This microwell enzyme linked immunoabsorbant assay (ELISA) detection kit is an in vitro diagnostic (IVD) immunoassay for the detection of Cryptosporidium antigen in human feces using peroxidase as the enzyme. The assay may be read visually or with an ELISA reader. This IVD assay is intended to be used with stools that are fresh, frozen or preserved in 10% formalin, SAF or Medical Chemical Corporation's (MCC) Universal fixative. Concentrated samples cannot be used with this IVD.

Device Description

This microwell enzyme-linked immunoabsorbant assay (ELISA) detection kit (Cryptosporidium ELISA Kit) is an in vitro diagnostic (IVD) immunoassay for the detection of antigen to Cryptosporidium species in human feces using peroxidase as the indicator enzyme. The assay may be read visually or with an ELISA reader. Concentrated fecal samples cannot be used with this immunoassay. Rather, this IVD Cryptosporidium ELISA Kit is intended to be used with human stools that are fresh, frozen or preserved in 10% formalin, SAF or Medical Chemical Corporation's (MCC's) Universal fixative in a clinical laboratory use setting.

This ELISA is an in vitro immunoassay for the qualitative determination of Cryptosporidium species antigen in feces. The ELISA uses a rabbit anti-Cryptosporidium antibody to capture the antigen from the stool supernatant. A second goat anti-Cryptosporidium antibody is then added which sandwiches the captured antigen. This reaction is visualized by the addition of an anti-second antibody conjugated to peroxidase and the chromogen tetramethylbenzidine (TMB). The resulting blue color development indicates the presence of Cryptosporidium species antigens being bound by the anti-Cryptosporidium antibodies.

AI/ML Overview

Here's an analysis of the acceptance criteria and study details for the IVD Research Inc.'s Cryptosporidium Human Fecal Antigen Detection Microwell ELISA Kit, based on the provided text:

Acceptance Criteria and Device Performance

The acceptance criteria are implied by the performance metrics reported for comparison to the gold standard and the predicate device. For diagnostic tests, this generally includes high sensitivity, specificity, and agreement rates.

Performance Metric	Acceptance Criteria (Implied)	Reported Device Performance
Study #1 (Outside Lab) vs. O&P Microscopy	Sensitivity ≥ 90% (e.g., comparable to predicate and gold standard)	Sensitivity: 100% (24/24)
	Specificity ≥ 90% (e.g., comparable to predicate and gold standard)	Specificity: 97% (146/150) (95% CI: 93% to 99%)
Study #2 (Manufacturer Performed) vs. Commercial ELISA	Positive Agreement ≥ 90% (e.g., similar to predicate)	Positive Agreement: 100% (14/14)
	Negative Agreement ≥ 90% (e.g., similar to predicate)	Negative Agreement: 100% (30/30)
Analytical Sensitivity	Detection of Cryptosporidium antigen at relevant clinical levels	Approximately 50 nanograms of soluble protein per ml of Cryptosporidium species antigen or 10 oocysts per well (OD range of 0.15 to 0.30)
Cross-Reactivity	No significant cross-reactivity with common fecal pathogens	No cross-reactions seen with a detailed list of 24 other organisms (parasites, bacteria) and 29 additional bacterial species.

Study Details

Sample Size used for the test set and the data provenance:
- Study #1 (Outside Lab): 174 human fecal specimens (formalin, SAF, fresh/frozen and Universal Fixative preserved stools). Data provenance is from "clinical laboratories or in-house collections," making it retrospective. The country of origin is not explicitly stated but is implied to be the US, given the FDA submission.
- Study #2 (Manufacturer Performed): 44 fresh or fresh/frozen human fecal specimens. Data provenance is from "clinical laboratories or in-house collections," making it retrospective. The country of origin is implied to be the US.
Number of experts used to establish the ground truth for the test set and the qualifications of those experts:
- The document does not specify the number of experts or their qualifications for establishing the ground truth (O&P microscopy results or other commercial ELISA results). It only refers to "O&P microscopy" as the "gold standard for parasitology" and "another commercial ELISA."
Adjudication method (e.g., 2+1, 3+1, none) for the test set:
- The document does not describe any specific adjudication method for the ground truth. It simply states that the results were interpreted against O&P microscopy or another commercial ELISA.
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:
- No, a multi-reader multi-case (MRMC) comparative effectiveness study was not performed. This is an ELISA kit, not an AI-assisted diagnostic tool for human readers.
If a standalone (i.e., algorithm only without human-in-the-loop performance) was done:
- Yes, this is effectively a standalone diagnostic device. The ELISA kit itself provides the qualitative determination. While a human reads the results (visually or with an ELISA reader), the performance metrics reported are for the kit's ability to detect the antigen, independent of human interpretive variability beyond simply reading a color change or optical density.
The type of ground truth used (expert consensus, pathology, outcomes data, etc.):
- For Study #1, the ground truth was O&P (Ova and Parasite) microscopy, considered the "gold standard for parasitology."
- For Study #2, the ground truth was O&P microscopy and/or another commercial ELISA.
The sample size for the training set:
- The document does not explicitly mention a "training set" in the context of machine learning or algorithm development. This refers to an ELISA kit, not an AI model. The studies described are validation (test) sets.
How the ground truth for the training set was established:
- Given that this is an immunoassay kit and not an AI/machine learning device, there isn't a "training set" in the conventional sense. The "ground truth" for the test sets was established by O&P microscopy or a commercial ELISA, as described above.

Summary

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JUL 1 0 2003

510(k) Summary of Safety and Effectiveness for IVD Research Inc.'s Cryptosporidium Human Fecal Antigen Detection Microwell ELISA Kit

Submitter:	IVD Research Inc.5909 Sea Lion Place, Suite DCarlsbad, CA 92008 USATel: 760-929-7744 Contact: Dave Lambillotte, President
Prepared:	12 May 2003

Common name: Cryptosporidium Human Fecal Antigen Detection Microwell ELISA Kit

Proprietary name: IVD Research's Cryptosporidium Human Fecal Antigen Detection Microwell ELISA Kit

Classification name: Entamoeba histolytica serological reagents

Predicate Device: LMD Laboratories' Cryptosporidium Antigen Detection ELISA, 510(k) K955345

DESCRIPTION AND PRINCIPLE:

This ELISA corresponds to FDA Classification Name: Entamoeba Histolytica Serological Reagent, a class II (non-exempt) Device, within the Microbiology Classification Panel, having FDA Reg. Citation Number: 21 CFR 866.3220, and FDA Product Codes: MHJ.

SUBSTANTIAL EQUIVALENCE COMPARISON

A similar ELISA kit was developed and manufactured by LMD Laboratories (San Diego, CA). The FDA cleared this assay under 510(k) K955345 in 7 June 1996. This current kit uses the same methodology (ELISA) and similar reagents as that cleared predicate assay.

Following is the data that shows this firm's ELISA provides substantial equivalence to the gold standard for parasitology (O&P microscopy) as well as the predicate device. The 95% Confidence Interval (CI) was determined using a Confidence Interval Analysis Microcomputer Program published by the British Medical Journal.

All specimens are human samples supplied by clinical laboratories or in-house collections.

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Study #1 (Outside Lab)

A total of 174 formalin, SAF, fresh/frozen and Universal Fixative preserved stools were tested against the ELISA test The results were interpreted visually.

The following results were obtained.

	Micro + (O&P)	Micro - (O&P)
IVDELISA +	24	4
IVDELISA -	0	146
95% CI = 86% to 100%

Sensitivity: 100% (24/24) Specificity: 97% (146/150) 95% CI = 93% to 99%

No cross-reactions were seen with the following organisms:

Entamoeba hartmanni, Endolimax nana, Entamoeba histolytica/dispar, Entamoeba coli, Blastocystis hominis, Dientamoeba fragilis, Chilomastix mesnili, Giardia lamblia, Cyclospora cayetanensis oocysts, Strongyloides stercoralis, Ascaris lumbricoides, Enterobius vermicularis (pinworm), Diphyllobothrium species, Hymenolepis nana, Clonorchis sinensis, Enteromonas hominis, Trichuris trichiura, Iodamoeba buetschlii, Hookworm (Necator americanus), Schistosoma mansoni, Taenia eggs, Isospora belli, Entamoeba polecki, adenovirus, E. coli, Campylobacter, Salmonella, Shigella, Yersinia and 29 other bacterial species (list available on request).

Study #2 (Manufacturer Performed)

A total of 44 fresh or fresh/frozen stools examined by O&P microscopy and/or another commercial ELISA were tested against this IVD ELISA. The following results were obtained.

	ELISA	ELISA
IVDELISA +	14	0
IVDELISA -	0	30

Positive Agreement = 100% (14/14) Negative Agreement = 100% (30/30)

The breakdown for the samples in the studies is as follows:

Media	Cryptosporidium +	Other Parasites
Fresh/Frozen	14	0
10% Formalin	24	25
SAF	1	83
MCC	2*	31

*These two samples were seeded samples.

Analytical Sensitivity

This assay can detect approximately 50 nanograms of soluble protein per ml of Cryptosporidium species antigen or 10 oocysts per well. At these detection levels, an OD range of 0.15 to 0.30 can be expected.

CONCLUSION

IVD Research's Cryptosporidium Human Fecal Antigen Detection Microwell ELISA Kit uses similar methodology to the predicate device. In testing of various fecal specimens, the assay also showed equivalent sensitivity and specificity to the predicate device and O&P microscopy.

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DEPARTMENT OF HEALTH & HUMAN SERVICES

Image /page/2/Picture/1 description: The image shows the logo for the U.S. Department of Health & Human Services. The logo is circular and contains the words "DEPARTMENT OF HEALTH & HUMAN SERVICES · USA" around the perimeter. Inside the circle is an abstract symbol that resembles an eagle or a bird in flight, composed of three curved lines.

JUL 1 0 2003

Food and Drug Administration 2098 Gaither Road Rockville MD 20850

IVD Research, Inc. c/o Alfredo J. Quattrone, Ph.D., D.A.B.T. California Department of Health Food & Drug Branch P.O. Box 942732 (MS-357) Sacramento, CA 94234

Re: K031059

Trade/Device Name: Cryptosporidium Human Fecal Antigen Detection Microwell ELISA Kit

Regulation Number: 21 CFR 866.3220 Regulation Name: Entamoeba histolytica serological reagents Regulatory Class: Class II Product Code: MHJ Dated: March 27, 2003 Received: March 28, 2003

Dear Dr. Quattrone:

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. Iisting 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 Title 21, Code of Federal 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 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 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,

Steven Sutman

Steven I. Gutman, M.D., M.B.A. Director Office of In Vitro Diagnostic Device Evaluation and Safety Center for Devices and Radiological Health

Enclosure

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INDICATIONS FOR USE STATEMENT

510(k) Number:	Pending this Submission K031059
Device Name:	Cryptosporidium Human Fecal Antigen DetectionMicrowell ELISA Kit
Indications for Use:	This microwell enzyme linked immunoabsorbant assay (ELISA)detection kit is an in vitro diagnostic (IVD) immunoassay for thedetection of Cryptosporidium antigen in human feces usingperoxidase as the enzyme. The assay may be read visually or withan ELISA reader. This IVD assay is intended to be used withstools that are fresh, frozen or preserved in 10% formalin, SAF orMedical Chemical Corporation's (MCC) Universal fixative.Concentrated samples cannot be used with this IVD.

510(k) Number:

Pending this Submission K031059

Device Name:

Cryptosporidium Human Fecal Antigen DetectionMicrowell ELISA Kit

Indications for Use:

This microwell enzyme linked immunoabsorbant assay (ELISA)detection kit is an in vitro diagnostic (IVD) immunoassay for thedetection of Cryptosporidium antigen in human feces usingperoxidase as the enzyme. The assay may be read visually or withan ELISA reader. This IVD assay is intended to be used withstools that are fresh, frozen or preserved in 10% formalin, SAF orMedical Chemical Corporation's (MCC) Universal fixative.Concentrated samples cannot be used with this IVD.

(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) OR

Over-The-Counter Use

(Optional Format 1-2-96)

Freddie Hi. Poole

Division Sign-Off

Office of In Vitro Diagnostic Device Evaluation and Safety

510(k) K031059

Regulation Number and Section

§ 866.3220

Entamoeba histolytica serological reagents.(a)
Identification. Entamoeba histolytica serological reagents are devices that consist of antigens and antisera used in serological tests to identify antibodies toEntamoeba histolytica in serum. Additionally, some of these reagents consist of antisera conjugated with a fluorescent dye (immunofluorescent reagents) used to identifyEntamoeba histolytica directly from clinical specimens. The identification aids in the diagnosis of amebiasis caused by the microscopic protozoan parasiteEntamoeba histolytica and provides epidemiological information on diseases caused by this parasite. The parasite may invade the skin, liver, intestines, lungs, and diaphragm, causing disease conditions such as indolent ulcers, an amebic hepatitis, amebic dysentery, and pulmonary lesions.(b)
Classification. Class II (special controls). The device is exempt from the premarket notification procedures in subpart E of part 807 of this chapter subject to § 866.9.