The TECHLAB® TRI-COMBO PARASITE SCREEN test is an enzyme immunoassay for the simultaneous qualitative detection of Giardia spp., Cryptosporidium spp. and/or E. histolytica antigen in human fecal specimens. The test is indicated as an aid in the diagnosis of gastrointestinal infection when giardiasis, cryptosporidiosis and amebiasis is suspected. The test does not differentiate between the three parasites and follow-up testing is required for all positive results to confirm the specific diagnosis.

Device Description

The TRI-COMBO PARASITE SCREEN test is an enzyme immunoassay for the simultaneous qualitative detection of Giardia spp., Cryptosporidium spp. and/or E. histolytica antigen in human fecal specimens. The test uses monoclonal and polyclonal antibodies to cell-surface antigens of Giardia, Cryptosporidium and E. histolytica. The microassay plate in the kit contains immobilized monoclonal antibodies against the antigens, and the Conjugate consists of polyclonal antibodies against the antigens. In the assay, an aliquot of a diluted fecal specimen is transferred to a microassay well. The immobilized monoclonal antibodies bind the Giardia, Cryptosporidium and/or E. histolytica antigens if they are present. Upon addition, Conjugate then binds to the antigen/ antibody complex. Any unbound materials are removed during the washing steps. Following the addition of Substrate, a color is detected due to the enzyme-antibody-antigen complexes that formed in the presence of antigens and conjugate.

AI/ML Overview

Here's an analysis of the provided text regarding the TECHLAB® TRI-COMBO PARASITE SCREEN test, structured according to your requested points:

1. A table of acceptance criteria and the reported device performance

The document does not explicitly state pre-defined acceptance criteria (e.g., "The device must achieve sensitivity of X% and specificity of Y%"). Instead, it presents performance data which implicitly serves as the basis for the FDA's substantial equivalence decision. Therefore, I will present the reported performance data from the prospective clinical study as the proxy for "acceptance criteria met." For the retrospective study, while providing higher values, the prospective study is generally weighted more heavily for regulatory approval as it reflects real-world conditions more closely. The document also provides "95% Confidence Limits" for sensitivity and specificity.

Metric (Prospective Study vs. Microscopy)	Reported Device Performance	95% Confidence Limits
Sensitivity	92.9%	68.5% - 98.7%
Specificity	98.1%	96.9% - 98.9%
Positive Percent Agreement (vs. Microscopy and Molecular)	62.1%	44.0% - 77.3%
Negative Percent Agreement (vs. Microscopy and Molecular)	98.8%	97.7% - 99.4%

2. Sample size used for the test set and the data provenance (e.g., country of origin of the data, retrospective or prospective)

Prospective Study:
- Sample Size: 754 (740 negative, 14 microscopy positive - 13 Giardia, 1 E. histolytica)
- Data Provenance: Conducted at 3 independent clinical sites. Specific country of origin is not explicitly stated, but the submission is to the U.S. FDA by a U.S. company, suggesting U.S. clinical sites. The data is prospective.
Retrospective Study:
- Sample Size: 96 archived specimens (previously collected and frozen)
- Data Provenance: From one clinical site in an E. histolytica endemic area (not specified by country). The data is retrospective.

3. Number of experts used to establish the ground truth for the test set and the qualifications of those experts (e.g., radiologist with 10 years of experience)

The document primarily refers to "microscopy" and "molecular testing."

Microscopy: It states that "accuracy of O&P results depends upon the skill of the technician." This implies trained laboratory technicians or medical technologists are performing microscopy. However, the exact number of experts or their specific qualifications (e.g., board certification, years of experience) are not provided in the document.
Molecular Testing: The document mentions "molecular testing of a commercial FDA cleared device and PCR with sequencing." It does not specify the number or qualifications of experts involved in interpreting or performing these molecular tests.

4. Adjudication method (e.g., 2+1, 3+1, none) for the test set

The document does not describe a formal adjudication method (e.g., 2+1, 3+1) involving multiple human readers for discrepancies. Instead, discrepancies between the device and microscopy were resolved via further testing:

For the prospective study, the 14 TRI-COMBO PARASITE SCREEN positives that were microscopy negative were "confirmed to be positive for Giardia with an alternate FDA cleared antigen test or by PCR with sequencing." This constitutes a form of resolution by a more definitive method rather than human adjudication.

5. 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

This is not an AI-assisted device for human readers. The device is an in vitro diagnostic (IVD) immunoassay for detecting antigens in fecal specimens. Therefore, an MRMC comparative effectiveness study involving human readers improving with AI assistance is not applicable to this device. The comparison is between the immunoassay and traditional methods (microscopy and molecular testing).

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

Yes, the device is a standalone diagnostic test (an immunoassay). Its performance is evaluated directly against comparator methods (microscopy, PCR, other FDA-cleared antigen tests) without human interpretation of the device's generated data being a variable. The test results are "spectrophotometric" or "visual" interpretations of a color change, but the core performance data reflects the device's ability to detect the analytes by itself.

7. The type of ground truth used (expert consensus, pathology, outcomes data, etc)

The ground truth for the test sets was established using a combination of:

Microscopy: Primarily light microscopy for identification of Giardia, Cryptosporidium, and E. histolytica.
Molecular Testing:
- For the prospective study, "composite results from multiple tests that consisted of light microscopy, molecular testing of a commercial FDA cleared device and PCR with sequencing for the identification of Giardia spp., Cryptosporidium spp., in addition to identification and subspeciation of E. histolytica." This "composite" is considered the ground truth.
- For discrepancies, "alternate FDA cleared antigen test or by PCR with sequencing" was used.
Archived Specimen Characterization: For the retrospective study, specimens were characterized as "microscopy and PCR positive."

8. The sample size for the training set

The document describes performance studies for the device but does not mention a training set because this is an in vitro diagnostic (IVD) immunoassay, not a machine learning or AI-based device that requires a training set. The design of the assay (antibodies, reagents) is based on scientific principles and previous research, not on learning from a specific dataset.

9. How the ground truth for the training set was established

As there is no training set for this type of device, this question is not applicable.

Summary

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Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002

July 10, 2017

TECHLAB, INC. DONNA LINK DIRECTOR REGULATORY AND COMPLIANCE 2001 KRAFT DRIVE BLACKSBURG VA 24060-6358

Re: K171078

Trade/Device Name: Tri-combo Parasite Screen Regulation Number: 21 CFR 866.3220 Regulation Name: Entamoeba histolyticaserological reagents Regulatory Class: II Product Code: MHJ, MHI and KHW Dated: April 10, 2017 Received: April 11, 2017

Dear Ms. Link:

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, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21. Parts 800 to 898. 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); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

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If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638 2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to

http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.

You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely yours,

Ribhi Shawar -S For

Uwe Scherf, M.Sc., Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

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Indications for Use

510(k) Number (if known) K171078

Device Name

TRI-COMBO PARASITE SCREEN

Indications for Use (Describe)

The TECHLAB® TRI-COMBO PARASITE SCREEN test is an enzyme immunoassay for the simultaneous qualitative detection of Giardia spp., Cryptosporidium spp. and/or E. histolytica antigen in human fecal specimens. The test is indicated as an aid in the diagnosis of gastrointestion when giardiasis, cryptosporidiosis is suspected. The test does not differentiate between the three parasites and follow-up testing is required for all positive results to confirm the specific diagnosis.

Type of Use (Select one or both, as applicable)
X Prescription Use (Part 21 CFR 801 Subpart D)	Over-The-Counter Use (21 CFR 801 Subpart C)

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TRI-COMBO PARASITE SCREEN 510(k) SUMMARY

This summary of 510(k) safety and effectiveness is being submitted in accordance with the requirements of 21 CFR 807.92.

Applicant/Contact Information:

Date Prepared:	July 10, 2017
Name:	TECHLAB, Inc.
Address:	2001 Kraft DriveCorporate Research CenterBlacksburg, VA 24060 USA
Contact Person:	Donna T. Link
Phone Number:	540-953-1664
Email:	dlink@techlab.com

1.1 Manufacturing Facility Address

TECHLAB, Inc. 20 Corporate Drive Radford, VA 24141 USA

1.2 Product and Trade Name of the Device

TRI-COMBO PARASITE SCREEN

1.3 Common Name or Classification Name

Giardia spp., Cryptosporidium spp., and E. histolytica detection test

1.4 Classification and Regulation

Class II

21 CFR 866.3220; Entamoeba histolytica serological reagents

1.5 Product Code

MHJ - Cryptosporidium spp.

1.6 Panel

83 Microbiology

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Intended Use

The TECHLAB® TRI-COMBO PARASITE SCREEN test is an enzyme immunoassay for the simultaneous qualitative detection of Giardia spp., Cryptosporidium spp. and/or E. histolytica antigen in human fecal specimens. The test is indicated as an aid in the diagnosis of gastrointestinal infection when qiardiasis, cryptosporidiosis and amebiasis is suspected. The test does not differentiate between the three parasites and follow-up testing is required for all positive results to confirm the specific diagnosis.

Explanation

The most common method used to detect giardiasis, cryptosporidiosis and amebiasis has been diagnosis by ova and parasite (O&P) light microscopy. However, accuracy of O&P results depends upon the skill of the technician and also relies on the presence of intact cysts in the feces, which may not be present in all samples. In addition, it is only rarely possible to distinguish between the pathogenic and non-pathogenic species of Entamoeba using microscopy. The success rate of fecal examination by microscopy varies between 50 and 70%, and multiple specimens are usually necessary to establish a diagnosis. When infection is present but parasites are not detected by microscopy, sampling and testing of duodenal fluid may detect trophozoites, but this method is invasive and expensive. Detection of the organism and antigens by ELISA provides an alternative method of diagnosis and is sensitive and specific. The TRI-COMBO PARASITE SCREEN ELISA procedure is straightforward to perform and exhibits increased sensitivity compared to microscopic examination. Large numbers of specimens can be tested rapidly and objectively, and the procedure is less labor-intensive than most microscopy methods.

Device Description

Materials Provided

. Conjugate (7 mL) – Antibodies against antigens of Giardia spp., Cryptosporidium spp., and E. histolytica in a buffered protein solution containing 0.05% ProClin® 300
. Diluent (50 mL) – Buffered protein solution containing 0.02% thimerosal. The Diluent is also to be used as the negative control solution
. Stop Solution (7 mL) – 0.6 N sulfuric acid. Caution: Avoid contact with skin: flush with water immediately if contact occurs
. Giardia Positive Control (3.5 mL) – Giardia antigen in a buffered protein solution with 0.02% thimerosal
Cryptosporidium Positive Control (3.5 mL) Cryptosporidium antigen in a buffered protein . solution with 0.02% thimerosal
. E. histolytica Positive Control (3.5 mL) – E. histolytica antigen in a buffered protein solution with 0.02% thimerosal

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Substrate (14 mL) – solution containing tetramethylbenzidine and peroxide
Wash Buffer Concentrate (50 mL) 20X concentrate containing phosphate buffered saline, . detergent, and 0.2% thimerosal
. Microassay Plate - 12 strips, each consisting of 8 wells coated with antibodies to Giardia spp., Cryptosporidium spp., and E. histolytica antigen (stored with desiccant)
Disposable plastic transfer pipettes - Quantity 100
. Plastic adhesive sheets - Quantity 2 Sheets
Wash Solution Label - Quantity 1

Predicate Device Information

The predicate device (GIARDIA/CRYPTOSPORIDIUM CHEK®) and the TRI-COMBO PARASITE SCREEN test use the same EIA (enzyme immunoassay) technology and are substantially equivalent in principle. The following tables show a comparison of both devices' similarities and differences.

Similarities
ltem	Device (K171028)	Predicate (K051929)
Intended Use	The TECHLAB® TRI-COMBO PARASITESCREEN test is an enzyme immunoassay forthe simultaneous qualitative detection ofGiardia spp., Cryptosporidium spp. and/or E.histolytica antigen in human fecal specimens.The test is indicated as an aid in the diagnosisof gastrointestinal infection when giardiasis,cryptosporidiosis and amebiasis is suspected.The test does not differentiate between thethree parasites and follow-up testing isrequired for all positive results to confirm thespecific diagnosis.	The GIARDIA/CRYPTOSPORIDIUM CHEK test isan enzyme immunoassay for thequalitative detection of Giardia cystand Cryptosporidium oocyst antigenin human fecal specimens. It isindicated for use as an aid in thediagnosis of patients with diarrheasuspected of Giardia and/orCryptosporidium gastrointestinalinfections
Technology	Enzyme Linked Immunoassay (ELISA)	Same
Antibody Format	Monoclonal capture Ab Polyclonal secondaryAb	Same
Type of Test	Qualitative	Same
Format/Tests	Microassay Well Plate (96 tests)	Same
Controls	Positive and negative control are included inthe kit	Same
Interpretation	Spectrophotometrically and visually	Same

Differences
Item	Device	Predicate
Analyte Detected	Giardia spp., Cryptosporidiumspp., and E. histolytica specificantigens	Giardia spp., and Cryptosporidium spp.,specific antigens
Acceptable Specimen Type	Fecal specimens in Cary-Blairand C&S Transport Media	Specimens in preservation media of 10%buffered formalin or Sodium AcetateFormalin (SAF)

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Summary of Performance Data

Prospective Study

The performance of the TRI-COMBO PARASITE SCREEN test was evaluated at 3 independent sites. The three sites yield a total of 14 microscopy positive samples (13 Giardia and one E. histolytica). The remaining 740 samples were negative. Of the 740 negative samples, four were E. histolytica positive by molecular comparison only but were negative by the TRI-COMBO PARASITE SCREEN test, negative by microscopy, and negative by FDA cleared antigen test for E. histolytica. Table 1 summarizes the performance observed, which is primarily a study to evaluate specificity due to the low number of positive specimens (see retrospective study for evaluation of sensitivity). Prospective testing was also read visually and performance was not significantly different from spectrophotometric readings.

Table 1. Summary of prospective clinical performance comparing the TRI-COMBO PARASITE SCREEN test to microscopy for Giardia. Cryptosporidium and E. histolytica

TRI-COMBO PARASITE SCREEN(N = 754)	Microscopy
	Positive	Negative
Positive	13	14*
Negative	1	726
		95% Confidence Limits
Sensitivity	92.9%	68.5% - 98.7%
Specificity	98.1%	96.9% - 98.9%

Specimens Positive for Giardia by microscopy/Specimens Detected by the TRI-COMBO PARASITE SCREEN	12/13
Specimens Positive for E. histolytica by microscopy/Specimens Detected by the TRI-COMBO PARASITE SCREEN	1/1

*The fourteen TRI-COMBO PARASITE SCREEN positives that were microscopy negative were confirmed to be positive for Giardia with an alternate FDA cleared antigen test or by PCR with sequencing.

Retrospective Study

Testing consisted of 96 archived specimens previously collected and frozen from one clinical site. The Frozen samples are included in the bank based on being characterized as microscopy and PCR positive. The specimens were collected from an E. histolytica endemic area and contained specimens also positives for Giardia and Cryptosporidium. Table 2 summarizes the performance observed. Retrospective testing was also read visually and performance was not different from spectrophotometric readings.

Table 2. Summary of retrospective clinical performance comparing the TRI-COMBO PARASITE SCREEN test to Microscopy and PCR

TRI-COMBO PARASITE SCREEN	Microscopy and PCR
(N = 96)	Positive	Negative
Positive	85	0
Negative	5	6

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		95% Confidence Limits
Sensitivity	94.4%	87.7% - 97.9%
Specificity	100%	61.0% - 100%
Specimens Positive for Giardia/	Specimens Detected by the TRI-COMBO PARASITE SCREEN	41/41
Specimens Positive for Cryptosporidium/	Specimens Detected by the TRI-COMBO PARASITE SCREEN	27/30
Specimens Positive for E. histolytica/	Specimens Detected by the TRI-COMBO PARASITE SCREEN	28/30
Note: Eight specimens were dual positive for Giardia and E. histolytica by the Microscopy and PCR and
tested positive in the TRI-COMBO PARASITE SCREEN test. Three specimens were dual positive for
Giardia and Cryptosporidium by Microscopy and PCR and tested positive in the TRI-COMBO PARASITE
SCREEN test.

The prospective study results were analyzed by considering composite results from multiple tests that consisted of light microscopy, molecular testing of a commercial FDA cleared device and PCR with sequencing for the identification of Giardia spp., Cryptosporidium spp., in addition to identification and subspeciation of E. histolytica. This testing was mainly done because identification of E. histolytica organisms cannot be determined solely by microscopy because it is morphologically indistinguishable from the non-pathogenic E. dispar. Use of an alternate molecular testing is needed to confirm Entamoeba speciation. The molecular testing algorithm used provides a comparator method that is highly sensitive at the detection of Giardia spp., Cryptosporidium spp. and E. histolytica. The performance is summarized in table 3 and is presented as positive percent agreement and negative percent agreement.

Table 3. Summary of prospective clinical performance comparing the TRI-COMBO PARASITE SCREEN test to microscopy and molecular testing

TRI-COMBO PARASITE SCREEN(N = 754)	Microscopy and Molecular testing
	Positive	Negative
Positive	18	9*
Negative	11**	716
Positive Percent Agreement	62.1%	44.0% - 77.3%
Negative Percent Agreement	98.8%	97.7% - 99.4%

Specimens Positive for Giardia/Specimens Detected by the TRI-COMBO PARASITE SCREEN	17/24
Specimens Positive for E. histolytica/Specimens Detected by the TRI-COMBO PARASITE SCREEN	1/5
Specimens Positive for Cryptosporidium/Specimens Detected by the TRI-COMBO PARASITE SCREEN	0/0

' These nine specimens were tested with an alternate FDA cleared antigen test resulting in 99 Giardia determined to be antigen positive.

**These eleven specimens were tested with an alternate FDA cleared antigen test resulting in 6/7 Giardia and 4/4 E. histolytica were determined to be antigen negative.

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Reproducibility

The reproducibility of the TRI-COMBO PARASITE SCREEN test was determined using 20 human fecal specimens coded to prevent their identification during testing. Testing was performed at 2 independent laboratories and on-site at TECHLAB, Inc. The samples were tested twice a day over a 5-day period by multiple technicians at each site using 2 different kit lots. Positive and negative controls were run with each panel of the masked samples. The results from each laboratory were submitted to TECHLAB, Inc. and compared with in-house results. The results were consistent among the different locations and exhibited a correlation of 99.9%. The samples produced the expected results 99.9% of the time.

Analytical Sensitivity

Limit of Detection (LoD) – cutoff points for Giardia cysts, Cryptosporidium oocysts and E. histolytica pathogenic zymodemes in fecal specimens for the TRI-COMBO PARASITE SCREEN test. Determined following CLSI document EP17-A2.

CLSI. Evaluation of Detection Capability for Clinical Laboratory Measurement Procedures: Aproved Guideline - Second Edition. CLSI document EP17-A2. Wayne, PA: Clinical and Laboratory Standards Institute; 2012.

The analytical sensitivity of the test was determined by using purified Giardia cysts, Cryptosporidium oocysts, and E. histolytica pathogenic zymodemes in a sample matrix. The concentration of Giardia cysts, Cryptosporidium oocysts, and E. histolytica pathogenic zymodemes in fecal matrix of which specimens were positive by the TRI-COMBO PARASITE SCREEN test 95% of the time was the assay limit-of-detection (LoD). Test results determined the LoD for the assay to be 8450 cysts/mL of feces for Giardia (equivalent to 169 cysts detected per test), 47,962 oocysts/mL of feces for Cryptosporidium (equivalent to 959 oocysts detected per test), and 1.676 PZs/mL (equivalent to 34 PZs detected per test). For fecal matrix/Cary Blair, the LoD for the assay was determined to be 34,155 cysts/mL of feces for Giardia (equivalent to 427 cysts detected per test), 99,456 oocysts/mL of feces for Cryptosporidium (equivalent to 1243 occysts detected per test), and 4655 PZs/mL (equivalent to 58 PZs detected per test). For fecal matrix/C&S, the LoD for the assay was determined to be 37.095 cysts/mL of feces for Giardia (equivalent to 464 cysts detected per test), 122,299 occysts/mL of feces for Cryptosporidium (equivalent to 1529 ocysts detected per test), and 3948 PZs/mL (equivalent to 49 PZs detected per test).

Because the TRI-COMBO PARASITE SCREEN test detects soluble antigen in fecal specimens in addition to cysts, oocysts, and trophozoites, this LOD study represents an estimate of analytical sensitivity.

Analytical Specificity (Cross Reactivity)

The TRI-COMBO PARASITE SCREEN test was evaluated for cross-reactivity with the bacterial and viral strains listed below. None of the strains were shown to interfere with the performance of the TRI-COMBO PARASITE SCREEN test.

Aeromonas hydrophila Bacteroides fragilis Campylobacter jejuni Clostridium difficile Escherichia coli 0157:H7 Escherichia coli ETEC Shigella dysenteriae Staphylococcus aureus Vibrio parahaemolyticus

Bacillus cereus Campylobacter coli Candida albicans Enterococcus faecalis Escherichia coli EIEC Klebsiella pneumonia Shigella flexneri Staphylococcus aureus (Cowan's) Yersinia enterocolitica

Bacillus subtilis Campylobacter fetus Clostridium bifermentans Escherichia coli Escherichia coli EPEC Salmonella typhimurium Shigella sonnei Staphylococcus epidermidis

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Cytomegalovirus Echovirus 11. 18. 33 Calicivirus Human Adenovirus 1, 2, 3, 5, 40, 41 Human Coronavirus Human Coxsackievirus B2, B3, B4, B5 Human Echovirus 9 Human Enterovirus 68, 69, 70, 71 Human parechovirus 1 [Echovirus 22] Human Rotavirus

Additionally, the TRI-COMBO PARASITE SCREEN test was run on fecal specimens documented to be positive for other parasites by microscopy. The number in parentheses is the quantity of each organism found in the clinical specimens. No cross-reactivity was seen with the following organisms.

Ascaris lumbricoides and with eggs (22)	Entamoeba coli (16)	Trichuris trichiura eggs (12)
Blastocystis hominis (11)	Entamoeba moshkovskii (3)
Entamoeba bangladeshi (3)	Iodamoeba bütschlii (10)

Cross reactivity with Norovirus is unknown because it was not tested in analytical studies. However, Norovirus GI/GII was identified in 34 clinical specimens and Enterotoxigenic E. Coli - ETEC LT/ST was identified in 107 clinical specimens, using an FDA cleared multiplex NAAT assay during clinical testing and no cross reactivity was found using the TRI-COMBO PARASITE SCREEN in those samples.

Strain Specific Study

Due to the similarity in morphology between pathogenic and non-pathogenic Entamoeba species, 3 specimens identified by PCR as positive for non-pathogenic Entamoeba moshkovskii and 3 positive for non-pathogenic Entamoeba bangladeshi were evaluated using the TRI-COMBO PARASITE SCREEN test. These 6 specimens tested negative in the TRI-COMBO PARASITE SCREEN test.

Interfering Substances (U.S. Formulation)

The following substances had no effect on positive or negative TRI-COMBO PARASITE SCREEN test results analyzed at the concentrations indicated:

Barium sulfate (5% w/v), Benzalkonium Chloride (1% w/v), Ciprofloxacin (0.25% w/v), Ethanol (1% w/v), Hog gastric mucin (3.5% w/v), Human blood (40% v/v), Hydrocortisone (1% w/v), Imodium® (5% v/v), Kaopectate® (5% v/v), Leukocytes (0.05% w/v), Maalox® Advanced (5% v/v), Mesalazine (10% w/v), Metronidazole (0.25% w/v), Mineral Oil (10% w/v), Mylanta® (4.2 mg/mL), Naproxen Sodium (5% w/v), Nonoxynol-9 (1% w/v), Nystatin (1% w/v), Palmitic Acid/Fecal Fat (40% w/v), Pepto-Bismol® (5% v/v), Phenylephrine (1% w/v), Polyethylene glycol 3350 (10% w/v ), Prilosec OTC® (5 µg/mL), Sennosides (1% w/v), Simethicone (10% w/v), Stearic Acid/Fecal Fat (40% w/v), Tagamet® (5 µg/mL), TUMS (50 µg/mL), Human Urine (5% v/v), and Vancomycin (0.25% w/v).

Precision - Intra-assay

For the determination of intra-assay performance. 24 fecal samples were analyzed by the TRI-COMBO PARASITE SCREEN test. The samples included 2 negative, 2 high negative, 2 low positive, and 2 moderate positive samples for each analyte. Each specimen was assayed a total of five times using two different kit lots. Positive specimens consistently tested positive and negative specimens consistently tested negative. High negative samples tested within 95% agreement of each other for all three analytes.

Precision - Inter-assay

For the determination of inter-assay performance, 24 fecal samples were analyzed by the TRI-COMBO PARASITE SCREEN test. The samples included 2 negative, 2 high negative, 2 low positive, and 2 moderate positive samples for each analyte. The samples were tested twice a day by multiple

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technicians over a 12-day period using 2 different kit lots. All positive samples remained positive and all negative samples remained negative.

Fresh Versus Frozen Samples

The effect of long term frozen specimen storage on antigen stability was evaluated. For the analysis, a total of 39 fecal samples were tested with the TRI-COMBO PARASITE SCREEN test. The samples consisted of negative fecal samples, high negative fecal samples, low positive fecal samples, moderate positive fecal samples, and high positive fecal samples. Samples were prepared by spiking a negative fecal pool with Giardia cysts, Cryptosporidium oocysts, or E. histolytica pathogenic zymodemes at respective concentrations and stored at ≤ -10°C. These samples were tested at 0, 1, 4 and 8 weeks. No conversion of positive-to-negative-to-positive was observed in any of the samples at the specified time points.

Prozone

To ensure that a high concentration of analyte does not interfere with a positive reaction in TRI-COMBO PARASITE SCREEN test, high samples were prepared by spiking a negative fecal pool with Giardia cysts, Cryptosporidium oocysts, or E. histolytica pathogenic zymodemes, and then tested. A total of 5 different dilutions of each, up to and including the clinically observed high concentration, were prepared and tested in triplicate. The results demonstrated that there was no overall prozone affect, that elevated levels of analyte did not affect the detection of each organism.

Conclusion

The conclusions drawn from the nonclinical and clinical tests demonstrate that the TRI-COMBO PARASITE SCREEN test is safe and effective for the detection of Giardia spp., Cryptosporidium spp., and E. histolytica in human fecal specimens. The information submitted in this premarket notification is complete and supports a substantial equivalence decision.

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.