For the quantitative determination of folate in serum or EDTA plasma and red blood cells using the Folate BA (Biotin Avidin) assay on the Chiron Diagnostics ACS: 180® Automated Chemiluminescence Systems.

Device Description

The Chiron Diagnostics ACS:180 Folate assay is a competitive immunoassay using direct chemiluminescent technology. Folate in the patient sample competes with acridinium esterlabeled folate in the Lite Reagent for a limited amount of biotin-labeled folate binding protein. Biotin-labeled folate binding protein binds to avidin which is covalently coupled to paramagnetic particles in the Solid Phase. In the ACS:180 Folate assay the sample is pretreated to release the folate from endogenous binding proteins in the sample. The system performs the following steps for calibrators, quality control samples, and patient samples: dispenses 150 uL of sample into a cuvette, dispenses 50 µL of DTT, dispenses 100 µL of folate binding protein and 200 µL of Solid Phase and incubates for 5.0 minutes at 37°C, dispenses 100 µL of Lite Reagent and incubates for 2.5 minutes at 37°C, separates, aspirates, and washes the cuvettes with reagent water, dispenses 300 uL each of Reagent 1 and Reagent 2 to initiate the chemiluminescent reaction, reports results according to the selected option, as described in the system operating instructions or in the online help system. An inverse relationship exists between the amount of folate present in the patient sample and the amount of relative light units (RLUs) detected by the system.

AI/ML Overview

Here's an analysis of the provided text, focusing on the acceptance criteria and the study details:

Device Name: Chiron Diagnostics ACS: 180 Folate Assay
Intended Use: For the quantitative determination of folate in serum or EDTA plasma and red blood cells using the Folate BA (Biotin Avidin) assay on the Chiron Diagnostics ACS:180® Automated Chemiluminescence Systems.

1. Table of Acceptance Criteria and Reported Device Performance

The document does not explicitly state "acceptance criteria" with pass/fail thresholds. Instead, it presents "Performance Characteristics" which indicate the expected performance of a well-functioning assay. I will interpret these performance characteristics as the de-facto acceptance criteria for the device, given they are presented as a summary of safety and effectiveness.

Performance Characteristic	Acceptance Criteria (Implied)	Reported Device Performance
Sensitivity / Minimum Detectable Concentration	A minimum detectable concentration (lower limit of quantitation) should be established.	0.25 ng/mL (0.6 nmol/L)
Assay Reportable Range	The assay should accurately measure folate concentrations up to a defined upper limit.	Up to 20 ng/mL (45.4 nmol/L)
Method Comparison (Serum)	A strong correlation (r) should exist with an alternate folate assay, with a slope close to 1 and intercept close to 0.	r = 0.95ACS:180 Folate = 0.92 (alternate chemiluminescent method) + 0.21 ng/mL
Method Comparison (RBC)	A strong correlation (r) should exist with an alternate folate assay, with a slope close to 1 and intercept close to 0.	r = 0.96ACS:180 RBC Folate = 0.93 (alternate chemiluminescent method) + 52.8 ng/mL
Precision (Within-run %CV)	Acceptable variability within a single run should be demonstrated across different folate concentrations.	4.88% - 7.95%
Precision (Total %CV)	Acceptable total variability should be demonstrated across different folate concentrations, lots, runs, and systems.	5.36% - 9.24%
Expected/Normal Range (Serum)	Reference ranges for normal and deficient populations should be established.	Normal: 4.25-23.8 ng/mL (9.65-54.0 nmol/L)Deficient: 0.0-2.31 ng/mL (0.0-5.24 nmol/L)
Expected/Normal Range (RBC)	Reference ranges for normal and deficient populations should be established.	Normal: 322-886 ng/mL (731-2011 nmol/L)Deficient: 9-157 ng/mL (20.4-356 nmol/L)

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

Sensitivity/Analytical Sensitivity: 20 replicate determinations of the folate zero standard in 7 assays with 3 lots of reagents. (Total of 20 x 7 x 3 = 420 measurements, but focused on the zero standard.) The country of origin for this data is not specified, nor is whether it was retrospective or prospective, but it implies a controlled laboratory study.
Method Comparison (Serum): 258 serum samples.
Method Comparison (RBC): 189 red blood cell samples.
Expected Results/Reference Range (Serum): 263 serum samples, described as "apparently healthy males and females".
Expected Results/Reference Range (RBC): 109 red blood cell samples, described as "apparently healthy males and females".
Precision: Four samples were assayed six times with three lots of reagents in 23 runs on four systems (n = 138 for each sample). (Total of 4 samples * 6 replicates * 23 runs = 552 sample measurements; or, 4 samples * 138 measurements per sample = 552 total measurements).

Data Provenance: The document states that "The data was obtained on apparently healthy males and females from the United States" for the Expected Results (reference ranges). For other sections (e.g., method comparison, precision, sensitivity), the country of origin and design (retrospective/prospective) are not explicitly stated but are implied to be from a controlled laboratory setting (likely prospective) given the nature of the tests.

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

The document does not mention the use of experts to establish ground truth for the test set in the traditional sense of consensus or adjudication for diagnostic imaging studies.

However, for the deficient categories in the "Expected Results" table, the diagnosis was established based on "bone and/or peripheral blood smear pathology and other criteria including: megaloblastic anemia, folate deficient diet, malabsorption, alcoholism, Tropical Sprue, abnormal blood parameters including mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), and hematocrit (HCT)." While this indicates a clinical diagnosis process, it doesn't specify if a panel of experts was used or their qualifications for this specific study. It implies standard diagnostic practices were followed.

4. Adjudication Method for the Test Set

No explicit adjudication method (e.g., 2+1, 3+1) is mentioned for any of the studies described. The ground truth for deficient samples appears to rely on a set of clinical and pathological criteria rather than expert consensus on individual cases specifically for the study.

5. If a Multi Reader Multi Case (MRMC) Comparative Effectiveness Study Was Done

No. This document describes the performance characteristics of an in vitro diagnostic (IVD) assay (a lab test), not an AI-assisted diagnostic imaging device. Therefore, a multi-reader, multi-case (MRMC) comparative effectiveness study with human readers improving with AI vs without AI assistance is not applicable and was not performed.

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

Yes, this is a standalone performance study. The document details the analytical performance of the ACS:180 Folate assay system (an automated chemiluminescence system) itself, independent of human interpretation of its results. The reported metrics (sensitivity, reportable range, method comparison, precision, expected ranges) are all inherent to the device's function.

7. The Type of Ground Truth Used

Analytical Sensitivity, Reportable Range, Precision: Ground truth is established by the known concentrations of calibrated standards and controls used in the assay.
Method Comparison: Ground truth is functionally established by an "alternate chemiluminescent method" (the predicate or another established assay), rather than an absolute gold standard. The aim is to show agreement with an existing method.
Expected Results (Normal/Deficient Ranges):
- Normal: Derived from samples obtained from "apparently healthy males and females". The assumption is that these individuals represent a "normal" folate status.
- Deficient: Diagnosed by "bone and/or peripheral blood smear pathology and other criteria including: megaloblastic anemia, folate deficient diet, malabsorption, alcoholism, Tropical Sprue, abnormal blood parameters including mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), and hematocrit (HCT)." This is clinical diagnostic ground truth.

8. The Sample Size for the Training Set

The document does not describe a "training set" as it would for a machine learning algorithm. This is a conventional IVD assay, not an AI/ML-based device in the modern sense. The "training" in this context would refer to internal development and optimization by the manufacturer, for which sample sizes are not typically reported in such summaries.

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

As stated above, the concept of a "training set" and its associated ground truth establishment is not relevant in the context of this traditional IVD assay and its regulatory submission. The device's performance is characterized against known analytical standards and clinical classifications, not by 'learning' from a labeled dataset.

Summary

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JUN 30 1999

Company Confidential

K991582

Summary of Safety and Effectiveness

As required by 21 CFR 807.92, the following 510(k) Summary is provided:

1. Submitters Information

Contact person:	William J. PignatoDirector of Regulatory Affairs
Address:	Chiron Diagnostics Corporation63 North StreetMedfield, MA 02052
Phone:Faxe-mail	(508) 359-3825(508) 359-3356william.pignato.b@bayer.com

April 30, 1999 Date Summary Prepared:

2. Device Information

Proprietary Name:	Chiron Diagnostics ACS: 180 Folate
Common Name:	Folate Immunological test system
Device Classification:	Class II

3. Predicate Device Information

Name:	Chiron Diagnostics ACS: 180 Folate Immunoassay
Manufacturer:	Chiron Diagnostics Corporation

4. Device Description

Folate, with vitamin B12, is essential for DNA synthesis, which is required for normal red blood cell maturation. Humans obtain folate from dietary sources including fruits, green and leafy vegetables, yeast, and organ meats. Folate is absorbed through the small intestine and stored in the liver.

Low folate intake, malabsorption as a result of qastrointestinal diseases, pregnancy, and drugs such as phenytoin are causes of folate deficiency. Folate deficiency is also associated with chronic alcoholism. Folate and vitamin B12 deficiency impair DNA synthesis, causing macrocytic anemias. These anemias are characterized by abnormal maturation of red blood cell precursors in the bone marrow, the presence of megaloblasts, and decreased red blood cell survival.

Since both folate and vitamin B12 deficiency can cause macrocytic anemia, appropriate treatment depends on the differential diagnosis of the deficiency. Serum folate measurement provides an early index of folate status. However, folate is much more concentrated in red blood cells than in serum so the red blood cell folate measurement

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more closely reflects tissue stores. Red blood cell folate concentration is considered the most reliable indicator of folate status.

5. Statement of Intended Use

6. Summary of Technological Characteristics

The system performs the following steps for calibrators, quality control samples, and patient samples:

dispenses 150 uL of sample into a cuvette

dispenses 50 µL of DTT

dispenses 100 µL of folate binding protein and 200 µL of Solid Phase and incubates for 5.0 minutes at 37°C

dispenses 100 µL of Lite Reagent and incubates for 2.5 minutes at 37°C

separates, aspirates, and washes the cuvettes with reagent water6

dispenses 300 uL each of Reagent 1 and Reagent 2 to initiate the chemiluminescent reaction

reports results according to the selected option, as described in the system operating instructions or in the online help system

An inverse relationship exists between the amount of folate present in the patient sample and the amount of relative light units (RLUs) detected by the system.

6. Performance Characteristics

Expected Results

To determine the reference range for the ACS:180 Folate assay for serum and RBC folate, data was obtained on 263 serum and 109 red blood cell folate samples, respectively. The normal ranges are based on 95% confidence intervals and the deficient ranges represent the observed ranges.

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Company Confidential

Category	N	Mean(ng/mL)	Range(ng/mL)	Mean(nmol/L)	Range(nmol/L)
Serum
folate
deficient*	32	1.08	0.0-2.31	2.45	0.0-5.24
normal	231	9.89	4.25-23.8	22.4	9.65-54.0
RBC
folate
deficient*	10	64.4	9-157	146	20.4-356
normal	99	545	322-886	1237	731-2011

Diagnosed by bone and/or peripheral blood smear pathology and other criteria including:

megaloblastic anemia
. folate deficient diet
malabsorption
alcoholism ●
. Tropical Sprue
. abnormal blood parameters including mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), and hematocrit (HCT).

Laboratories should consider these reference ranges as guidelines only. The data was obtained on apparently healthy males and females from the United States. Due to population demographic factors, assay methods, calibration, and reagent specificity, each laboratory should establish its own reference ranges for the diagnostic evaluation of patient results.

Sensitivity and Assay Reportable Range

The ACS:180 Folate assay measures folate concentrations up to 20 ng/mL (45.4 nmol/L) with a minimum detectable concentration of 0.25 ng/mL (0.6 nmol/L). Analytical sensitivity is defined as the concentration of folate that corresponds to the RLUs that are two standard deviations less than the mean RLUs of 20 replicate determinations of the folate zero standard in 7 assays with 3 lots of reagents.

Method Comparison

For 258 serum samples in the range of 0 to 20 ng/mL (0 to 45.4 nmol/L), the relationship between the ACS:180 Folate assay and an alternate folate assay is described by the equation:

ACS:180 Folate = 0.92 (alternate chemiluminescent method) + 0.21 ng/mL Correlation coefficient (r) = 0.95

For 189 red blood cell samples in the range of 9.0 to 882 ng/mL (20.4 to 2002 nmoV/L), the relationship between the ACS:180 Folate assay and an alternate folate assay is described by the equation:

ACS:180 RBC Folate = 0.93 (alternate chemiluminescent method) + 52.8 ng/mL Correlation coefficient (r) = 0.96

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Precision

Four samples were assayed six times with three lots of reagents in 23 runs on four systems (n = 138 for each sample), over a period of three days. The following results were obtained:

Mean Folate (ng/mL)	Mean Folate (nmol/L)	Within-run % CV	Total % CV
1.91	4.34	7.95	9.24
5.94	13.5	5.36	8.79
10.6	24.1	5.61	6.60
15.4	35.0	4.88	5.36

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Image /page/4/Picture/1 description: The image shows the seal of the Department of Health & Human Services (HHS). The seal features a stylized eagle with three lines forming its body and wings. The words "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" are arranged in a circular pattern around the eagle.

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

JUN 30 1999

Mr. William J. Pignato Director of Regulatory Affairs Chiron Diagnostics Corporation 63 North Street Medfield, Massachusettes 02052-1688

K991582 Re:

Trade Name: Chiron Diagnostics ACS: 180® Folate Assay Regulatory Class: II Product Code: CGN, JIS Dated: April 30, 1999 Received: May 7, 1999

Dear Mr. Pignato:

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 QS 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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Page 2

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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Company Confidential

Page | of |

510(k) Number (if known): K991682

Device Name: Chiron Diagnostics ACS:180 Folate Assay

Indications for Use:

Dean Cooper
(Division Sign-Off)

vision of Clinical Lab ratory Devices 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

§ 862.1295 Folic acid test system.

(a)
Identification. A folic acid test system is a device intended to measure the vitamin folic acid in plasma and serum. Folic acid measurements are used in the diagnosis and treatment of megaloblastic anemia, which is characterized by the presence of megaloblasts (an abnormal red blood cell series) in the bone marrow.(b)
Classification. Class II.