The Axis-Shield HoloTC RIA is an in-vitro diagnostic assay for quantitative measurement of holotranscobalamin (vitamin B12 bound to transcobalamin) in human serum or plasma. Measurements obtained by this device are used in the diagnosis and treatment of vitamin B12 deficiency. HoloTC RIA is calibrated with HoloTC Calibrators. HoloTC controls are assayed for the verification of the accuracy and precision of the HoloTC RIA.

Device Description

The Axis-Shield HoloTC RIA is a competitive binding immunoassay in which a specific monoclonal antibody is used to capture transcobalamin from the patient sample. Thereafter the procedure is as commonly used in vitamin B12 assays. The cobalamin (vitamin B12) is released from the transcobalamin using dithiothreitol and sodium hydroxide. The released cobalamin (vitamin B12) then competes for a limited amount of intrinsic factor with added 'Co labelled vitamin B12. The Axis-Shield HoloTC RIA differs from the predicate device in two main aspects:

1. The use of a transcobalamin specific antibody, this allows the quantitation of only the cobalamin bound to the protein transcobalamin as opposed to measurement of cobalamin bound to all proteins in the predicate device and
1. The detection signal is radioactivity (37Co) as opposed to chemiluminescence in the predicate device.

AI/ML Overview

Here's an analysis of the provided text regarding the Axis-Shield HoloTC RIA device, addressing your requested information:

1. Table of Acceptance Criteria and Reported Device Performance

The document does not explicitly state pre-defined acceptance criteria for all performance metrics. However, where implicitly stated or inferable, they are included.

Performance Characteristic	Acceptance Criteria (Implicit/Explicit)	Reported Device Performance
Method Comparison	Close linear correlation, ideally slope ≈ 1, intercept ≈ 0, high r²	HoloTC pmol/L = 0.55 * VB12-2 pmol/L; r² = 0.52 (compared to predicate Bayer Advia Centaur VB12 assay)
Reference Interval	n/a (established for diagnosis)	37-171 pmol/L (95% central interval for Finnish population, n=303)
External Evaluation (CV)	<15% (for three levels of serum samples)	CVs for three serum samples were "within the given acceptance criteria, <15 %"
Precision (Within-run CV)	Not explicitly stated but implied to be acceptable	11% (low), 5% (medium), 8% (high)
Precision (Total CV)	Not explicitly stated but implied to be acceptable	11% (low), 6% (medium), 8% (high)
Linearity (Working Range)	Meet all criteria for linearity	Demonstrated within 10-160 pmol/L
Linearity (Recovery Test)	Within acceptance criteria	Demonstrated within acceptance criteria
Linearity (Dilution Correlation)	High r², slope ≈ 1, intercept ≈ 0	r² = 0.998, slope = 1.01 ± 0.02, y-intercept = - 5 ± 2 pmol/L (for dilutions 13-182 pmol/L)
LOQ	Not explicitly stated/implied to be acceptable for clinical use	8-176 pmol/L
LOD	Not explicitly stated/implied to be acceptable for clinical use	6 pmol/L
Cross-reactivity (HC)	±10% (for up to 70,000 pg/mL HC)	Well within acceptance criteria
Interference	<10% interference	<10% for bilirubin, hemoglobin, total protein, lipids
Serum/EDTA Plasma Comparison	No significant difference	No significant difference demonstrated
In-use Stability	Not explicitly stated/implied to be acceptable	Stability up to three months after opening
Concordance	Not explicitly stated/implied to be acceptable for diagnosis	80% (weighted combined estimate)
Sensitivity	Not explicitly stated/implied to be acceptable for diagnosis	99.5% (weighted combined estimate)
Specificity	Not explicitly stated/implied to be acceptable for diagnosis	76.3% (weighted combined estimate)

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

Method Comparison: 392 patient samples. Data provenance not explicitly stated (e.g., country of origin, retrospective/prospective).
Reference Interval: 303 individuals from a "Finnish population of normal individuals." This suggests prospective collection for establishing normative data within that specific population.
External Evaluation: Three different levels of native and pooled serum samples, in addition to kit controls Low and High. Number of samples not given, but tested at two external study sites (UK and Denmark).
Precision Studies: Three levels of serum samples (low, medium, high). Number of samples for each level not explicitly stated, but duplicate measurements were performed.
Linearity (Dilution): High HoloTC sample diluted into six dilutions. The original sample size is not stated.
Serum/EDTA Plasma Comparison: 50 paired serum and EDTA plasma samples from the same donors.
Sensitivity and Specificity: Samples from 3 study sites: 112 classified as "likely vitamin B12 deficient" and 313 as "not likely vitamin B12 deficient." Total of 425 samples. Data provenance (country of origin, retrospective/prospective) not explicitly stated.

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

The document does not mention the use of experts to establish ground truth for the test set.
For sensitivity and specificity, samples were classified based on "cobalamin and MMA levels," indicating an objective biochemical measure rather than expert interpretation of a diagnostic image or clinical presentation.

4. Adjudication Method for the Test Set

No adjudication method is mentioned, as there is no indication of expert interpretation requiring consensus. The ground truth for classification of B12 deficiency was based on biochemical markers (cobalamin and MMA levels).

5. If a multi-reader multi-case (MRMC) comparative effectiveness study was done

No, an MRMC comparative effectiveness study was not done. This device is an in-vitro diagnostic assay which provides a quantitative measurement, not an imaging device requiring human interpretation, therefore, the concept of a multi-reader study or human readers improving with AI assistance is not applicable. The comparison was against a predicate assay (Bayer Advia Centaur VB12 assay).

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

Yes, the entire study focuses on the standalone performance of the Axis-Shield HoloTC RIA assay (which can be considered the "algorithm only" in the context of an IVD) in measuring holotranscobalamin levels. As an in-vitro diagnostic quantitative assay, its performance is inherently standalone, providing a numerical result.

7. The type of ground truth used

Method Comparison: The ground truth for comparison was the measurements obtained from the predicate device, the Bayer Advia Centaur VB12 assay.
Reference Interval: The ground truth was based on the measured HoloTC levels in a population of "normal individuals."
Sensitivity and Specificity: The ground truth for classifying samples as "likely vitamin B12 deficient" or "not likely vitamin B12 deficient" was based on cobalamin and MMA levels. This is a form of outcomes data or established biochemical markers for the condition.

8. The Sample Size for the Training Set

The document does not explicitly mention a "training set" in the context of an AI/ML model development. For an immunoassay like the HoloTC RIA, the "training" involves the development and optimization of the assay reagents, protocols, and calibration curves. The provided numbers are for validation or evaluation sets.

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

As there is no explicit mention of a "training set" for an AI/ML model, this question is not directly applicable. For the development of the immunoassay itself, the "ground truth" during development would be established through a rigorous process of biochemical characterization, optimization using known concentrations of analytes, and comparisons to established reference methods or highly purified standards to ensure accurate and specific binding. This is an engineering/biochemical development process rather than establishing a "ground truth" for a dataset in the AI/ML sense.

Summary

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K030655

510(K) Summary Axis-Shield HoloTC RIA

Summary of Safety and Effectiveness Information Supporting a Substantially Equivalent Determination

Statement of Intended Use

The Axis-Shield HoloTC RIA is an in-vitro diagnostic assay for quantitative mcasurement of holotranscobalamin (vitamin B12 bound to transcobalamin) in human serum or plasma. Measurements obtained by this device are used in the diagnosis and treatment of vitamin B12 deficiency. HoloTC RIA is calibrated with HoloTC Calibrators. HoloTC controls are assayed for the verification of the accuracy and precision of the HoloTC RIA.

Summary of Technological Characteristics

1. The use of a transcobalamin specific antibody, this allows the quantitation of only the cobalamin bound to the protein transcobalamin as opposed to measurement of cobalamin bound to all proteins in the predicate device and
1. The detection signal is radioactivity (37Co) as opposed to chemiluminescence in the predicate device.

Method Comparison

The Axis-Shield HoloTC RIA was compared to the Bayer Advia Centaur VB12 assay (K993571) using 392 patient samples with vitamin B12 concentrations ranging from 114-821 pmol/L.

Linear regression (least squares) vielded the following statistics:

\HoloTC pmol/L = 0.55\VB12-2 pmol/L r2 = 0.52

Reference Interval

Based on a Finnish population of normal individuals (n=303, age 22-88 years) the 95% central reference interval was found to be 37-171 pmol/L.

Analysis of covariance demonstrated that HoloTC levels depended on gender (but not age) in this reference population. The 90 % confidence intervals for the lower limit of the reference range are for the whole population, males and females 36-37 pmol/L, 37-39 pmol/L and 35-36 pmol/L, respectively.

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External Evaluation

The Axis-Shield HoloTC RIA external laboratory performance was evaluated at two external study sites, UK and Denmark. Performance of the system was demonstrated by using three different levels of native and pooled serum samples in addition to kit controls Low and High. CV for the three serum samples were within the given acceptance criteria, <15 %.

Precision

Precision studies were done according to NCCLS standard. Precision of the system was demonstrated by using three levels of serum samples, low, medium and high (14, 67 and 139 pmol/L HoloTC, respectively). Within-run CV for duplicate measurements of serum low, medium and high were 11%, 5% and 8%, respectively and total precision was 11%, 6% and 8%, respectively.

Linearity

Dilution Linearity within the working range of 10-160 pmol/L was demonstrated to meet all criteria for linearity. Recovery test of HoloTC spiked serum samples was demonstrated to be within acceptance criteria. Dilution of high HoloTC sample into six dilutions showed a correlation coefficient r2 = 0.998, slope = 1.01 ± 0.02 and y-intercept = - 5 ± 2 pmol/L for dilutions ranging between 13 - 182 pmol/L HoloTC. Limit of Quantification (LOQ) and Limit of Detection (LOD) were demonstrated to be 8-176 pmol/L and 6 pmol/L, respectively. Cross-reactivity was tested against haptocorrin (HC) and cross-reactivity was well within the acceptance criteria of ±10% for up to 70,000pg/mL HC spiked serum samples.

Interference

Effect on quantification by endogenous interfering substances was tested. billirubin, haemoglobin, total protein and lipids showed less that 10% interference.

Serum/EDTA Plasma comparison

50 paired serum and EDTA plasma samples collected from the same donors were analysed and compared. No significant difference in measured HoloTC level was demonstrated between serum sample and EDTA sample. In-use stability of the reagent and calibrator kit showed stability up to three months after opening.

Sensitivity and Specificity

The estimated concordance and relative sensitivity and specificity of HoloTC RIA was calculated using samples from 3 study sites that were classified as being likely (n=112) and not likely (n=313) vitamin B12 deficient based on cobalamin and MMA levels. A general parametric approach was used. The estimated concordance, sensitivity, and specificity for each study was weighted and then all were combined to give the following estimates; concordance 80%, sensitivity 99.5%, specificity 76.3%.

Conclusion

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In conclusion these data demonstrate that the Axis-Shield HoloTC RIA is as safe and effective as, and is substantially equivalent to, the Bayer Advia Centaur VB12 assay.

Contact information:

Axis-Shield C/O - R & R REGISTRATIONS Ronald G. Leonardi President San Diego, CA 92131 858-586-0751 858-586-1108 (fax)

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DEPARTMENT OF HEALTH & HUMAN SERVICES

Image /page/3/Picture/1 description: The image shows the logo for the Department of Health & Human Services (USA). The logo features a stylized abstract design with three wave-like shapes, possibly representing people or services. The text "DEPARTMENT OF HEALTH & HUMAN SERVICES USA" is arranged in a circular pattern around the abstract design.

Public Health Service

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

JAN 2 9 2004

Axis-Shield Biochemicals, ASA c/o Ronald G. Leonardi, Ph. D. President R & R Registrations P.O. Box 262069 San Diego, CA 92196-2069

K030655 Re:

Trade/Device Name: Axis-Shield Holo TC RIA Regulation Number: 21 CFR 862.1810 Regulation Name: Vitamin B12 test system Regulatory Class: Class II Product Code: CDD; JIS; JJX Dated: November 4, 2003 Received: November 5, 2003

Dear Dr. Leonardi:

We have reviewed your Section 510(k) premarket notification of intent to market the device we nave reviewed your betermined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate rol use stated in the encreat date of the Medical Device Amendments, or to commerce proc to may 20, 2011 - 12:00 accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). and Cosmetic For ( rol) lies the device, subject to the general controls provisions of the Act. The I ou may, meroloro, manes of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. 1

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it If your device is elabilition (toontrols. Existing major regulations affecting your device can may be subject to sail adaxt Federal Regulations (CFR), Parts 800 to 895. In addition, FDA be found in Ther 21, 0 wouncements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean I loast be advised that I Drivian that your device complies with other requirements of the Act that I Dr has Intact and regulations administered by other Federal agencies. You must of any I catal statutes and states and including, but not limited to: registration and listing (21 comply with an and and the Parts 801 and 809); and good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820).

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Page 2

This letter will allow you to begin marketing your device as described in your Section 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 information about the application of labeling requirements to your device, or questions on the promotion and advertising of your device, please contact the Office of In Vitro Diagnostic Device Evaluation and Safety at (301) 594-3084. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21CFR Part 807.97). 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) 443-6597 or at its Internet address http://www.fda.gov/cdrh/dsma/dsmamain.html.

Sincerely yours,

Jean M. Cooper, US, DVM.

Yean M. Cooper, MS, D.V.M. Director Division of Chemistry and Toxicology Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health

Enclosure

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X030655

510(k) Number (if known)

Device Name:Axis-Shield HoloTC RIA

INDICATIONS FOR USE:

The Axis-Shield HoloTC RIA is an in-vitro diagnostic assay for quantitative measurement of the fraction of cobalamin (vitamin B12) bound to the carrier protein transcobalamin in human serum or plasma. Measurements obtained by this device are used in the diagnosis and treatment of vitamin B12 deficiency.

(PLEASE DO NO 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)

OR Over-The-Counter Use

(Optional Format 1-2-96)

Carol C. Benson for Jean Cooper, OhiM

(Division Sign-Off) Division of Clinical Laboratory De 510(k) Number -

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.