The Diazyme 25-OH Vitamin D EIA Kit is designed for the quantification of total 25-OH Vitamin D in human serum and plasma. The assay results are to be used in parallel with other clinical data to assess the Vitamin D status of a patient. For in vitro diagnostic use only.

The 25-OH Vitamin D EIA Control Kit is intended for use as quality controls for the Diazyme 25-OH Vitamin D EIA Kit only. For in vitro diagnostic use only.

Device Description

The Diazyme 25-hydroxy (25-OH) Vitamin D EIA uses the enzyme immuno-assay technology (EIA). It is based on the competition (for a 25-OH Vitamin D antibody) between a 25-OH Vitamin D conjugate coated on a microplate and the 25-OH Vitamin D content of a serum sample. Vitamin D samples are first extracted for 15 min at room temperature using reagent EX and deep-well pre-dilution strips. The extracted Vitamin D samples are then transferred to the coated microplate wells and the addition of reagent R1 (containing an antibody for 25-OH Vitamin D) allows for the competition to proceed. After a first incubation, the microplate is washed and reagent R2 (HRP-labeled secondary antibody) is added. Following a second incubation and a washing step, reagent R3 (TMB substrate) is added. After a final incubation step, the reaction is stopped by adding the STOP solution and the colorimetric signal of the microplate is measured at 450 nm (primary wavelength) to which the background signal can be subtracted by using a secondary wavelengths (620 to 650 nm). The 25-OH Vitamin D concentration of a patient sample is inversely proportional to the measured absorbance at 450 nm. The whole assay procedure takes about two hours. 25-OH Vitamin D EIA calibrator set is intended for use as a calibration for the Diazyme 25-OH Vitamin D EIA kit only. Six calibration levels are needed for each run. Calibrators are treated exactly the same as patient samples.

25-OH Vitamin D EIA 2-point control kit is intended for use with the Diazyme 25-OH Vitamin D EIA kit only. Controls are treated exactly the same as patient samples. The quality controls assist laboratory users in verification steps ensuring that the assay reagents are functioning correctly. Users are instructed to verify the calibration curve with the controls.

AI/ML Overview

Here's an analysis of the provided 510(k) summary, specifically focusing on the acceptance criteria and study details for the Diazyme 25-OH Vitamin D EIA Kit:

Introduction
The Diazyme 25-OH Vitamin D EIA Kit is designed for the quantification of total 25-OH Vitamin D in human serum and plasma, with results used alongside other clinical data to assess Vitamin D status. The following sections detail the performance criteria and the studies conducted to demonstrate its effectiveness.

1. Table of Acceptance Criteria and Reported Device Performance

Performance Metric	Acceptance Criteria (Implied/Standard)	Reported Device Performance (Diazyme 25-OH Vitamin D EIA Kit)
Precision	CV% typically < 10-15% for clinical assays, varying by concentration.	Within-run CV: 2.2% - 13.3%
		Total CV: 4.9% - 17.6%
	(Specifically, per CLSI EP5-A guideline)	(See Table in Section 4 for detailed values per sample)
Linearity/Reportable Range	Assay should be linear within its claimed measuring range.	Linear Range: 8.3 ng/mL to 143.6 ng/mL
	(Per CLSI EP6-A guideline)	(Found to be linear between 6.7 ng/mL and 143.6 ng/mL, but adjusted to LoQ.)
Limit of Blank (LoB)	Determined according to CLSI EP17-A.	LoB: 3.0 ng/mL
Limit of Detection (LoD)	Determined according to CLSI EP17-A.	LoD: 5.6 ng/mL
Limit of Quantitation (LoQ)	Determined according to CLSI EP17-A.	LoQ: 8.3 ng/mL
Interference Study	Less than 10% deviation from expected values with interfering substances.	Less than 10% deviation for listed substances:
	(Per CLSI EP7-A2 protocol)	- Ascorbic Acid (10 mM)
		- Free Bilirubin (40 mg/dL)
		- Conjugated Bilirubin (40 mg/dL)
		- Triglyceride (450 mg/dL)
		- Hemoglobin (500 mg/dL)
Cross-Reactivity	Minimal or acceptable cross-reactivity with related substances.	- 25-OH Vitamin D3: 100.0%
		- 25-OH Vitamin D2: 96.8%
		- Vitamin D3: -0.3%
		- Vitamin D2: -3.3%
		- 1,25-OH Vitamin D3: 94.8% (but does not interfere up to 100X normal levels)
		- 1,25-OH Vitamin D2: 59.7% (but does not interfere up to 100X normal levels)
		- 24R,25-OH Vitamin D3: 23.9% (but does not interfere up to 100X normal levels)
		- 3-epi-25-OH Vitamin D3: 84.9%
		- 3-epi-25-OH Vitamin D2: 67.9%
Method Comparison (vs. Predicate)	Good correlation with predicate device (e.g., R-value > 0.9).	n: 58
		Slope: 0.941
		Intercept: +1.448
		Correlation coefficient: 0.930
		Range of values: 11.9 ng/mL - 131.5 ng/mL
Matrix Comparison (Serum vs. Plasma)	Good agreement (e.g., R^2 > 0.95, slope near 1, intercept near 0).	Li-Heparin plasma vs. Serum: y = 0.993x + 1.855, R^2 = 0.961
		K3-EDTA plasma vs. Serum: y = 1.006x + 2.901, R^2 = 0.967
Reference Range Study	Establish a representative reference range for a healthy population.	n: 157
		Lowest Conc: 8.4 ng/mL
		Highest Conc: 61.3 ng/mL
		Median Conc: 29.1 ng/mL
		Observed Range (2.5th-97.5th percentile): 12.0 to 55.0 ng/mL

2. Sample Sizes Used for the Test Set and Data Provenance

Precision Study:
- Sample Size: 10 precision levels (2 serum controls, 8 serum samples). Each specimen had 80 measurements in total (20 days * 4 measurements/day). So, 10 samples * 80 measurements each = 800 individual data points.
- Data Provenance: Not explicitly stated, but derived from internal laboratory testing using serum controls and serum samples. Likely prospective internal testing.
Linearity/Reportable Range:
- Sample Size: 11 levels of linearity prepared from a high and low serum sample. Each level was tested in triplicates.
- Data Provenance: Not explicitly stated, but derived from internal laboratory testing using spiked serum samples. Likely prospective internal testing.
LoB/LoD/LoQ:
- Sample Size: Not explicitly detailed beyond stating the methodology. This typically involves multiple replicates of blank, low-concentration, and representative samples over several runs.
- Data Provenance: Not explicitly stated, but derived from internal laboratory testing. Likely prospective internal testing.
Interference Study:
- Sample Size: Not explicitly stated for the number of tested samples, but experiments focused on a set of common interfering substances.
- Data Provenance: Not explicitly stated, but derived from internal laboratory testing. Likely prospective internal testing using spiked samples.
Cross-Reactivity Study:
- Sample Size: Not explicitly detailed for the number of replicates or unique samples, but involved testing specific Vitamin D metabolites.
- Data Provenance: Not explicitly stated, but derived from internal laboratory testing using serum pool samples spiked with various cross-reactants. Likely prospective internal testing.
Method Comparison:
- Sample Size: 58 human serum samples (7 of which were spiked).
- Data Provenance: Human serum samples. The origin (country) is not specified, but the testing was internal. The nature suggests these were likely prospective or retrospectively collected clinical samples used in a prospective comparison study.
Matrix Comparison:
- Sample Size: 66 matched sets of serum, K3-EDTA plasma, and Li-Heparin plasma. Includes seven spiked patient samples.
- Data Provenance: Matched human samples in different matrices. The origin (country) is not specified. Likely prospective or retrospectively collected clinical samples used in a prospective comparison study.
Reference Range Study:
- Sample Size: 157 apparently healthy individuals.
- Data Provenance: Samples from three different US geographical locations: 47 from Pennsylvania (Northern U.S.), 56 from Tennessee (Central U.S.), and 54 from Texas (Southern U.S.). These were obtained from certified commercial sources (ProMedDx, LLC and Dx Biosamples) or collected under an IRB approved protocol. This indicates retrospective collection of samples from defined populations.

3. Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications of Those Experts

This type of in-vitro diagnostic device (EIA kit) does not typically involve human experts for establishing ground truth in the way AI/imaging devices do (e.g., radiologists reviewing images). Instead, the "ground truth" or reference values are established by:

Reference Methods: For method comparison, the predicate device (IDS 25-OH Vitamin D EIA, K021163) serves as the reference against which the new device is compared. This predicate device itself would have undergone its own rigorous validation.
Known Concentrations: For studies like linearity, LoB/LoD/LoQ, interference, and cross-reactivity, ground truth is established by preparing samples with known, precisely measured concentrations of the analyte or interfering/cross-reacting substances. This is done in a laboratory setting using calibrated equipment and certified reference materials.
Clinical Standards: For the reference range study, the ground truth is the measured 25-OH Vitamin D concentration in a statistically representative healthy population, measured by the Diazyme device itself.

Therefore, the concept of "number of experts" adjudicating ground truth in the context of clinical pathology or imaging is not applicable here. The expertise lies in the rigorous adherence to established laboratory guidelines (CLSI protocols) and the use of reference methods/materials.

4. Adjudication Method for the Test Set

Again, for an in-vitro diagnostic assay measuring an analyte, adjudication methods like N+1 or 2+1 (common in AI imaging) are not applicable. The "adjudication" is inherent in the analytical process:

Predicate Device: For method comparison, the predicate device's measurement serves as the standard.
Statistical Analysis: Results are compared using statistical methods (e.g., linear regression, correlation coefficient) to determine agreement and equivalence, based on predefined statistical acceptance criteria.
CLSI Guidelines: Adherence to CLSI (Clinical and Laboratory Standards Institute) protocols ensures standardized and robust evaluation.

5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study

No, an MRMC comparative effectiveness study was not done.
This type of study is primarily relevant for diagnostic imaging or interpretation tasks where multiple human readers interpret cases, and AI assistance might influence their performance.
The Diazyme 25-OH Vitamin D EIA Kit is an automated laboratory test for quantitative measurement of an analyte, not an interpretation task performed by human readers.

6. Standalone Performance (Algorithm Only Without Human-in-the-Loop Performance)

Yes, standalone performance was done.
The performance metrics reported (precision, linearity, LoB/LoD/LoQ, interference, cross-reactivity, and method/matrix comparison) all represent the performance of the Diazyme 25-OH Vitamin D EIA Kit itself, as an "algorithm only" in the sense of an automated test.
There is no "human-in-the-loop" interaction for interpreting or modifying the test results generated by the EIA kit; the results are directly quantitative measurements.

7. Type of Ground Truth Used

The ground truth used depends on the specific study:

Known concentrations/Reference materials: For precision, linearity, LoB/LoD/LoQ, interference, and cross-reactivity studies. These are precisely prepared samples with a known amount of the analyte or interfering substance.
Comparison to a Legally Marketed Predicate Device: For the method comparison study, the measurements obtained by the predicate device (IDS 25-OH Vitamin D EIA, K021163) served as the comparative ground truth.
Patient Outcome Data: Not directly used as ground truth for performance validation, although the assay results are intended to be used in parallel with other clinical data to assess Vitamin D status, which could indirectly relate to patient outcomes. However, the regulatory submission focuses on analytical performance rather than direct impact on patient outcomes.

8. Sample Size for the Training Set

This device is an EIA kit, which is a chemical assay, not an AI/machine learning algorithm that requires a "training set" in the conventional computational sense.
Therefore, the concept of a "training set" is not applicable here. The development of the assay's reagents and protocols would involve optimization and piloting, but not an ML-style training dataset.

9. How the Ground Truth for the Training Set Was Established

As the concept of a training set is not applicable, the method of establishing its ground truth is also not applicable.
The "ground truth" during the development and optimization of such an assay would be based on established biochemical principles, spiked samples with known concentrations, and comparisons to established gold-standard methods or reference laboratories during the R&D phase.

Summary

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122420

510(k) SUMMARY

NOV 2 8 2012

.This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of SMDA 1990 and 21 CFR 807.92

Submitter's name:

Diazyme Laboratories

12889 Gregg Court

Poway, CA 92064

Submitter's address:

Name of Contact Person:

Dr. Abhijit Datta Diazyme Laboratories 12889 Gregg Court Poway, CA 92064 Phone: 858-455-4762 Fax: 858-455-2120

Date the Summary was Prepared:

Name of the Device

October 17, 2012

25-OH Vitamin D EIA Kit 25-OH Vitamin D EIA Control Kit

Trade Name:

Common/Usual Name

Device Classification Name

Product code:

Panel: ﭘ

... .. ..

Submission Type

Regulation Number

25-OH Vitamin D EIA Kit 25-OH Vitamin D EIA Control Kit

Vitamin D Assay

Vitamin D Test System

MRG - Vitamin D Test System

JJX - Single (specified) Analyte Controls (Assayed and Unassayed)

Chemistry (75)

510k

21 CFR 862.1825 - Vitamin D Test System

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21 CFR 862.1660 - Quality Control material (Assayed and Un-assayed)

Device Class

Predicate Device:

Manufacturing Address

II (Assay) I (Control)

The 25-OH Vitamin D EIA Test Kit and Control Kit is substantially equivalent to the currently marketed 25-OH Vitamin D EIA (K021163).

Diazyme Laboratories 12889 Gregg Court Poway, CA 92064 USA

Establishment Registration .

2032900

DESCRIPTION OF THE DEVICE

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INDICATIONS FOR USE

The 25-OH Vitamin D EIA Control Kit is intended for use as quality controls for the Diazyme 25-OH Vitamin D EIA Kit only. For in vitro diagnostic use only.

Table 1 Summary of Assay Kit Components

-IDS 25-OH Vitamin D EIA(predicate K021163)	Diazyme 25-OH Vitamin D EIA
Kit can be only used for the 25-OH Vitamin D quantification on microplate readers.	Kit can be only used for the 25-OH Vitamin D quantification on microplate readers
Antibody Coated Plate (MICROPLAT)	Coated Microplate
Microplate with 25-OH Vitamin D sheep polyclonal antibody linked to the inner surface of the polystyrene wells, 12 x 8 well strips in a foil pouch with desiccants.	Microplate with a 25-OH Vitamin D derivative linked to the inner surface of the polystyrene wells, 12 x 8 well strips in a foil pouch with desiccants.
Adhesive plate sealer 8 per kit	N/A
Dissociation buffer (25-D Biotin)1 bottle Proprietary reagent for dissociating Vitamin D 25-OH Vitamin D labeled with Biotin Stabilizers	Extraction solution (EX)1 bottle Extraction solution containing organic solvents.
Wash Buffer (WASHBUF)1 bottle Phosphate buffered saline containing Tween	Wash Buffer (Wash 20X)1 bottle A 20X concentrate of a phosphate buffered saline containing Tween 20.
Enzyme Conjugate (ENZYMCONJ)1 bottle Phosphate buffered saline containing avidin linked to horseradish peroxidase, protein, enzyme stabilizers and preservatives.	Reagent 11 bottle Phosphate buffered saline containing a sheep monoclonal antibody specific to 25-OH Vitamin D, stabilizers and preservatives.
N/A	Reagent 21 bottle Phosphate buffered saline containing an anti-sheep IgG linked to horseradish peroxidase, stabilizers and preservatives.
TMB substrate (SUBS)1 bottleProprietary aqueous formulation of tetra-methylbenzidine (TMB) and hydrogenperoxide.	Reagent 31 bottleProprietary aqueous formulation of tetra-methylbenzidine (TMB) and hydrogenperoxide.
STOP solution (HCl)1 bottleConcentrated solution of hydrochloric acidHCl.	STOP solution (HCl)1 bottleConcentrated solution of hydrochloric acidHCl.
Calibrator set1 x 1.0 mL Calibrator 01 x 1.0 mL Calibrator 11 x 1.0 mL Calibrator 21 x 1.0 mL Calibrator 31 x 1.0 mL Calibrator 41 x 1.0 mL Calibrator 51 x 1.0 mL Calibrator 6	Calibrator set1 x 1.0 mL Calibrator 11 x 1.0 mL Calibrator 21 x 1.0 mL Calibrator 31 x 1.0 mL Calibrator 41 x 1.0 mL Calibrator 51 x 1.0 mL Calibrator 6
Control set1 x 1.0mL Control 11 x 1.0mL Control 2	Control set1 x 1.0mL Control 11 x 1.0mL Control 2

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PERFORMANCE TESTING SUMMARIES ON DYNEX MICROPLATE READER

Precision Study

The precision of the Diazyme 25-OH Vitamin D EIA was evaluated according to the Clinical and Laboratory Standards Institute (CLSI) EP5-A guideline.

The precision of the Diazyme 25-OH Vitamin D Enzyme-Immuno-Assay (25-OH Vitamin D EIA) was evaluated according to the Clinical and Laboratory Standards Institute (CLSI) EPS-A guideline. A total of 10 precision levels were used in the study:

Two serum controls (containing 26.0 ng/mL and 52.3 ng/mL). �
Eight serum samples distributed across the dynamic range of the assay. .

Controls and samples were measured daily, over the span of 20 days. Each day, a set of four measurements was obtained for each specimen with two independent runs, each run being done in duplicates.

The mean value (Mean), standard deviation, within-run imprecision and total imprecision are calculated and summarized in the following tables:

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Specimen	n	Mean (ng/mL)	Within-run SD (ng/mL)	Within-run CV (%)	Total SD (ng/mL)	Total CV (%)
Control #1	80	26.0	0.97	3.7	1.55	6.0
Control #2	80	52.3	1.43	2.7	3.93	7.5
Sample #1	80	23.3	1.65	7.1	1.89	8.1
Sample #2	80	40.0	3.70	9.2	3.75	9.4
Sample #3	80	52.8	1.18	2.2	4.03	7.6
Sample #4	80	69.3	1.94	2.8	6.03	8.7
Sample #5	80	84.1	4.69	5.6	6.33	7.5
Sample #6	80	101.4	4.65	4.6	5.68	5.6
Sample #7	80	116.8	4.89	4.2	5.77	4.9
Sample #8	80	116.2	2.98	2.6	6.43	5.5
Low Sample #1	80	10.5	1.39	13.3	1.84	17.6
Low Sample #2	80	12.7	1.14	9.0	2.04	16.0

Linearity/Reportable Range

To establish the linearity of the assay, study design was used based on the CLSI protocol EP6-A: Evaluation of the Linearity of Quantitative Measurement Procedures: a Statistical Approach: Approved Guideline.

Eleven levels of linearity were prepared by diluting a high serum sample containing 145.6 ng/mL r of 25-OH Vitamin D with a low serum sample containing 5.0 ng/mL of 25-OH Vitamin D, as measured by a the predicate device. The linearity levels were tested with the Diazyme 25-OH Vitamin D EIA, in triplicates. The results were processed using the EP Evaluator Software (Version 8.0) parameterized to an allowable systematic error of 7.8%. The assay was found to be linear between 6.7 ng/mL and 143.6 ng/mL. However, because the limit of quantitation of the assay was found to be 8.3 ng/mL, the claimed linearity range for the Diazyme 25-OH Vitamin D EIA is therefore 8.3 ng/mL to 143.6 ng/mL.

LoB/LoD/LoQ

The Limit of Blank (LoB), the Limit of Detection (LoD) and the Limit of Quantitation (LoQ) of the Diazyme 25-OH Vitamin D assay on microplate were determined according to CLSI EP17-A: Protocols for Determination of Limits of Detection and Limits of Quantitation. The following are the limits determined with the Diazyme 25-OH Vitamin D EIA:

LoB = 3.0 ng/mL

LoD = 5.6 ng/mL

LoQ = 8.3 ng/mL

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Analytical specificity

Interference Study

The Diazyme 25-OH Vitamin D EIA was subjected to an interference study according the CLSI EP7-A2 protocol. The following substances normally present in the blood produced less than 10% deviation when tested at levels equal to the concentrations listed below:

Interference	Concentration
Ascorbic Acid	10 mM
Free Bilirubin	40 mg/dL
Conjugated Bilirubin	40 mg/dL
Triglyceride	450 mg/dL
Hemoglobin	500 mg/dL

Cross Reactivity

Specificity of the Diazyme 25-OH Vitamin D EIA Assay was determined by adding Vitamin D metabolites to serum pool samples. Based on the results summarized in table below, the assay did not cross react with Vitamin d2 and Vitamin D3 and the assay recovers both 25-OH Vitamin D2 and 25-OH Vitamin D3 similarly. Although, the vitamin D metabolites 1, 25-OH and 24R, 25-OH showed cross reactivity as indicated below, these metabolites do not interfere with assay results when tested up to 100X the normal levels found in human sera.

Cross-reactant	Unspiked Vitamin D (ng/mL)	Spiked Vitamin D (ng/mL)	% Cross-reaction*
25-OH Vitamin D3(tested at 63 ng/ml)	24.4	87.4	100.0%
25-OH Vitamin D2(tested at 63 ng/ml)	24.4	85.4	96.8%
Vitamin D3(tested at 63 ng/ml)	24.4	24.2	-0.3%
Vitamin D2(tested at 63 ng/ml)	24.4	22.3	-3.3%
1;25-OH Vitamin D3(tested at 63 ng/ml)	24.4	84.1	94.8%
1,25-OH Vitamin D2(tested at 63 ng/ml)	24.4	62.0	59.7%
24R,25-OH Vitamin D3(tested at 56 ng/mL)	9.8	23.2	23.9%
3-epi-25-OH Vitamin D3(tested at 57 ng/ml)	21.2	69.6	84.9%
3-epi-25-OH Vitamin D2(tested at 57 ng/ml)	21.2	59.9	67.9%

*Cross-reactivity was calculated as (spiked sample value - unspiked sample value/concentration spiked) *100

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Comparison Studies

Method Comparison

Human serum samples were tested with the Diazyme 25-OH Vitamin D EIA and the obtained results were compared to the predicate method. A total of 58 serum samples were in this experiment. To ensure that the tested concentrations of 25-OH Vitamin D were distributed across the reportable dynamic range, 7 samples in the set were spiked with 25-OH Vitamin D stock solution of 25-OH Vitamin D3 and measured with the Diazyme and the IDS 25-OH Vitamin D assays. Using this study, we found that the Diazyme 25-OH Vitamin D EIA correlated with the predicate method with the following results:

	Serum Samples
n	58
Slope	0.941
Intercept	+1.448
Correlation coeffi-	0.930
Range of values	11.9 ng/mL- 131.5 ng/mL

Matrix Comparison

To evaluate the effect of anticoagulants, the Diazyme 25-OH Vitamin D EIA was used to measure the 25-OH Vitamin D concentrations of matched sets of serum, K3-EDTA plasma and Li-Heparin plasma. The reported values for each sample and for each matrix were obtained from single measurements. The total number of matched sets tested was 66. In order to cover the claimed measuring range for each matrix, seven spiked patient samples were included in the study.

Linear regression of the "Li-Heparin plasma versus Serum" data yielded the following results: y = 0.993 x + 1.855 and R2 = 0.961.

Linear regression of the "K3-EDTA plasma versus Serum" data yielded the following results: y = 1.006x + 2.901 and R2 = 0.967.

Reference Range Study

To determine a reference range for the Diazyme 25-OH Vitamin D EIA, the 25-OH Vitamin D serum concentrations of a US population of 157 apparently healthy individuals were measured with the Diazyme method. The individual patient serum samples used in this study were obtained from certified commercial sources (ProMedDx, LLC and Dx Biosamples). Forty seven (47) samples from Pennsylvania (Northern U.S.) were collected from an FDA Licensed Donor Center with informed consent. Fifty six (56) samples from Tennessee (Central U.S.) and Fifty four (54) samples from Texas (Southern U.S.) were collected according to an IRB approved protocol.

All participating individuals met the following inclusion conditions:

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The age of all individuals was within the 21-80 years old range.
Individuals were from three different geographical locations: 47 from Pennsylvania . (Northern US), 56 from Tennessee (Central US) and 54 from Texas (Southern US).
· All samples were collected during the months of October and November (fall season).
The studied population consisted of 72 light skin individuals (46%) and 85 dark skin in-. dividuals (54%).
155 individuals (98.7%) did not take any artificial Vitamin D supplements. Two individuals (1.3%) did take some Vitamin D supplements but did not exceed the daily dose of 2000 IU.
All 157 individuals did not have any family history of parathyroid or calcium regulatory . disease.
All 157 individuals did not have any history of kidney disease, GI disease, liver disease, . calcium-levels related disease, thyroid disease, parathyroid disease, calcium related disease, seizures, chronic disease or bariatric surgery.
All 157 individuals were not currently taking any medications that are known to affect . absorption or catabolism of Vitamin D (including cholesterol absorption inhibitors such as Vytorin®, Inegy™ or Zetia; anticonvulsants such as Neurontin, Depakine® and Trileptal; glucocorticoids such as Cortisol, Prednisone and Dexamethasone; HAART (AIDS treatment) or antirejection medications.

Analysis of the reference range study data yielded the following results:

Lowest 25-OH Vitamin D concentration: 8.4 ng/mL. .
· · · Highest 25-OH Vitamin D concentration: 61.3 ng/mL.
Median 25-OH Vitamin D concentration: 29.1 ng/mL
Observed range (2.5th to 97.5th percentile): 12.0 to 55.0 ng/mL.

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DEPARTMENT OF HEALTH & HUM AN SERVICES

Image /page/8/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo consists of a stylized depiction of an eagle or bird-like figure with three curved lines representing its wings or body. The logo is surrounded by text that reads "DEPARTMENT OF HEALTH & HUMAN SERVICES • USA" in a circular arrangement.

Public Health Service

Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-002

November 28, 2012

Diazyme Laboratories c/o Dr. Abhijit Datta 12889 Gregg Court Poway, CA 92064

Re: K122420

Trade/Device Name: Diazyme 25-OH Vitamin D EIA Kit Diazyme 25-OH Vitamin D EIA Control Kit Regulation Number: 21 CFR 862.1825 Regulation Name: Vitamin D Test System Regulatory Class: Class II Product Code: MRG, JJX Dated: October 16, 2012 Received: October 18, 2012

Dear Dr. Datta:

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.

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 Part 801); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set

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Page 2 - Dr. Abhijit Datta

forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

If you desire specific advice for your device on our labeling regulation (21 CFR Parts 801 and 809), please contact the Office of In Vitro Diagnostics and Radiological Health at (301) 796-5450. 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 Small Manufacturers, International and Consumer Assistance at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address http://www.fda.gov/cdrh/industry/support/index.html.

Sincerely yours,

Carol C. Benson 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): K122420

Device Name: Diazyme 25-OH Vitamin D EIA Kit and Diazyme 25-OH Vitamin D EIA Control Kit

Indications for Use:

The 25-OH Vitamin D EIA Control Kit is intended for use as quality controls for the Diazyme 25-OH Vitamin D EIA Kit only. For in vitro diagnostic use only.

Prescription Use _ X (21 CFR Part 801 Subpart D) And/Or

Over the Counter Use (21 CFR Part 801 Subpart C)

(PLEASE DO NOT WRITE BELOW THIS LINE; CONTINUE ON ANOTHER PAGE IF NEEDED)

Concurrence of CDRH, Office of In Vitro Diagnostics and Radiological Health (OIR)

Yung Chan

Division Sign-Off Office of In Vitro Diagnostics and Radiological Health

510(k) K122420

Regulation Number and Section

§ 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.