DIAZYME 25-HYDROXY VITAMIN D ASSAY KIT MODEL DZ688A, DIAZYME 25-HYDROXY VITAMIN D ASSAY CALIBRATOR SET MODEL DZ688A-CAL,

{0}------------------------------------------------ K10243.2 ## 510(k) SUMMARY JAN 1 4 2011 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 | |--------------------------------|----------------------------------------------------------------------------------------------------------------------------------| | Submitter's address: | 12889 Gregg Court<br>Poway, CA 92064<br>USA | | Name of Contact Person: | Dr. Abhijit Datta<br>Diazyme Laboratories<br>12889 Gregg Court<br>Poway, CA 92064<br>Phone: 858-455-4762<br>Fax: 858-455-2120 | | Date the Summary was Prepared: | Aug, 25, 2010; revised Jan 5, 2011 | | Name of the Device | 25-OH Vitamin D Assay Kit<br>25-OH Vitamin D Assay Calibrator Set<br>25-OH Vitamin D Assay Control Kit | | Trade Name: | 25-OH Vitamin D Assay Kit<br>25-OH Vitamin D Assay Calibrator Set<br>25-OH Vitamin D Assay Control Kit | | Common/Usual Name | Vitamin D Assay | | Device Classification Name | Vitamin D Test System | | Product code: | MRG – Vitamin D Test System<br>JIS - Calibrator, Primary<br>JJX – Single (specified) Analyte Controls (Assayed and<br>Unassayed) | | Panel: | Chemistry (75) | | Submission Type | 510k | {1}------------------------------------------------ | Regulation Number | 21 CFR 862.1825 – Vitamin D Test System<br>21 CFR 862.1150 - Calibrator, Primary<br>21 CFR 862.1660 - Quality Control material (Assayed and Unassayed) | |----------------------------|--------------------------------------------------------------------------------------------------------------------------------------------------------| | Device Class | II (Assay)<br>II (Calibrator)<br>I (Controls), | | Predicate Device: | The 25-OH Vitamin D Assay is substantially equivalent<br>to the currently marketed 25-OH Vitamin D EIA<br>(k021163). | | Manufacturing Address | Diazyme Laboratories<br>12889 Gregg Court<br>Poway, CA 92121<br>USA | | Establishment Registration | 2032900 | DESCRIPTION OF THE DEVICE ## DESCRIPTION OF THE DEVICE The 25-hydroxy (25-OH) Vitamin D assay is based on the principle of x-complementation of the enzyme B-galactosidase and the competition between an enzyme donor-25-OH Vitamin D conjugate, Vitamin D binding protein and the 25-OH Vitamin D content of a serum sample. Samples with higher 25-OH Vitamin D concentrations produce higher ß-galactosidase activities and vice versa. Chlorophenol red-B-D- galactopyranoside (CPRG) is used as the enzyme substrate and the accumulation of the reaction's product (chlorophenol red) is monitored at 560 nm. The 25-OH Vitamin D concentration of a patient sample is proportional to the measured ß-galactosidase activity. The assay consists of a sample extraction step during which Vitamin D is irreversibly dissociated from its serum transporter. Extracted samples are then analyzed on a microplate reader after the sequential addition of three reagents. 25-OH Vitamin D assay calibrator set is intended for use with the Diazyme 25-OH Vitamin D assay kit only. Six calibration levels are needed for each run. Calibrators are treated exactly the same as patient samples. 25-OH Vitamin D assay 2-point control set is intended for use with the Diazyme 25-OH Vitamin D assay 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 and run controls each time a new lot of reagents are received. {2}------------------------------------------------ ## INDICATIONS FOR USE The Diazyme 25-OH Vitamin D assay 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 assay calibrator set is intended for use in the calibration of the Diazyme 25-OH Vitamin D assay kit only. For in vitro diagnostic use only. The 25-OH Vitamin D assay control kit is intended for use as quality controls for the Diazyme 25-OH Vitamin D assay kit only. For in vitro diagnostic use only. | | Table 1 Summary of Assay Kit Components | | | | |--|-----------------------------------------|--|--|--| |--|-----------------------------------------|--|--|--| | IDS 25-OH Vitamin D EIA | Diazyme 25-OH Vitamin D Assay | |----------------------------------------------------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------| | (predicate k021163) | | | Kit can be only used for the 25-OH Vitamin D<br>quantification on microplate readers. | Kit can be only used for the 25-OH Vitamin D<br>quantification on microplate readers. | | Antibody Coated Plate (MICROPLAT) | Microplate | | Microplate with 25-OH Vitamin D sheep<br>polyclonal antibody linked to the inner<br>surface of the polystyrene wells, 12 x 8<br>well strips in a foil pouch with desicant. | Non-coated, polystyrene, round bottom,<br>microplate (96 wells, in a plastic pouch).<br>Microtube rack, 8-well microtube strips. | | Adhesive plate scaler | Adhesive plate sealer | | 8 per kit | 3 per kit | | Dissociation buffer (25-D Biotin)<br>1 bottle | Extraction solution (EX)<br>1 bottle | | Proprietary reagent for dissociating Vita-<br>min D 25-OH Vitamin D labeled with Biotin Stabilizers | Acetonitrile containing solution | | Wash Buffer (WASHBUF)<br>1 bottle | NA | | Phosphate buffered saline containing<br>Tween | | | Enzyme Conjugate (ENZYMCONJ)<br>1 bottle | Reagent 1<br>2 bottles | | Phosphate buffered saline containing<br>avidin linked to horseradish peroxidase,<br>protein, enzyme stabilizers and preserva-<br>tives. | R1a (1 bottle): Lyophilized enzyme<br>acceptor, Vitamin D binding protein<br>and phosphate salts. R1 (1 bottle): Reconstitution buffer<br>containing phosphate salts and pre-<br>servatives. | | TMB substrate (SUBS)<br>1 bottle | Reagent 2<br>2 bottles | {3}------------------------------------------------ | Proprietary aqueous formulation of<br>tetramethylbenzidine (TMB) and hydrogen<br>peroxide. | R2a (1 bottle): Lyophilized enzyme donor. R2 (1 bottle): Reconstitution buffer containing phosphate salts and preservatives. | |-------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------| | NA | Reagent 3<br>2 bottles R3a (1 bottle): Lyophilized substrate (CPRG). R3 (1 bottle): Reconstitution buffer containing phosphate salts and preservatives. | | STOP solution (HCl)<br>1 bottle Proprietary aqueous formulation of tetramethylbenzidine (TMB) and hydrogen peroxide. | STOP reagent<br>1 bottle Concentrated sodium carbonate solution. | | Calibrator set<br>1 x 1.0 mL Calibrator 0<br>1 x 1.0 mL Calibrator 1<br>1 x 1.0 mL Calibrator 2<br>1 x 1.0 mL Calibrator 3<br>1 x 1.0 mL Calibrator 4<br>1 x 1.0 mL Calibrator 5<br>1 x 1.0 mL Calibrator 6 | Calibrator set<br>1 x 1.0 mL Calibrator 1<br>1 x 1.0 mL Calibrator 2<br>1 x 1.0 mL Calibrator 3<br>1 x 1.0 mL Calibrator 4<br>1 x 1.0 mL Calibrator 5<br>1 x 1.0 mL Calibrator 6 | | Control set<br>1 x 1.0mL Control 1<br>1 x 1.0mL Control 2 | Control set<br>1 x 1.0mL Control 1<br>1 x 1.0mL Control 2 | # PERFORMANCE TESTING SUMMARIES ## Precision Study Acceptance Criteria: Precision: CV ≤ 10% The precision of the Diazyme 25-OH Vitamin D Microplate Assay is evaluated according to the Clinical and Laboratory Standards Institute (CLSI) EP5-A guideline. Acceptance Criteria: Precision: CV ≤ 10%. Seven precision levels were used: three serum controls (containing 21.5 ng/mL, 37.0 ng/mL and 60.8 ng/mL) and four serum samples correspond to deficient patients (19.0 ng/mL and 19.4 ng/mL), one sample corresponds to a near cut-off concentration (36.4 ng/mL) and one sample corresponds to an optimum level (43.4 ng/mL). {4}------------------------------------------------ Furthermore, to test the performance of the assay at the limits of the dynamic range, a severely deficient patient (7.8 ng/mL) and a very high sample (101.2 ng/mL) were subjected to the same precision studies. | | Deficient<br>Control | Normal<br>Control-1 | Normal<br>Control-2 | Deficient<br>Sample#1 | Deficient<br>Sample#2 | Near<br>cut-off<br>Sample | Optimal<br>Sample | |-----------------------------|----------------------|---------------------|---------------------|-----------------------|-----------------------|---------------------------|-------------------| | Data points | 40 | 40 | 40 | 40 | 40 | 40 | 40 | | Mean<br>(ng/mL) | 24.4 | 38.4 | 61.7 | 19.6 | 19.0 | 33.0 | 42.9 | | Within-run<br>SD<br>(ng/mL) | 1.0 | 2.0 | 4.0 | 0.9 | 1.1 | 1.5 | 1.6 | | Within-run<br>CV (%) | 4.2 | 5.3 | 6.5 | 4.8 | 5.7 | 4.6 | 3.6 | | Total SD<br>(ng/mL) | 1.9 | 3.7 | 5.9 | 1.8 | 1.7 | 2.8 | 3.7 | | Total CV<br>(%) | 7.9 | 9.7 | 9.6 | 9.4 | 9.1 | 8.4 | 8.6 | The mean value (Mean), standard deviation, within-run imprecision and total imprecision are calculated and summarized in the following tables: | | Severely Deficient Sample | Very High Sample | |-----------------------|---------------------------|------------------| | Data points | 40 | 40 | | Mean (ng/mL) | 7.8 | 101.2 | | Within-run SD (ng/mL) | 1.2 | 6.6 | | Within-run CV (%) | 15.3% | 6.5% | | Total SD (ng/mL) | 1.3 | 9.8 | | Total CV (%) | 16.7% | 9.7% | ## Linearity/Reportable Range To establish the linearity of the assay, study design was used 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 serum sample containing 130 ng/mL of 25-OH Vitamin D with a Vitamin D Diluent (Phosphate Buffer Saline supplemented with 9% of Bovine Serum Albumin). Linearity levels were prepared according to Clinical and Laboratory Standards Institute (CLSI) EP6-A. The samples prepared were tested with the Diazyme 25-OH Vitamin D assay, in triplicates. The results were processed using the EP Evaluator Software (Version 8.0) parameterized to an allowable systematic error of 10.8%. The linearity range was found to be 5.9 -120.2 ng/mL. {5}------------------------------------------------ ## 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 assay method: LoB = 1.0 ng/mL LoD = 3.6 ng/mL LoQ = 5.9 ng/mL ## Analytical specificity #### Interference Study The Diazyme 25-OH Vitamin D assay 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 | 30 mg/dL | | Triglyceride | 500 mg/dL | | Hemoglobin | 100 mg/dL | #### Cross Reactivity The Diazyme 25-OH Vitamin D assay was tested for cross reactivity to the following Vitamin D metabolites: | Cross reactant | Concentration<br>tested | Cross reactivity | |--------------------|-------------------------|------------------| | 25-OH Vitamin D3 | 100 ng/mL | 100.0% | | 25-OH Vitamin D2 | 100 ng/mL | 92.2% | | 1,25-OH Vitamin D3 | 100 ng/mL | 3.9% | | 1,25-OH Vitamin D3 | 100 ng/mL | 2.6% | | Vitamin D3 | 100 ng/mL | -0.6% | | Vitamin D2 | 100 ng/mL | 2.9% | {6}------------------------------------------------ ## Comparison Studies ## Method Comparison Human serum samples were tested with the Diazyme 25-OH Vitamin D assay and the obtained results were compared to the predicate method. A total of 80 samples (ranging from 7.7 to 113.8 ng/mL of 25-OH Vitamin D) were tested in both assays. The above described accuracy study showed that the Diazyme 25-OH Vitamin D Microplate Assay results correlated well with predicate method with a correlation coefficient of 0.936 with a slope of 1.039 and -2.469 intercept. | | Serum Samples | |---------------------|-----------------------| | n | 79 | | Slope | 1.039 | | Intercept | -2.481 | | Correlation coeffi- | 0.935 | | Range of values | 7.7 ng/mL-113.8 ng/mL | ## Matrix Comparison 26 samples were used in a comparison study between serum and Li-Heparin plasma. Linear regression of the "Serum versus Li-Heparin plasma" data yielded the following results: y = 0.992x-0.173 and R2 = 0.986. 55 samples were used in a comparison study between serum and K3-EDTA plasma. Linear regression of the "Serum versus K3-EDTA plasma" data yielded the following results: y = 0.908x + 1.749 and R2 = 0.981. ## Reference Range Study To determine a reference range for the Diazyme 25-OH Vitamin D microplate assay, the 25-OH Vitamin D serum concentrations of a US population of 150 apparently healthy individuals were measured with the Diazyme method. Thirty (30) samples from Pennsylvania (Northern U.S.) were collected from an FDA Licensed Donor Center with informed consent through Dx Biosamples. Sixty (60) samples from Tennessee (Central U.S.) and sixty (60) samples from Texas (Southern U.S.) were collected according to an IRB approved protocol by ProMedDx, LLC. All participating individuals met the following inclusion conditions: - The age of all individuals was within the 21-90 years old range. . - Individuals were from three different geographical locations: 30 from Pennsylvania . (Northern US), 60 from Tennessee (Central US) and 60 from Texas (Southern US). - All samples were collected during the months of October and November 2010 (fall sea-. son). - The studied population consisted of 63 light skin individuals (42%) and 87 dark skin in-. dividuals (58%). {7}------------------------------------------------ - 148 individuals (98.6%) did not take any artificial Vitamin D supplements. 2 individuals . (1.4%) did take some Vitamin D supplements but did not exceed the dose of 2000 IU. - All 150 individuals did not have any family history of parathyroid or calcium regulatory . disease. - . All 150 individuals did not have any history of kidney disease. Iiver disease, calcium-levels related disease, thyroid disease, parathyroid disease, calcium related disease, seizures, chronic disease or bariatric surgery. - All 150 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: 9.9 ng/mL. . - Highest 25-OH Vitamin D concentration: 65.8 ng/mL. . - . Median 25-OH Vitamin D concentration: 21.1 ng/mL - Observed range (2.5th to 97.5th percentile): 11.3 to 41.4 ng/mL. . {8}------------------------------------------------ #### DEPARTMENT OF HEALTH & HUMAN SERVICES Image /page/8/Picture/1 description: The image shows the seal of the U.S. Department of Health & Human Services. The seal is circular and contains the words "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" around the top half of the circle. In the center of the seal is a stylized image of an eagle. Food and Drug Administration 10903 New Hampshire Avenue Silver Spring, MD 20993 Diazyme Laboratories, Inc. c/o Abhijii Datta Quality Assurance Specialist 12889 Gregg Court Poway, CA 92064 JAN 1 4 201 Re: k102432 Trade Name: Diazyme 25-Hydroxy Vitamin D Microplate Assay Kit, Diazyme 25-Hydroxy Vitamin D Assay Calibrator Set, Diazyme 25-Hydroxy Vitamin D Assay Control Kit Regulation Number: 21 CFR §862.1825 Regulation Name: Vitamin D Test System Regulatory Class: Class II Product Codes: MRG, JIS, JJX Dated: January 5, 2011 Received: January 6, 2011 Dear Abhijit 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 rou mayy merce orovisions 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 such additional controls. Existing major regulations affecting your device it may or oubject to been from Regulations (CFR), Parts 800 to 895. 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 i least be advised that I DTC issuality over device complies with other requirements of the Act that IDA has made a decemmandions administered by other Federal agencies. You must Of any I-cuclar statures and regulations and admited to: registration and listing (21 comply with an the Free of requirements, and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); and good manufacturing practice mountal device related as vise enality systems (QS) regulation (21 CFR Part 820). {9}------------------------------------------------ Page 2 - 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 Diagnostic Device Evaluation and Safety 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-freenember (800) 638-2041 or (301) 796-7100 or at its Internet address http://www.fda.gov/cdrh/industry/support/index.html. Sincerely yours. CJC. Courtney Harper, Ph.D. Director Division of Chemistry and Toxicology Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health Enclosure {10}------------------------------------------------ # Indications for Use 510(k) Number k102432: Device Name: Diazyme 25-hydroxy Vitamin D Assay Kit Indications for Use: 机 机 The Diazyme 25-hydroxy Vitamin D Assay is designed for the quantification of total 25-hydroxy 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-hydroxy Vitamin D Assay Calibrator set is intended for use in the calibration of the Diazyme 25-OH Vitamin D Assay Kit only. For in vitro diagnostic use only. The 25-hydroxy Vitamin D Assay Control kit is intended for use as quality controls for the Diazyme 25-OH Vitamin D Assay kit only. For in vitro diagnostic use only. Prescription Use X (Part 21 CFR 801 Subpart D) AND/OR Over-The-Counter Use (21 CFR 801 Subpart C) (PLEASE DO NOT WRITE BELOW THIS LINE-CONTINUE ON ANOTHER PAGE IF NEEDED) Concurrence of CDRH, Office of In Vitro Diagnostic Devices (OIVD) Division Sign-Off Office of In Vitro Diagnostic Device Evaluation and Safety 510(k) K/02432 Page 1 of _ l Image /page/10/Picture/17 description: The image shows a black and white abstract pattern. The pattern consists of many small black dots scattered across a white background. The density of the dots varies, with the dots being more concentrated on the right side of the image and gradually becoming sparser towards the left. The overall effect is a gradient from a dense, dark area to a more open, light area.