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

Device Description

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.

AI/ML Overview

The TREND Cryptosporidium Direct Detection Test System is an ELISA test kit intended for the qualitative determination of Cryptosporidium antigen in feces from patients with diarrhea to aid in the detection of Cryptosporidium gastrointestinal infection. The study aimed to validate the substantial equivalence of the device to a referenced predicate device.

1. Table of Acceptance Criteria and Reported Device Performance:

The document implicitly defines acceptance criteria through comparison to a predicate device. Substantial equivalence is validated if the performance (sensitivity/specificity) is equivalent.

Metric	Predicate Device Study 1	Predicate Device Study 2 (Resolved)	TREND Device Study A	TREND Device Study B	Acceptance Criteria (Implied by Predicate)
Sensitivity	97%	97%	96.2%	97%	Sensitivity ≥ 97% (based on Study 1 & 2) or comparable
Specificity	100%	98%	97.1%	100%	Specificity ≥ 98% (based on Study 2) or comparable

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

The test set consisted of fecal samples known to be positive or negative for Cryptosporidium parvum by conventional microscopy with modified acid-fast (MAF) staining.

Study A: 96 samples (26 Microscopy +, 70 Microscopy -)
Study B: 97 samples (68 Microscopy +, 29 Microscopy -)

The data provenance is stated as "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." This suggests a combination of retrospective (known positive/negative samples) and potentially prospective (samples from the high incidence lab) data, although it's not explicitly detailed as retrospective or prospective. The country of origin is not specified.

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

The ground truth was established by "conventional microscopy with modified acid-fast (MAF) staining." The document does not specify the number of experts who performed these microscopic examinations or their qualifications (e.g., years of experience as a microbiologist or parasitologist).

4. Adjudication Method for the Test Set:

The document doesn't explicitly describe an adjudication method for the ground truth. It simply states that samples were "known to be positive or negative for Cryptosporidium parvum by conventional microscopy with modified acid fast (MAF) staining." This implies a single determination or a standard laboratory process was used without a separate adjudication panel.

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

No, an MRMC comparative effectiveness study was not done. The study compares the performance of the device to a predicate device and standard microscopy. It does not measure the improvement of human readers with AI assistance.

6. If a Standalone (Algorithm Only Without Human-in-the-Loop Performance) Was Done:

Yes, a standalone study was performed. The performance data presented (Sensitivity and Specificity for Study A and Study B) relate directly to the output of the ELISA test kit (the "algorithm" in this context) without a human-in-the-loop component influencing the primary test result. The results are read spectrophotometrically or visually, but the core performance metrics are for the test system itself.

7. The Type of Ground Truth Used:

The primary ground truth used was expert consensus via conventional microscopy with modified acid-fast (MAF) staining. This is a laboratory-based diagnostic method.

8. The Sample Size for the Training Set:

The document does not provide information about a separate training set. The study describes performance testing using clinical laboratory bench studies. For this type of ELISA kit, robust training sets in the AI sense are typically not discussed, as the "training" (calibration, optimization) of the assay components and parameters would be part of the manufacturing and development process, rather than a distinct data-driven training phase after the assay design is fixed for performance testing.

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

Since a training set (in the AI context) is not explicitly mentioned, information on how its ground truth was established is not provided. The development and optimization of such assays would generally involve internal validation against well-characterized samples, but this is distinct from the formal performance testing described.

Summary

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Summary of 510(k) Safety and Bffectiveness information. Trade / Proprietary Name: TRBND Cryptosporidium Direct Detection
Test System, Cat.4 BA49-

DESCRIPTION and INTENDED USE of the DEVICE:

Traditional detection of parasitic organisms has been conventional microscopic examination of fecal material. During the last few vears. 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 selflimiting 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:

TECHNOLOGY:

ﺳﯩﻤﻪ

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. છે 4 તે ર

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Summary of 510(k) Safety and Bffectiveness information. TREAD Cryptosporidium Direct Detection Trade / Proprietary Mane:
Test System, Cat. # BA49-

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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Summary of 510(k) Safety and Effectiveness information. Page 4 of 4 TREND Cryptosporidium Direct Detection Trade / Proprietary Wame: Test System, Cat.4 BA49-Additional Testing: Further bench studies validated performance equivalency in the

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 lumbricoidesCyclospora spp.Entamoeba coliEndolimax nanaHymenolepis nanaParagonimus westeri.Trichomonis hominis	Blastocystis hominisDientamoeba fragilisEntamoeba hartmanniFasciola hepaticaIodamoeba butschliiStrongyloides sterc.Hookworm	Chilomastix mesniliDiphyllobothrium latumEntamoeba histolyticaGiardia lambliaIsospora belliTaenia spp.	Clonorchis sinensisEimeria spp.Entamoeba poleckiHymenolepis diminutaMicrosporidia spp.Trichuris trichiura
------------------------------------------------------------------------------------------------------------------------------------------------	-----------------------------------------------------------------------------------------------------------------------------------------------------	----------------------------------------------------------------------------------------------------------------------------	-------------------------------------------------------------------------------------------------------------------------------

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

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

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

SUBSTANTIALLY EQUIVALENCY CONCLUSIONS:

following parameters:

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.

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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.