TREND CRYPTOSPORIDIUM DETECTION TEST SYSTEMS

K955755 · Trend Scientific, Inc. · MHJ · Jul 11, 1996 · Microbiology

Device Facts

• Record ID: K955755
• Device Name: TREND CRYPTOSPORIDIUM DETECTION TEST SYSTEMS
• Applicant: Trend Scientific, Inc.
• Product Code: MHJ · Microbiology
• Decision Date: Jul 11, 1996
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 866.3220
• Device Class: Class 2

Indications for Use

The intended use of the device is for the qualitative determination of Cryptosporidium antigen in feces. The ELISA test kit is indicated for use with fecal specimens from patient’s with diarrhea to aid in the detection of Cryptosporidium gastrointestinal infection.

Device Story

The TREND Cryptosporidium Direct Detection Test System is an antigen capture ELISA for fecal specimens. It utilizes microplate wells coated with antibodies to capture Cryptosporidium antigens from stool supernatant. A second anti-Cryptosporidium antibody creates a sandwich complex, followed by an HRP-conjugated anti-immunoglobulin antibody to amplify the signal. A substrate is added to produce a blue color, which turns yellow upon addition of a stop solution. Results are interpreted spectrophotometrically via a microplate reader or visually. The device is intended for use in clinical laboratories to assist in diagnosing Cryptosporidium infection, which causes symptoms like diarrhea, nausea, and abdominal cramps, particularly in immunocompromised patients. The test provides qualitative detection of the parasite, aiding clinical decision-making regarding patient management.

Clinical Evidence

Bench testing performed at three locations (in-house, parasitology reference lab, university research center). Performance validated against conventional microscopy with modified acid fast (MAF) staining. Study A (n=96) showed 96.2% sensitivity and 97.1% specificity; Study B (n=97) showed 97% sensitivity and 100% specificity. Analytical sensitivity determined at 4.6 ng/ml. Cross-reactivity testing performed against 26 parasitic organisms and various enteric bacteria, fungi, and viruses with no cross-reactivity observed. Precision and reproducibility testing conducted at three sites.

Technological Characteristics

Antigen capture ELISA; microplate-based. Detection via HRP-conjugated antibody and colorimetric substrate (blue/yellow). Analytical sensitivity: 4.6 ng/ml. Compatible with spectrophotometric microplate readers. In vitro diagnostic test kit.

Indications for Use

Indicated for patients with diarrhea to aid in the detection of Cryptosporidium gastrointestinal infection.

Regulatory Classification

Identification

Entamoeba histolytica serological reagents are devices that consist of antigens and antisera used in serological tests to identify antibodies to Entamoeba histolytica in serum. Additionally, some of these reagents consist of antisera conjugated with a fluorescent dye (immunofluorescent reagents) used to identify Entamoeba histolytica directly from clinical specimens. The identification aids in the diagnosis of amebiasis caused by the microscopic protozoan parasite Entamoeba 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.

Special Controls

*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.

Submission Summary (Full Text)

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JUL 11 1996
K955755

Page 2 of 4

Summary of 510(k) Safety and Effectiveness information.
Trade / Proprietary Name: TREND Cryptosporidium Direct Detection
Test System, Cat.# KA49-

## DESCRIPTION and INTENDED USE of the DEVICE:

The intended use of the device is for the qualitative determination of *Cryptosporidium* antigen in feces. The ELISA test kit is indicated for use with fecal specimens from patient’s with diarrhea to aid in the detection of *Cryptosporidium* gastrointestinal infection.

Traditional detection of parasitic organisms has been conventional microscopic examination of fecal material. During the last few years, ELISA technology for the identification of specific parasites has become more common and cost effective for many laboratories.

*Cryptosporidium parvum* is a coccidian parasite recognized as a human and animal pathogen. Symptoms of a parasitemia caused by this organism include diarrhea, nausea, low grade fever, abdominal cramps, and anorexia. The organism is a common cause of self-limiting diarrhea in immunocompetent persons and may cause a chronic, severe, and life-threatening diarrhea in immunocompromised patients, particularly those with HIV. Transmission modes include person-to-person or animal-to-person contact with fecal material and contact with contaminated food or water supplies.

## SCIENTIFIC PRINCIPLES:

The device is an antigen capture enzyme linked immunosorbent assay (ELISA) for use with stools/ fecal material. The antigen capture takes place in microplate wells. During the first incubation, *Cryptosporidium* antigens present in the stool supernatant are captured by antibodies attached to the test wells. The second incubation adds an additional anti-*Cryptosporidium* antibody that "sandwiches" the antigen. The next incubation identifies the antibody/ antigen complex and amplifies the signal by the addition of an anti-immunoglobulin antibody conjugated to horse radish peroxidase (HRP). After washings that remove unbound enzyme, a substrate is added which develops a blue color in the presence of the enzyme complex. The stop solution ends the reaction and turns the blue color to yellow. The results may be read spectrophotometrically with a microplate reader or visually.

## TECHNOLOGY:

The ELISA technology of the device, as defined in the Scientific Principles section above, is utilized in a number of in vitro diagnostic test kits currently used in diagnostic laboratories. ELISA kits specific for the detection of other parasitic organisms (for example, *Giardia lamblia*) are also being used.

0496

{1}

Summary of 510(k) Safety and Effectiveness information.
Trade / Proprietary Name: TREND Cryptosporidium Direct Detection
Test System, Cat. # KA49-

## PERFORMANCE TESTING:

Clinical Laboratory Bench Studies were performed in-house and at two off-site locations, a parasitology reference laboratory with a high incidence of immunocompromised patients and a university research center, to validate SUBSTANTIAL EQUIVALENCE of the device.

Fecal samples known to be positive or negative for *Cryptosporidium parvum* by conventional microscopy with modified acid fast (MAF) staining were tested. Sensitivity / Specificity values were calculated and compared with the Sensitivity/ Specificity values for the referenced predicate device. Substantial equivalency was validated by the testing.

## Reference Data for Predicate Kit:

| Study #1: | Study #2: (Resolved) |
| --- | --- |
| Sensitivity: 97% | Sensitivity: 97% |
| Specificity: 100% | Specificity: 98% |
| Microscopy + = 134 | Microscopy + = 35 |
| Microscopy - = 78 | Microscopy - = 343 |
| Study Base: 212 | Study Base: 378 |

## TREND Cryptosporidium Performance Data:

| Study A: | Study B: |
| --- | --- |
| Sensitivity: 96.2% | Sensitivity: 97% |
| Specificity: 97.1% | Specificity: 100% |
| Microscopy + = 26 | Microscopy + = 68 |
| Microscopy - = 70 | Microscopy - = 29 |
| Study Base: 96 | Study Base: 97 |

0496

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Page 4 of 4

Summary of 510(k) Safety and Effectiveness information.
Trade / Proprietary Name: TREND Cryptosporidium Direct Detection
Test System, Cat. # EA49-

Additional Testing:

Further bench studies validated performance equivalency in the following parameters:

- Parallel studies were performed using referenced specimen preparations and "types".
- Testing of known parasite positive fecal specimens was performed to validate specificity/ freedom from cross reactivity. No cross reactivity was seen with any of the parasitic samples tested. Following is a list of the tested parasitic organisms:

Ascaris lumbricoides
Cyclospora spp.
Entamoeba coli
Endolimax nana
Hymenolepis nana
Paragonimus wester.
Trichomonas hominis
Blastocystis hominis
Dientamoeba fragilis
Entamoeba hartmanni
Fasciola hepatica
Iodamoeba butschlii
Strongyloides sterc.
Hookworm
Chilomastix mesnili
Diphyllobothrium latum
Entamoeba histolytica
Giardia lamblia
Isospora belli
Taenia spp.
Clonorchis sinensis
Eimeria spp.
Entamoeba polecki
Hymenolepis diminuta
Microsporidia spp.
Trichuris trichuria

No cross reactivity was seen with any of the following fecal source microorganisms:

Pathogenic Enteric Bacteria:
Campylobacter jejuni
Escherichia coli
Salmonella typhimurium
Shigella flexneri
Fungi:
Candida albicans
Cryptococcus neoformans
Virus:
Rotavirus

- Testing to determine Analytical Sensitivity of the Test System was performed. The sensitivity was determined to be approximately 4.6 ng/ml. of Cryptosporidium antigen.
- Testing for Precision and Reproducibility was performed at three off-site locations.

SUBSTANTIALLY EQUIVALENCE CONCLUSIONS:

The above provided information validates that the TREND device is substantially equivalent to the referenced predicate kit.

- The Intended Use of the kit is the same.
- Scientific Principle / Technology is the same.
- Performance Data, as presented, validates that the performance (Sensitivity / Specificity) is equivalent.
- Analytical Sensitivity of the device is equivalent.

#496

Prepared by: Mariellyn Truax, MT (ASCP)
Title: Director of Quality Control
Date: 4/24/96