IMMULITE 2000 Vitamin B12 is for in vitro diagnostic use with the IMMULITE 2000 Analyzer for the quantitative measurement of vitamin B12 in serum or heparinized plasma, as an aid in clinical diagnosis and treatment of anemia.

Device Description

IMMULITE® 2000 Vitamin B12 is a clinical device for use with the IMMULITE 2000 Automated Immunoassay Analyzer. IMMULITE® and IMMULITE® 2000 Vitamin B12 are chemiluminescent immunoassays. The technology in DPC's IMMULITE® 2000 Vitamin B12 is a unique combination of technologies employed in previously cleared and commercially marketed DPC products. The IMMULITE 2000 Vitamin B12 assay begins with a one-cycle sample treatment of patient serum or plasma with dithiothreitol (DTT) and a sodium hydroxide/potassium cyanide solution (NaOH/KCN) in a reaction tube containing no bead. After a 30-minute incubation, the treated sample is transferred to a second reaction tube containing a vitamin B12-coated polystyrene bead, hog intrinsic factor (HIF) and an alkaline phosphatase-labeled antibody specific for HIF. During a 30-minute incubation, the vitamin B12 released from endogenous binding proteins during sample treatment competes with immobilized vitamin B12 for binding with HIF. Alkaline phosphatase-labeled anti-HIF antibody binds to HIF and is immobilized only if HIF is bound to the B12-coated bead. Unbound enzyme conjugate is removed by centrifugal wash. Substrate is added. The chemiluminescent substrate, a phosphate ester of adamantyl dioxetane, undergoes hydrolysis in the presence of alkaline phosphatase to yield an unstable intermediate. The continuous production of this intermediate results in the sustained emission of light, thus improving precision by providing a window for multiple readings. The bound comblex - and thus also the photon output, as measured by the luminometer - is inversely proportional to the concentration of vitamin B12 in the sample.

AI/ML Overview

Here's an analysis of the provided text, outlining the acceptance criteria and study details for the IMMULITE® 2000 Vitamin B12 device:

1. Table of Acceptance Criteria and Reported Device Performance

Acceptance Criteria	Reported Device Performance
Performance Equivalence - Method Comparison (Correlation with predicate device)	The IMMULITE® 2000 Vitamin B12 procedure was compared to DPC's IMMULITE® Vitamin B12 (predicate device). Linear regression yielded: (IMMULITE® 2000) = 0.95 (IMMULITE®) - 34 pg/mL. The correlation coefficient (r) was 0.937.
Mean values (Comparison of means with predicate device)	IMMULITE® 2000 Mean: 400 pg/mL. IMMULITE® Mean (Predicate): 457 pg/mL.

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

Sample Size: 166 patient serum samples for the method comparison study.
Data Provenance: The document does not specify the country of origin. It does not explicitly state if the data was retrospective or prospective, but the phrasing "patient serum samples" suggests existing samples were used, which would typically be retrospective.

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

This section is not applicable to this type of device. The ground truth for this device is established by a quantitative measurement using a legally marketed predicate device (DPC's IMMULITE® Vitamin B12), not by expert consensus or interpretation of images/data. The predicate device itself acts as the reference standard.

4. Adjudication Method for the Test Set

This section is not applicable. Since the comparison is against an established quantitative assay, there is no need for an adjudication method by experts to resolve disagreements, as would be the case in subjective diagnostic evaluation.

5. If a Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study was Done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance

This section is not applicable. The device is an in-vitro diagnostic (IVD) assay for quantitative measurement of a biomarker (Vitamin B12), not an AI-assisted diagnostic imaging or interpretation tool that would involve human readers.

6. If a Standalone (i.e., algorithm only without human-in-the-loop performance) was done

Yes, this was a standalone performance evaluation. The IMMULITE® 2000 Vitamin B12 device's performance was compared directly to the predicate IMMULITE® Vitamin B12 device without human intervention influencing the measurement results of either system. The method comparison described is a "standalone" evaluation of the new device's quantitative output against the established device's quantitative output.

7. The Type of Ground Truth Used

The ground truth for the comparison was the quantitative measurement of vitamin B12 in serum samples obtained using the IMMULITE® Vitamin B12 (the predicate device).

8. The Sample Size for the Training Set

The document does not explicitly mention a separate "training set". For this type of IVD, method comparison studies do not typically involve a training/test set split in the same way machine learning models do. The 166 patient samples were used for the performance comparison. If any internal validation or calibration was performed during development, that information is not provided here.

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

Since a training set is not explicitly mentioned as a distinct component of the regulatory submission (as would be for an AI/ML device), the method for establishing its ground truth is also not described. For an IVD, the development and calibration typically rely on reference materials, calibrators, and internal validation studies, rather than a "ground truth" derived from human experts for a training set. The performance is then validated against a predicate device.

Summary

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NOV 1 7 1999

K993251

510(k) Summary of Safety and Effectiveness

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

Name: Diagnostic Products Corporation Address: 5700 West 96th Street Los Angeles, California 90045-5597 (310) 645-8200 Telephone Number: (310) 645-9999 Facsimile Number: Contact Person: Edward M. Levine, Ph.D. Director of Clinical Affairs Date of Preparation: September 24, 1999 IMMULITE® 2000 Vitamin B12 Device Name: Reagent system for the determination of vitamin Trade: B12 in serum and heparinized plasma Catalog Number: L2KVB2 (200 tests); L2KVB6 (600 tests) Class II device, 75-CDD (21CFR 862.1810) Classification: Manufacturer: Diagnostic Products Corporation 5700 West 96th Street Los Angeles, California 90045-5597 DPC's Registration # is 2017183 Establishment Registration #: DPC's IMMULITE® Vitamin B12 Substantially Equivalent Predicate Device: IMMULITE® 2000 Vitamin B12 is a clinical device Description of Device: for use with the IMMULITE 2000 Automated Immunoassay Analyzer.

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Intended Use of the Device:

IMMULITE 2000 Vitamin B12 is for in vitro diagnostic use with the IMMULITE 2000 Analyzer - for the quantitative measurement of vitamin B12 in serum or heparinized plasma, as an aid in clinical diagnosis and treatment of anemia.

Summary and Explanation of the Test:

Vitamin B12 (cobalamin) and folate are nutrients essential to hematopoiesis. Megaloblastic anemia is almost always due to lack of one of these two vitamin B12 deficiency can also result in severe neurological impairment.

Circulating levels of vitamin B12 are usually a good index to tissue stores. That is, vitamin B12 levels as measured in serum or plasma by an optimized assay system are typically low in vitamin B12 deficiency, and normal or elevated otherwise. Exceptions to this rule can occur in those relatively uncommon situations where levels of vitamin B12 transport proteins are abnormal. Thus, low circulating vitamin B12 levels can occur in the absence of vitamin B12 deficiency where the level of transcobalamin I (a physiologically inactive transport protein) is low.

Conversely, vitamin B12 deficiency can occur in the presence of normal or even elevated plasma vitamin B12 levels where transcobalamin II levels are low or where levels of inactive vitamin B 12 transport proteins are high, as in chronic myelogenous leukemia. (Circulating folate levels are usually normal or elevated in vitamin B12 deficiency, but red cell folate levels are frequently low in this condition.)

Vitamin B12 deficiency occurs only rarely as a result of dietary lack of this vitamin. More commonly, it results from impaired absorption, as in partial or total gastrectomy, or in pernicious anemia, a condition characterized by absence or near absence of intrinsic factor. Since roughly two thirds of all patients with pernicious anemia have blocking antibodies to intrinsic factor (IFbAb), while IFbAb are only very rarely encountered in other situations, IFbAb determinations represent a useful follow-up test for the differential diagnosis of vitamin B12 deficiency. (Circulating intrinsic factor antibodies are present in more than half of all pernicious anemia patients. Increased transport protein levels can occur, for example in chronic myelogenous leukemia.)

Common causes of high vitamin B12 levels include liver disease, myeloproliferative disease (with chronic myelogenous leukemia as a special case) and the use of multivitamin supplements.

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Performance Equivalence - Technology Comparison:

IMMULITE® and IMMULITE® 2000 Vitamin B12 are chemiluminescent immunoassays. The technology in DPC's IMMULITE® 2000 Vitamin B12 is a unique combination of technologies employed in previously cleared and commercially marketed DPC products.

The IMMULITE 2000 Vitamin B12 assay begins with a one-cycle sample treatment of patient serum or plasma with dithiothreitol (DTT) and a sodium hydroxide/potassium cyanide solution (NaOH/KCN) in a reaction tube containing no bead. After a 30-minute incubation, the treated sample is transferred to a second reaction tube containing a vitamin B12-coated polystyrene bead, hog intrinsic factor (HIF) and an alkaline phosphatase-labeled antibody specific for HIF. During a 30-minute incubation, the vitamin B12 released from endogenous binding proteins during sample treatment competes with immobilized vitamin B12 for binding with HIF. Alkaline phosphatase-labeled anti-HIF antibody binds to HIF and is immobilized only if HIF is bound to the B12-coated bead. Unbound enzyme conjugate is removed by centrifugal wash. Substrate is added.

The chemiluminescent substrate, a phosphate ester of adamantyl dioxetane, undergoes hydrolysis in the presence of alkaline phosphatase to yield an unstable intermediate. The continuous production of this intermediate results in the sustained emission of light, thus improving precision by providing a window for multiple readings. The bound comblex - and thus also the photon output, as measured by the luminometer - is inversely proportional to the concentration of vitamin B12 in the sample.

IMMULITE® Vitamin B12 is a chemiluminescent version of the classic method for vitamin B12 radioassay, involving a preliminary heat denaturation step. Vitamin B12 in the patient sample is released from carrier proteins by incubation at 100 ℃ in the presence of dithiothreitol and potassium cyanide to inactivate vitamin B12-binding proteins, even at extreme levels, as well as antibodies to intrinsic factor.

After the heat denaturation step, the treated patient sample and purified hog intrinsic factor are simultaneously introduced into an IMMULITE® Test Unit containing a polystyrene bead coated with a B12 analog, and incubated for approximately 30 minutes at 37 ℃ with intermittent agitation. During this incubation, vitamin B12 in the treated sample competes with the B12 analog on the solid phase for a limited number of vitamin B12 binding sites on the purified intrinsic factor. (Endogenous vitamin B12 analogs do not interfere, because the binder is free of R-protein.) Alkaline phosphatase-labeled anti-hog intrinsic factor is introduced, and the Test Unit is incubated for another 30-minute cycle. The unbound enzyme conjugate is removed by a centrifugal wash.

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Performance Equivalence - Technology Comparison (continued):

Substrate is then added, and the Test Unit is incubated for a further 10 minutes. The chemiluminescent substrate, a phosphate ester of adamantyl dioxetane, undergoes hydrolysis in the presence of alkaline phosphatase to yield an unstable intermediate. The continuous production of this intermediate results in the sustained emission of light, thus improving precision by providing a window for multiple readings. The bound complex - and thus also the photon output, as measured by the luminometer - is inversely proportional to the concentration of vitamin B12 in the sample.

Performance Equivalence - Method Comparison:

The IMMULITE® 2000 Vitamin B12 procedure was compared to DPC's IMMULITE® Vitamin B12 on 166 patient serum samples. Linear regression yielded the following results.

(IMMULITE® 2000) = 0.95 (IMMULITE®) - 34 pg/mL

r = 0.937

400 pg/mL (IMMULITE® 2000) Means: 457 pg/mL (IMMULITE®)

Conclusion:

The data presented in this summary of safety and effectiveness is the data the Food and Drug Administration used in granting DPC substantial equivalence for IMMULITE® 2000 Vitamin B12.

edward Lewis

Edward M. Levine, Ph.D. Director of Clinical Affairs

9/24/99
Date

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Image /page/4/Picture/1 description: The image shows the logo for the Department of Health & Human Services USA. The logo is a circle with the words "DEPARTMENT OF HEALTH & HUMAN SERVICES · USA" around the perimeter. Inside the circle is a stylized image of three human profiles facing to the right, with three curved lines above them. The logo is black and white.

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

NOV 17 1999

Edward M. Levine, Ph.D. Director of Clinical Affairs Diagnostic Products Corporation 5700 West 96th Street Los Angeles, California 90045-5597

Re: K993251

Trade Name: IMMULITE® 2000 Vitamin B12 Regulatory Class: II Product Code: LIG Dated: September 24, 1999 Received: September 28, 1999

Dear Dr. Levine:

We have reviewed your Section 510(k) notification of intent to market the device referenced above and we 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). 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 (Premarket Approval), 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 895. A substantially equivalent determination assumes compliance with the Current Good Manufacturing Practice requirements, as set forth in the Quality System Regulation (QS) for Medical Devices: General regulation (21 CFR Part 820) and that, through periodic OS inspections, the Food and Drug Administration (FDA) will verify such assumptions. Failure to comply with the GMP regulation may result in regulatory action. In addition, FDA may publish further announcements concerning your device in the Federal Register. Please note: this response to your premarket notification submission does not affect any obligation you might have under sections 531 through 542 of the Act for devices under the Electronic Product Radiation Control provisions, or other Federal laws or regulations.

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Under the Clinical Laboratory Improvement Amendments of 1988 (CLIA-88), this device may require a CLIA complexity categorization. To determine if it does, you should contact the Centers for Disease Control and Prevention (CDC) at (770) 488-7655.

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 additionally 809.10 for in vitro diagnostic devices), please contact the Office of Compliance at (301) 594-4588. Additionally, for questions on the promotion and advertising of your device, please contact the Office of Compliance at (301) 594-4639. Also, please note the regulation entitled, "Misbranding by reference to premarket notification"(21 CFR 807.97). Other general information on your responsibilities under the Act may be obtained from the Division of Small Manufacturers Assistance at its toll-free number (800) 638-2041 or (301) 443-6597, or at its internet address "http://www.fda.gov/cdrh/dsma/dsmamain.html".

Sincerely yours,

Steven Sutman

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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510(k) Number (if known): Device Name: IMMULITE® 2000 Vitamin B12

Indications For Use:

Y au Cooper
(Division Sign-Off)

sion of Clinical Laboratory Devices ULI 510(k) Number

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

Concurrence of CDRH, Office of Device Evaluation (ODE)

✓

Prescription Use
(Per 21 CFR 801.109)

Over-The-Counter Use

(Optional Format 1-2-96)

Regulation Number and Section

§ 862.1810 Vitamin B

12 test system.(a)
Identification. A vitamin B12 test system is a device intended to measure vitamin B12 in serum, plasma, and urine. Measurements obtained by this device are used in the diagnosis and treatment of anemias of gastrointestinal malabsorption.(b)
Classification. Class II.