ADVIA Centaur Vitamin D Total (VitD)

K200509 · Siemens Healthcare Diagnostics, Inc. · MRG · May 29, 2020 · Clinical Chemistry

Device Facts

• Record ID: K200509
• Device Name: ADVIA Centaur Vitamin D Total (VitD)
• Applicant: Siemens Healthcare Diagnostics, Inc.
• Product Code: MRG · Clinical Chemistry
• Decision Date: May 29, 2020
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.1825
• Device Class: Class 2
• Attributes: Pediatric

Intended Use

The ADVIA Centaur® Vitamin D Total (VitD) assay is for in vitro diagnostic use in the quantitative determination of total 25(OH)vitamin D in human serum and plasma (EDTA, lithium heparin, sodium heparin) using the ADVIA Centaur® systems. The ADVIA Centaur® VitD assay is intended as an aid in the determination of vitamin D sufficiency.

Device Story

ADVIA Centaur® Vitamin D Total assay is an 18-minute competitive chemiluminescent immunoassay; utilizes anti-25(OH)vitamin D monoclonal mouse antibody labeled with acridinium ester, anti-fluorescein monoclonal mouse antibody bound to paramagnetic particles, and fluorescein-labeled vitamin D analog. Input: human serum or plasma (EDTA, lithium heparin, sodium heparin). Operation: inverse relationship between sample 25(OH)vitamin D concentration and detected relative light units (RLU). Performed on ADVIA Centaur® XP analyzers in clinical laboratory settings by trained personnel. Output: quantitative 25(OH)vitamin D concentration (ng/mL). Results aid clinicians in determining vitamin D sufficiency status; supports patient management regarding vitamin D deficiency/insufficiency.

Clinical Evidence

Bench testing only. Precision evaluated over 20 days (n=80 replicates/sample); total CVs 3.6%–9.6%. Linearity confirmed 2.78–151.00 ng/mL. LoQ 4.2 ng/mL. Method comparison (n=126) against predicate showed R=0.99, slope 1.03–1.04. Matrix comparison (n=74) confirmed equivalence across serum and plasma types (R=0.99). Reference interval verification performed on 20 healthy individuals.

Technological Characteristics

Competitive chemiluminescent immunoassay. Reagents: monoclonal mouse anti-VitD (acridinium ester labeled), anti-fluorescein monoclonal mouse antibody on paramagnetic particles, vitamin D-analog fluorescein conjugate. Energy source: ADVIA Centaur system instrumentation. Connectivity: System-integrated. Sterilization: Not applicable (reagent kit).

Indications for Use

Indicated for quantitative determination of total 25(OH)vitamin D in human serum and plasma to aid in assessing vitamin D sufficiency in adults and pediatric patients (12 months to 21 years).

Regulatory Classification

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.

Special Controls

*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.

Predicate Devices

• ADVIA Centaur® Vitamin D Total (VitD) assay (K133156)

Submission Summary (Full Text)

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FDA U.S. FOOD & DRUG ADMINISTRATION

# 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY

ASSAY ONLY

## I Background Information:

A 510(k) Number

K200509

B Applicant

Siemens Healthcare Diagnostics Inc.

C Proprietary and Established Names

ADVIA Centaur® Vitamin D Total (VitD)

D Regulatory Information

| Product Code(s) | Classification | Regulation Section | Panel |
| --- | --- | --- | --- |
| MRG | Class II | 21 CFR 862.1825 - Vitamin D Test System | CH - Clinical Chemistry |

## II Submission/Device Overview:

A Purpose for Submission:

Modified device to reinstate plasma (sodium heparin, lithium heparin, K₂ EDTA, K₃ EDTA) as a sample matrix.

B Measurand:

25-hydroxyvitamin D

C Type of Test:

Quantitative chemiluminescent

Food and Drug Administration

10903 New Hampshire Avenue

Silver Spring, MD 20993-0002

www.fda.gov

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# III Intended Use/Indications for Use:

## A Intended Use(s):

See Indications for Use below.

## B Indication(s) for Use:

The ADVIA Centaur® Vitamin D Total (VitD) assay is for in vitro diagnostic use in the quantitative determination of total 25(OH)vitamin D in human serum and plasma (EDTA, lithium heparin, sodium heparin) using ADVIA Centaur® analyzers. The ADVIA Centaur® VitD assay is intended as an aid in the determination of vitamin D sufficiency.

## C Special Conditions for Use Statement(s):

Rx - For Prescription Use Only

## D Special Instrument Requirements:

ADVIA Centaur® XP analyzer

# IV Device/System Characteristics:

## A Device Description:

The ADVIA Centaur® Vitamin D reagent kit comes in two configurations (100 or 500 test kit) and each kit contains the following:

- ReadyPack® primary reagent pack containing ADVIA Centaur Vit D Lite Reagent, Solid Phase Reagent, and Ancillary Well Reagent
- ReadyPack® ancillary pack containing ADVIA Centaur Vit D Ancillary Reagent
- ADVIA Centaur® Vit D Low Calibrator
- ADVIA Centaur® Vit D High Calibrator
- ADVIA Centaur® systems Vit D Master Curve card
- ADVIA Centaur® systems Vit D Calibrator Assigned Value Card

The Vit D Reagents consists of the following:

### Lite Reagent 5.0 mL/reagent pack:

The reagent contains anti-Vit D (monoclonal mouse) antibody labeled with acridinium ester (~0.8 µg/mL) in buffer with bovine serum albumin, mouse IgG, and sodium azide (< 0.1%)

### Solid Phase Reagent 10.0 mL/reagent pack:

The Solid Phase Reagent contains anti-fluorescein (monoclonal mouse)-coated paramagnetic particles (PMP) (~0.60 mg/mL) in buffer with bovine serum albumin, surfactant, and sodium azide (< 0.1%)

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Ancillary Well Reagent 5.0 mL/reagent pack:
The Ancillary Well Reagent contains vitamin D-analog conjugated to fluorescein (~0.2 µg/mL) and 1-anilinonaphthalene-8-sulfonic acid in buffer with bovine serum albumin and sodium azide (< 0.1%)

Ancillary Reagent Pack 25.0 mL/reagent pack:
The Ancillary Reagent Pack contains releasing agent in buffered saline with sodium azide (<0.1%) and stabilizers

Material Required but Not Provided
- ADVIA Centaur® Wash 1

Optional Materials
- ADVIA Centaur® Vit D Quality Control Material
- ADVIA Centaur® Vit D Diluent
- ADVIA Centaur® Vit D Master Curve Material (MCM)

B Principle of Operation:
The ADVIA Centaur® Vit D assay is an 18-minute antibody competitive immunoassay that uses an anti-fluorescein monoclonal mouse antibody covalently bound to paramagnetic particles (PMP), an anti-25(OH)vitamin D monoclonal mouse antibody labeled with acridinium ester (AE), and a vitamin D analog labeled with fluorescein.

An inverse relationship exists between the total amount of 25(OH) vitamin D present in the patient sample and the amount of relative light units (RLU) detected by the system.

V Substantial Equivalence Information:

A Predicate Device Name(s):
ADVIA Centaur® Vitamin D Total (VitD) Assay

B Predicate 510(k) Number(s):
K133156

C Comparison with Predicate(s):

| Device & Predicate Device(s): | K200509 | K133156 |
| --- | --- | --- |
| Device Trade Name | ADVIA Centaur® Vitamin D Total (Vit D) assay | Same |
| General Device Characteristic Similarities | | |

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| Device & Predicate Device(s): | K200509 | K133156 |
| --- | --- | --- |
| Intended Use/Indications For Use | For the quantitative determination of total 25(OH)vitamin D. The assay is intended as an aid in the determination of vitamin D sufficiency. | Same |
| Measurement | Quantitative | Same |
| Assay Principle | Competitive immunoassay | Same |
| Technology | Chemiluminescence | Same |
| Detection Antibody | Monoclonal mouse antibody labeled with acridium ester (AE) | Same |
| Capture Antibody | Anti-fluorescein labeled (FITC) monoclonal mouse antibody covalently bound to paramagnetic particles (PMP). | Same |
| Measuring Range | 4.2 to 150 ng/mL | Same |
| General Device Characteristic Differences | | |
| Ancillary Pack Reagent | Buffering agent (high molarity) and enhanced releasing agent (low molecular weight) | Buffering agent (low molarity) and releasing agent (high molecular weight) |

VI Standards/Guidance Documents Referenced:

- CLSI. EP17-A2, 2012 Evaluation of Detection Capability for Clinical Laboratory Measurement Procedures; Approved Guideline—Second Edition
- CLSI EP06-A, 2003 Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach; Approved Guideline – First Edition
- CLSI EP05-A3, 2014; Evaluation of Precision of Quantitative Measurement Procedures; Approved Guideline—Third Edition
- CLSI EP34, 2018; Establishing and Verifying an Extended Measuring Interval Through Specimen Dilution and Spiking – First Edition
- CLSI EP28-A3c – formerly C28-A3c, 2010; Defining, Establishing and Verifying

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Reference Intervals in the Clinical Laboratory; Approved Guideline – Third Edition

- CLSI EP25-A, 2009; Evaluation of Stability of In Vitro Diagnostic Reagents; Approved Guideline– First Edition

## VII Performance Characteristics (if/when applicable):

### A Analytical Performance:

#### 1. Precision/Reproducibility:

Within run and total imprecision were evaluated by analyzing six serum samples at the concentrations below. Each sample was assayed in 2 replicates per run, 2 runs per day for 20 days for a total of 80 replicates per sample. Testing was performed with one reagent lot on one ADVIA Centaur® XP analyzer. Results are summarized below:

| Sample | Mean (ng/mL) | Repeatability | | Within-Lab | |
| --- | --- | --- | --- | --- | --- |
| | | SD | % CV | SD | % CV |
| Serum 1 | 21.29 | 1.36 | 6.4 | 2.04 | 9.6 |
| Serum 2 | 26.10 | 1.56 | 6.0 | 2.37 | 9.1 |
| Serum 3 | 32.16 | 1.71 | 5.3 | 2.38 | 7.4 |
| Serum 4 | 65.47 | 2.52 | 3.8 | 3.60 | 5.5 |
| Serum 5 | 84.12 | 1.90 | 2.3 | 3.34 | 4.0 |
| Serum 6 | 132.32 | 3.13 | 2.4 | 4.76 | 3.6 |

#### 2. Linearity:

This study was conducted following the guidelines of CLSI EP06-A: Evaluation of the Linearity of Quantitative Measurement Procedures.

A high (>150 ng/mL) Vitamin D sample was made by spiking 25(OH) Vitamin D into a human serum sample. Vitamin D stripped human serum was used as the low sample. A dilution series was then made by mixing the high and low samples to encompass the measuring interval of the Vitamin D assay. Each resulting sample was run in triplicate on one reagent lot on one ADVIA Centaur® XP analyzer.

The sponsor concluded that the modified assay is linear from 2.78 ng/mL to 151.00 ng/mL on the ADVIA Centaur® XP analyzer. A measuring range from 4.2 – 150 ng/mL is claimed in the labeling.

### Dilution Recovery:

To validate the labeling instructions that patient samples with Vitamin D levels above the claimed measuring range can be manually or automatically diluted 1:2, 5 unique serum samples were spiked with 25(OH) Vitamin D to produce concentrations ranging from 186 - 211 ng/mL. The samples were then diluted 1:2 with ADVIA Centaur® VitD diluent and assayed for recovery. The recoveries ranged from 97% to 109%.

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| Sample | Dilution | Observed (ng/mL) | Expected (ng/mL) | Recovery % |
| --- | --- | --- | --- | --- |
| 1 | 1:2 | 96.0 | 97.5 | 98 |
| 2 | 1:2 | 96.3 | 93.0 | 103 |
| 3 | 1:2 | 102.6 | 103.5 | 99 |
| 4 | 1:2 | 102.5 | 105.5 | 97 |
| 5 | 1:2 | 102.1 | 93.5 | 109 |

3. Analytical Specificity/Interference:

Assay specificity was established during the review of k133156.

4. Assay Reportable Range:

The ADVIA Centaur® VitD assay measures 25(OH) vitamin D from concentrations of 4.2 to 150 ng/mL (10.5 to 375 nmol/L). The claimed measuring range is defined by the limit of quantitation (LoQ), linearity, and method comparison studies.

5. Traceability, Stability, Expected Values (Controls, Calibrators, or Methods):

The ADVIA Centaur® VitD calibrators are traceable to the NIST SRM972a 25-OH Vitamin D reference material.

6. Detection Limit:

A limit of blank (LoB) study was conducted following the guidelines of CLSI EP17-A2: Evaluation of Detection Capability for Clinical Laboratory Measurement Procedures. Four blank samples with no Vitamin D were run n=6 for 6 days, 2 runs/day on 2 reagent lots for a total n=72 reps per sample per lot, on one ADVIA Centaur® XP analyzer. The LoB was calculated as 1.52 ng/mL.

A limit of detection (LoD) study was conducted following the guidelines of CLSI EP17-A2. Four (4) low level samples were made by diluting individual patient samples with Vitamin D stripped serum to concentrations around the expected LoD. Samples were run n=6 for 5 days, 2 runs per day on 2 reagent lots on one ADVIA Centaur® XP analyzer for a total n=60 replicates per sample, and lot. The LoD was calculated as 3.11 ng/mL.

A limit of quantitation (LoQ) study was conducted following the guidelines of CLSI EP17-A2. Thirteen (13) unique native serum specimens were diluted with Vitamin D stripped serum to produce concentrations ranging from 1.24 to 25.6 ng/mL. Samples were run n=6 for 5 days, 2 runs per day on two reagent lots, on one ADVIA Centaur® XP analyzer for a total n=60 replicates per sample, lot, platform, across 5 days. Data was analyzed 2 point using a stored calibration. LoQ was calculated by precision profile and was defined as the concentration at which the within-lab coefficient of variation was 20%. The LoQ was calculated as 4.24 ng/mL.

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These studies support the following labeling claims:

| Limit of blank | 1.7 ng/mL |
| --- | --- |
| Limit of detection | 3.2 ng/mL |
| Limit of quantitation | 4.2 ng/mL |

# 7. Assay Cut-Off:

Not applicable.

# B Comparison Studies:

# 1. Method Comparison with Predicate Device:

To compare results from the modified device to the predicate, one hundred twenty-six (126) serum samples (including 118 native and 8 contrived) were run in singlicate over 2 days using two lots of modified reagent and compared to the predicate assay on one ADVIA Centaur® XP analyzer. Contrived samples were made by spiking 25(OH) Vitamin D into individual samples to obtain Vitamin D levels at the high end of the Vitamin D assay measuring range. Results using Deming regression are summarized below:

| Reagent Lot (y) | Reagent Lot 1 | Reagent Lot 2 |
| --- | --- | --- |
| N | 126 | 126 |
| Range (ng/mL) | 5.93 - 130.85 | 5.93 - 130.85 |
| Intercept | 0.85 | 1.74 |
| Slope | 1.03 | 1.04 |
| R Value | 0.99 | 0.99 |

# 2. Matrix Comparison:

To validate use of the ADVIA Centaur® VitD assay with different claimed sample types, the sponsor evaluated 66 native and 8 contrived matched sample sets (serum, serum separator tube, lithium heparin, sodium heparin, K2 EDTA, K3 EDTA) ranging in concentration from 13.0 to $142.9\mathrm{ng / mL}$ . Samples were run along with calibrators and control on one ADVIA Centaur® XP analyzer with one lot of reagent. All samples were tested in replicates of 3 and the first replicate was used to perform a Weighted-Deming fit regression analysis comparing all tube types to serum collected in a plain tube without a serum separator. Pearson correlation coefficient (R) was calculated and the results are summarized below:

| Tube Type | Slope | Intercept | R |
| --- | --- | --- | --- |
| Serum Separator | 0.97 | 0.87 | 0.99 |
| Sodium Heparin | 1.02 | -1.13 | 0.99 |
| Lithium Heparin | 0.99 | -0.18 | 0.99 |
| K2EDTA | 0.97 | 0.64 | 0.99 |
| K3EDTA | 0.96 | 0.68 | 0.99 |

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The results of this study support the use of the ADVIA Centaur® Vitamin D Total (VitD) assay with venous blood collected in serum tubes with or without a separator as well as plasma from venous blood collected in tubes with the following anticoagulants: sodium heparin, lithium heparin, K₂ EDTA, and K₃ EDTA.

## C Clinical Studies:

1. Clinical Sensitivity:
Not applicable

2. Clinical Specificity:
Not applicable

3. Other Clinical Supportive Data (When 1. and 2. Are Not Applicable):
Not applicable

## D Clinical Cut-Off:

Not applicable

## E Expected Values/Reference Range:

The sponsor performed a study referencing CLSI EP28-A3c: Defining, Establishing, and Verifying Reference Intervals in the Clinical Laboratory to verify that the existing Reference Interval could be transferred to the modified Vitamin D assay. Twenty (20) samples from apparently healthy individuals were analyzed on one ADVIA Centaur® XP analyzer using one lot of the modified reagent. Results of the verification study and the performance studies support that the reference interval could be transferred to the modified reagent.

| | Adults | Pediatric (12 months up to 21 years) |
| --- | --- | --- |
| Median | 22.5 ng/mL | 23.8 ng/mL |
| Observed Range 2.5^{th} to 97.5^{th} Percentile | 7.4 – 44.0 ng/mL | 11.4 – 45.8 ng/mL |

In addition, the labeling contains the following information on Vitamin D levels, and which is drawn from literature references:

| Vitamin D Status | Range, Adult ng/mL (nmol/L) | Range, Pediatric ng/mL (nmol/L) |
| --- | --- | --- |
| Deficiency | < 20 ng/mL (50 nmol/L) | < 15 ng/mL (37.5 nmol/L) |
| Insufficiency | 20– < 30 ng/mL (50– < 75 nmol/L) | 15 – < 20 ng/mL (37.5– < 50 nmol/L) |

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| Vitamin D Status | Range, Adult ng/mL (nmol/L) | Range, Pediatric ng/mL (nmol/L) |
| --- | --- | --- |
| Sufficiency | 30–100 ng/mL (75–250 nmol/L) | 20–100 ng/mL (50–250 nmol/L) |

Holick MF. Vitamin D Deficiency. N Engl J Med. 2007;357:266–81.

Holick MF. MrOs is D-ficient. J Clin Endocrinol Metab. 2009;94(4):1092–3.

Rollins G. Vitamin D Testing–What’s the Right Answer? Labs Grapple with Confusing Analytics, Evidence. Clinical Laboratory News. July 2009;35(7): 1,6.

Freeman R. Vitamin D: The sunshine hormone. How and when to treat deficiencies. Menopausal Medicine. May 2009; S8–11.

Misra M, Pacaud D, Petryk A, Collett-Solberg PF, Kappy M. Vitamin D Deficiency in Children and Its Management: Review of Current Knowledge and Recommendations. Pediatrics. 2008;122:398–417

## VIII Proposed Labeling:

The labeling supports the finding of substantial equivalence for this device.

## IX Conclusion:

The submitted information in this premarket notification is complete and supports a substantial equivalence decision.

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