1,25-DIHYDROXYVITAMIN D125I RIA

K014030 · DiaSorin, Inc. · MRG · Mar 26, 2002 · Clinical Chemistry

Device Facts

• Record ID: K014030
• Device Name: 1,25-DIHYDROXYVITAMIN D125I RIA
• Applicant: DiaSorin, Inc.
• Product Code: MRG · Clinical Chemistry
• Decision Date: Mar 26, 2002
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.1825
• Device Class: Class 2

Intended Use

The 1,25-Dihydroxyvitamin D 1251 RIA is a competitive equilibrium radioimmunoassay intended for the quantitative determination of 1,25-dihydroxyvitamin D (1,25-(OH)2-D) in human serum or EDTA plasma to be used to assess 1,25-(OH)2-D deficiency associated with renal disease. Since hydroxylation of circulating 25-hydroxyvitamin D to the biologically active form 1,25-(OH)2-D occurs in the kidney, renal disease may result in reduced levels of 1,25-(OH)2-D and compromised calcium metabolic homeostasis. Assay results should be used in conjunction with other clinical and laboratory data to assist the clinician in making individual patient management decisions in adult populations.

Device Story

Competitive equilibrium radioimmunoassay (RIA) for quantitative determination of 1,25-dihydroxyvitamin D in human serum or EDTA plasma. Procedure: sample extraction with acetonitrile to release metabolites from binding proteins and remove lipids; column chromatography on C18OH silica cartridges for purification. Treated samples incubated with rabbit anti-1,25-(OH)2 vitamin D antibody and 125I-radiolabeled tracer; bound/unbound separation via goat anti-rabbit polyethylene glycol precipitating complex. Centrifugation yields precipitate; radioactivity measured. Output: concentration in pg/mL derived from standard curve. Used in clinical laboratories by technicians. Results assist clinicians in managing patients with renal disease and compromised calcium homeostasis.

Clinical Evidence

Clinical trial conducted at three laboratories with 123 healthy donors and 87 ESRD patients. Established reference ranges: 19.8-68.0 pg/mL (normals) vs 1.6-17.3 pg/mL (ESRD). Precision evaluated via ANOVA across 16 assays over 110 days; total %CV ranged from 11.6% to 16.2% for low, mid, and high concentration samples. Fresh vs frozen sample equivalency confirmed via ANOVA (p-values 0.49 and 0.29).

Technological Characteristics

Competitive RIA; 125I-radiolabeled tracer; rabbit polyclonal antibody; goat anti-rabbit polyethylene glycol precipitating complex; acetonitrile extraction; C18OH silica cartridge chromatography; manual/laboratory-based; quantitative output via standard curve.

Indications for Use

Indicated for adult patients to assess 1,25-dihydroxyvitamin D deficiency associated with renal disease using human serum or EDTA plasma.

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

• DiaSorin 25-Hydroxyvitamin D 1251 RIA

Submission Summary (Full Text)

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K014030

#### APPENDIX E

As required by 21 CFR 807.3, a summary of the 510(k) safety and effectiveness information contained in this submission is provided as Appendix E.

## 510(k) SUMMARY

SUBMITTED BY:

Diana Clive DiaSorin, Inc. 1951 Northwestern Ave. P.O. Box 285 Stillwater, MN 55082-0285 [phone] voice [phone] fax November 30, 2001

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.

NAME OF DEVICE: Trade Name:

1,25-Dihydroxyvitamin D 1251 RIA

Common Names/Descriptions:

the the Immunoassay for quantitative determination of 1,25 dihydroxyvitamin D (1,25-(OH)2-D)

Classification Name:

Vitamin D test system

PREDICATE DEVICE:

DiaSorin 25-Hydroxyvitamin D 1251 RIA

The 1,25-Dihydroxyvitamin D 1251 RIA assay is a competitive INTENDED USE: equilibrium radioimmunoassay intended for the quantitative determination of 1,25 dihydroxyvitamin D (1,25-(OH)2-D) in human serum or EDTA plasma to be used to assess 1,25-(OH)2-D deficiency associated with renal disease. Assay results should be used in conjunction with other clinical and laboratory data to assist the clinician in making individual patient management decisions in adult populations.

DEVICE DESCRIPTION: The 1,25-Dihydroxyvitamin D 1251 RIA is a competitive radioimmunoassay intended for the quantitative determination of 1,25 Dihydroxyvitamin D (1,25-(OH)2-D) in human serum or EDTA plasma. The assay consists of a two-step procedure. Serum or plasma patient samples as well as standards and kit controls are first extracted with acetonitrile to free the 1,25-(OH)2 vitamin D2 and D3 from their vitamin D binding protein, and remove lipids that might interfere with the assay. The metabolites are then extracted by column chromatography on C18OH silica cartridges using a series of organic solvent washes. Following the extraction, the treated samples are assayed using a

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competitive radioimmunassay procedure. The primary antibody (rabbit anti-1,25-(OH)2 vitamin D) is highly specific for both 1,25(OH)2 vitamin D3 and 1,25(OH)2.vitamin D2. Vitamin D) is inghty specific for both 1,25(011)2 (011)2 vitamin D tracer Daring the binding sites on the primary antibody. Separation of bound and unbound vitamin D2 or D3 is accomplished using a goat anti-rabbit (GAR) polyethylene glycol precipitating complex. After an incubation and centrifugation, sample precipitates are proofitaing complex. The amount of radioactivity in the precipitate is inversely proportional to the concentration of 1,25-(OH)2 vitamin D in the sample. Values are proportional to the coma standard curve of known calibrators and expressed as pg/ml.

| Feature | 25-OH-D | 1,25-(OH)2-D |
|----------------------------|---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|
| Intended Use | FOR IN VITRO DIAGNOSTIC USE.<br>This kit is intended for the quantitative<br>determination of 25-hydroxyvitamin D<br>(25-OH-D) and other hydroxylated<br>vitamin D metabolites in human serum<br>or plasma to be used in the assessment<br>of vitamin D sufficiency. Assay<br>results should be used in conjunction<br>with other clinical and laboratory data<br>to assist the clinician in making<br>individual patient management<br>decisions in an adult population. | FOR IN VITRO DIAGNOSTIC USE.<br>This kit is intended for the quantitative<br>determination of 1,25-dihydroxy-<br>vitamin D (1,25-(OH)2-D) in human<br>serum or EDTA plasma to be used to<br>assess 1,25-(OH)2-D deficiency<br>associated with renal disease. Assay<br>results should be used in conjunction<br>with other clinical and laboratory data<br>to assist the clinician in making<br>individual patient management<br>decisions in adult populations. |
| Assay Type | RIA | RIA |
| Acetonitrile<br>Extraction | For isolation of 25-(OH)-D | For isolation of 1,25-(OH)2-D |
| Antiserum | Polyclonal specific for 25-(OH)-D2/D3 | Polyclonal specific for 1,25-(OH)2-D2/D3 |
| Tracer | 125I radiolabeled 25-(OH)-D analog | 125I radiolabeled 1,25-(OH)2-D3 analog |
| Precipitating<br>Complex | 20 minute incubation at 20-25° C with<br>second antibody precipitating complex | 20 minute incubation at 20-25° C with<br>second antibody precipitating complex |
| Kit Controls | Two levels, human serum based,<br>extracted identical to standards and<br>patient samples. | Two levels, human serum based,<br>extracted identical to standards and<br>patient samples. |
| Standards | Five levels, human serum based,<br>extracted identical to controls and<br>patient samples. | Five levels, human serum based,<br>extracted identical to controls and<br>patient samples. |

# TECHNOLOGICAL COMPARISON TO PREDICATE:

PERFORMANCE DATA: A clinical trial was conducted at three independent clinical laboratories. Serum values for 1,25(OH)2-vitamin D were collected from two distinct populations: apparently healthy normal donors, and patients with end-stage renal disease (ESRD). Reference ranges established in the trial are summarized below.

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| All Sites | 1,25(OH)2D (pg/mL) | | | |
|------------|--------------------|------|------|-----------|
| Population | n | Mean | 2 SD | Range |
| Normals | 123 | 43.9 | 24.1 | 19.8-68.0 |
| ESRD's | 87 | | | 1.6-17.3 |

The equivalency of assay samples as either fresh serum or serum that has been frozen and The equivalely of about bangles mory table with 95% ANOVA determined P-values is presented below.

| | Normals | | ESRD's | |
|-------|---------|-------|--------|-------|
| | Frozen | Fresh | Frozen | Fresh |
| N | 72 | 51 | 70 | 17 |
| Mean | 43.2 | 44.8 | 6.1 | 5.3 |
| ANOVA | 0.49 | | 0.29 | |

Reproducibility was established both at DiaSorin and during the clinical trial. Three human serum-based samples with 1,25-(OH)2-D concentrations distributed across the numan berain based calliples with 5 assay days at DiaSorin, spanning more than 60 Multiple technicians, as well as multiple lot numbers for all operating days. operaing were included. The combined results were evaluated by analysis of variance (ANOVA) and are summarized in the following table.

| | | | Within-run | | Between Day | | Total | |
|--------|----|-----------------|------------|-------|-------------|-------|-------|-------|
| Sample | n | Mean<br>(pg/mL) | S.D. | %C.V. | S.D. | %C.V. | S.D. | %C.V. |
| Low | 25 | 25.8 | 1.76 | 6.8 | 3.8 | 14.6 | 4.0 | 15.3 |
| Mid | 25 | 41.3 | 3.16 | 7.7 | 4.6 | 11.1 | 5.1 | 12.3 |
| High | 25 | 105.2 | 11.86 | 11.3 | 11.8 | 11.2 | 14.4 | 13.7 |

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These identical human serum-based controls were assayed during a clinical trial conducted at three different laboratories, over 16 different assays, spanning more than 110 days. The combined results from all three sites are summarized below.

| Sample | n | Mean<br>(pg/mL) | S.D. | %CV |
|--------|----|-----------------|------|------|
| Low | 16 | 24.2 | 3.9 | 16.2 |
| Mid | 16 | 41.0 | 5.8 | 14.1 |
| High | 16 | 97.7 | 11.3 | 11.6 |

Similarly, kit controls included with each kit were included in each of the 16 assays The following table above, as prescribed in the package insert instructions. summarizes the precision of the controls over the course of the clinical trial.

| | | Site A | Site B | Site C | Total |
|---------------|------|--------|--------|--------|-------|
| Kit Control 1 | n | 7 | 4 | 8 | 19 |
| | Mean | 24.7 | 27.2 | 25.5 | 25.6 |
| | SD | 3.1 | 3.3 | 4.0 | 3.5 |
| | %CV | 12.5 | 12.2 | 15.8 | 13.7 |
| Kit Control 2 | n | 7 | 4 | 7 | 18 |
| | Mean | 66.2 | 59.9 | 67.2 | 65.2 |
| | SD | 5.1 | 4.2 | 11.1 | 8.1 |
| | %CV | 7.8 | 7.1 | 16.6 | 12.4 |

The data collected during the clinical trial described above fully CONCLUSION: substantiates the intended use of the 1,25-Dihydroxyvitamin D 1251 RIA. Reference ranges established for both a healthy population and one with end stage renal disease are fully distinct. This indicates that the DiaSorin 1,25-Dihydroxvitamin D 131 RIA can effectively discriminate between the two populations based on 1,25-(OH)2-D serum values. Assay results can be used to assess 1,25-(OH)2-D deficiency associated with renal disease.

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Image /page/4/Picture/2 description: The image shows the logo for the Department of Health & Human Services. The logo consists of a circular border with the text "DEPARTMENT OF HEALTH & HUMAN SERVICES USA" arranged around the perimeter. Inside the circle is a stylized symbol featuring three abstract shapes that resemble a person or a bird in flight. The shapes are layered on top of each other, creating a sense of movement and depth.

# MAR 2 6 2002

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

Mr. David M. Ikeda Manager, Regulatory Affairs and Quality Systems DiaSorin Inc. 1990 Industrial Blvd. P.O. Box 285 Stillwater, MN 55082-0285

Re: k014030

Trade/Device Name: 1,25-Dihydroxyvitamin D 1251 RIA Kit Regulation Number: 21 CFR 862.1825 Regulation Name: Vitamin D test system Regulatory Class: Class II Product Code: MRG Dated: February 18, 2002 Received: February 20, 2002

#### Dear Mr. Ikeda:

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 such 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); good manufacturing practice requirements as set 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.

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This letter will allow you to begin marketing your device as described in your 510(k) premarket notification. The FDA finding of substantial equivalence of your device to a legally marketed predicate device results in a classification for your device and thus, permits your device to proceed to the market.

If you desire specific advice for your device on our labeling regulation (21 CFR Part 801 and 1 additionally 809.10 for in vitro diagnostic devices), please contact the Office of Compliance at [phone]. Additionally, for questions on the promotion and advertising of your device, please contact the Office of Compliance at [phone]. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR 807.97). Other general information on your responsibilities under the Act may be obtained from the Division of Small Manufacturers International and Consumer Assistance at its toll-free number [phone] or [phone] or at its internet address "http://www.fda.gov/cdrh/dsma/dsmamain.html".

Sincerely yours,

Steven Butman

Steven I. Gutman, M.D., M.B.A. Director Division of Clinical Laboratory-Devices Office of Device Evaluation Center for Devices and Radiological Health

Enclosure

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## INDICATIONS FOR USE

510(k) Number (if known):

Device Name:

1,25-Dihydroxyvitamin D 1251 RIA Kit

K014030

Indications For Use:

The 1,25-Dihydroxyvitamin D 1251 RIA is a competitive equilibrium radioimmunoassay intended for the quantitative determination of 1,25-dihydroxyvitamin D (1,25-(OH)2-D) in human serum or EDTA plasma to be used to assess 1,25-(OH)2-D deficiency associated with renal disease. Since hydroxylation of circulating 25-hydroxyvitamin D to the biologically active form 1,25-(OH)2-D occurs in the kidney, renal disease may result in reduced levels of 1,25-(OH)2-D and compromised calcium metabolic homeostasis. Assay results should be used in conjunction with other clinical and laboratory data to assist the clinician in making individual patient management decisions in adult populations.

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

Concurrence of CDRH, Office of Device Evaluation (ODE)

Division Sign-Off

Carol C. Benson for Jean Cooper

vision of Clinical Laboratory D 510fki Number Prescription Use: the-Counter Use: (Per 21 CFR 801.109)