was a set of CalSet Vitamin D total II (stored at 2 - 8°C). After 3 weeks, the on-test and reference materials were tested in duplicate. The on-test signal recovery was calculated as a percent of the reference value.
4.13.5. Real-time stability
In the on-going real-time stability study, the CalSet Vitamin D total II test material is stored at 2-8°C. The CalSets are tested in duplicate at specified intervals over the shelf life of the device up to the planned shelf life plus one month (16 months).
Real-time stability is calculated based on the recovery of signal of stressed calibrator (stored at 2-8°C) vs. unstressed calibrator (stored at -80°C). At the specified intervals over the shelf life, the mean value of the stressed calibrator was calculated as percent recovery of the unstressed value (each tested in duplicates at the same time point).
4.14. PreciControl Vitamin D total II
4.14.1. Value Assignment
The PreciControl Vitamin D total II assigned values are determined with the Elecsys Vitamin D total II assay. A native human serum sample panel (single donors) with RMP-assigned target values (RMP = Reference Measurement Procedure: ID-LC-MS/MS of the University of Ghent) is used for Reference Standardization to assign values to the Master Calibrators (sample curve consisting of human serum samples from single donors covering the entire measuring range). These Master Calibrators are for routine use in subsequent standardizations. The assigned values for PreciControl Vitamin D total II are read from the master calibration curve.
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Values are assigned for each lot of PreciControl Vitamin D total II in combination with each assay reagent lot available. The controls are run in duplicate on at least two (2) modules of the master analyzer platform. The assigned value of each control level is defined as the median value obtained over at least six (6) determinations of the respective control level.
4.14.2. Stability
Three studies were performed in order to verify the stability claims for the PreciControl Vitamin D total II.
- Study 1: Stability after Reconstitution .
- Study 2: Accelerated Stability .
- Study 3: Real-time Stability .
4.14.2.1. Stability after reconstitution
The on-test and reference materials were tested in duplicate. The on-test material was reconstituted and stored for 32 days at -20℃, 73 hours at 2 - 8℃ and for 6 hours at 20 - 25℃. The reference material was freshly reconstituted PreciControl Vitamin D total II stored at 2 – 8°C. The deviation of the on-test material compared to the reference value was calculated. The PreciControl Vitamin D total II was evaluated in duplicate on the cobas e 411 immunoassay analyzer.
4.14.2.2. Accelerated Stability
The on-test material was stored lyophilized (as supplied to the user) at 35°C for 3 weeks. The reference material was PreciControl Vitamin D total II stored at 2 - 8℃. After 3 weeks, the ontest and reference material was tested in duplicate. The deviation of the on-test material compared to the reference value was calculated. The PreciControl Vitamin D total II was evaluated in duplicate on the cobas e 411 immunoassay analyzer.
4.14.2.3. Real-time Stability
In the on-going real-time stability study, the PreciControl Vitamin D total II on-test material is stored at 2 - 8°C. The controls are tested at specified intervals over the shelf life of the device up
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to the planned shelf life plus one month. Data for the time-points after manufacturing, 7, 10, 13 and 16 months tested in duplicate will be available. The deviation of the on-test material compared to the reference value (stored at -80°C) will be calculated.
4.15. Calibration Stability
To test calibration stability, two studies were completed, including:
- . Study 1: Lot calibration stability
- Study 2: On-board calibration stability .
- 4.15.1. Lot Calibration Stability
Calibration of an Elecsys Vitamin D total II reagent lot is recommended every 12 weeks (3 months). During that time period fresh reagent kits of the same lot can be used without calibration using the calibration curve of the day 0 reagent kit. Elecsys Vitamin D total II was calibrated with a fresh reagent kit on day 0 using a cobas e 411 immunoassay analyzer. After 13 weeks a new reagent kit of the same lot was used and recovery of samples was determined using the calibration curve of day 0. Five human serum (HS) samples and two controls (PreciControl Vitamin D total II) for run qualification were tested; each sample was tested with two-fold determination. Pools of human serum samples were used (native as well as spiked).
4.15.2. On-board Calibration Stability
Elecsys Vitamin D total II reagent kits can be stored on board of the analyzers for up to 7 days without a new calibration. Reagent On-board Calibration stability for the Elecsys Vitamin D total II assay was tested on one cobas e 411 immunoassay analyzer. A fresh Reagent Rack-Pack was placed on the analyzer and calibrated. All samples were measured on day 0. On day 8 the same samples were measured with the same reagent kit kept at 20°C ± 3°C (on-board condition) using the calibration curve established on day 0. Samples tested include five human serum (HS) samples and two controls (PreciControl Vitamin D total II) for run qualification. Each sample was tested in two-fold determination. Pools of human serum samples were used (native as well as spiked).
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4.16. CalCheck Vitamin D total II
The CalCheck Vitamin D total II matrix, human serum, is identical to the Master Calibrators (human serum sample panel) used for the Elecsys Vitamin D total II assay.
4.16.1. Value Assignment
For each CalCheck Vitamin D total II lot manufactured, the CalChecks are run in duplicate on at least two (2) modules with at least two runs. The assigned value of each CalCheck is defined as the median value obtained over at least six (6) determinations (runs) of the respective CalCheck.
4.16.2. Stability
Two studies were performed in order to verify the stability claims for the CalCheck Vitamin D total II.
- Study 1: Open Vial Stability .
- Study 2: Accelerated Stability .
- Study 3: Real-time Stability .
4.16.2.1. Open Vial Stability
The on-test and reference materials were tested in duplicate. The on-test material was reconstituted and stored for 6 hours at 25℃ (in an open vial). The reference material was a freshly reconstituted set of CalChecks. The on-test recovery was calculated as a percent of the reference value.
4.16.2.2. Accelerated Stability
The on-test material was stored lyophilized (as supplied to the user) at 35°C for 3 weeks. The reference material was a set of CalChecks (stored at 2-8°C). After 3 weeks, the test and reference materials were tested in duplicate. The on-test recovery was calculated as a percent of the reference value. One CalCheck Vitamin D total II lot was evaluated in duplicate on the cobas e 411 immunoassay analyzer.
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4.16.2.3. Real-time Stability
In the on-going real-time stability study, the CalCheck Vitamin D total II test material is stored at 2-8°C. The CalChecks are tested at T=0 and at specified intervals over the shelf life of the device up to the planned shelf life plus one month. For the lot stability, data for the time-points at 0, 7, 13 and 19 months tested in duplicate will be available. The average on-test recovery value will be calculated as percent recovery compared to the unstressed reference value (stored at -20°C).
4.17. Sample Stability
No new data was collected using the Elecsys Vitamin D total II assay. Sample stability data generated with the Elecsys Vitamin D Assay (K113546) is applicable since the claims and sample materials did not change (sample stability is not dependent on assay reagent formulation).
EXTERNAL (CLINICAL) TESTING 5.
Clinical samples were collected at three US sites in order to establish the reference range values for the Elecsys Vitamin D total II assay. Serum samples were prospectively collected from approximately 200 apparently healthy adults at each of the three locations (northern site, midwest site and southern site) during summer and winter seasons (totaling approximately 600 Study Subjects). Study subjects were pre-screened to exclude subjects taking supplements or various drugs, those who were over or under weight, those with a medical history that might suggest variations in bone health, and pregnant and lactating women. Conditions such as parathyroid or thyroid disorders, and kidney disease, as well as electrolyte composition were determined by testing the samples. Those samples with properties outside of the published reference range for the particular test were excluded from the calculation of the Elecsys Vitamin D total II reference range study. They were assayed with Elecsys Vitamin D total II on the cobas e 411 immunoassay analyzer.
The Reference Interval for the total population of the Elecsys Vitamin D total II assay is 7.61 - 55.5 ng/mL. The established reference ranges for Elecsys Vitamin D total II are provided below in Table 5.
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| N | Mean | Median | 2.5th Percentile(90 % CIª) | 97.5th Percentile(90 % CI) | Units |
|---|---|---|---|---|---|
| 421 | 25.8 | 23.6 | 7.61(6.11, 8.44) | 55.5(52.4, 64.5) | ng/mL |
| Table 5: Results of 2.5th and 97.5th Percentiles for Elecsys Vitamin D total II | ||
|---|---|---|
| -- | --------------------------------------------------------------------------------- | -- |
a) Confidence interval
CONCLUSIONS 6.
The information provided in this 510(k) Premarket Notification will support a determination of substantial equivalence for Elecsys Vitamin D total II, CalSet Vitamin D total II, PreciControl Vitamin D total II and CalCheck Vitamin D total II. The data supports a safe, effective device which performs as well or better than the predicate devices.
§ 862.1825 Vitamin D test system.
(a)
Identification. A vitamin D test system is a device intended for use in clinical laboratories for the quantitative determination of 25-hydroxyvitamin D (25-OH-D) and other hydroxylated metabolites of vitamin D in serum or plasma to be used in the assessment of vitamin D sufficiency.(b)
Classification. Class II (special controls). Vitamin D test systems must comply with the following special controls:(1) Labeling in conformance with 21 CFR 809.10 and
(2) Compliance with existing standards of the National Committee on Clinical Laboratory Standards.