GIARDIA/ CRYPTOSPORIDIUM QUIK CHEK

{0} 1 # 510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION DECISION SUMMARY ASSAY ONLY TEMPLATE A. 510(k) Number: K103673 B. Purpose for Submission: To obtain a substantial equivalence determination for the detection of Giardia and Cryptosporidium antigens in human stool C. Measurand: Giardia and Cryptosporidium antigens D. Type of Test: Qualitative membrane enzyme immunoassay E. Applicant: TechLab® Inc. F. Proprietary and Established Names: The GIARDIA/CRYPTOSPORIDIUM QUIK CHEK™ test G. Regulatory Information: 1. Regulation section: 21 CFR Part 866.3220 Entamoeba histolytica Serological Reagents 2. Classification: II 3. Product code: MHJ - Cryptosporidium spp. MHI - Giardia spp. {1} 4. Panel: 83, Microbiology H. Intended Use: 1. Intended use(s): The GIARDIA/CRYPTOSPORIDIUM QUIK CHEK test is a rapid membrane enzyme immunoassay for the simultaneous qualitative detection and differentiation of Giardia cyst antigen and Cryptosporidium oocyst antigen in a single test device. It is intended for use with human fecal specimens from patients with gastrointestinal symptoms to aid in the diagnosis of Giardia and/or Cryptosporidium gastrointestinal infection. As with other Giardia and/or Cryptosporidium tests, results should be considered in conjunction with the patient history. FOR IN VITRO DIAGNOSTIC USE. 2. Indication(s) for use: The GIARDIA/CRYPTOSPORIDIUM QUIK CHEK test is a rapid membrane enzyme immunoassay for the simultaneous qualitative detection and differentiation of Giardia cyst antigen and Cryptosporidium oocyst antigen in a single test device. It is intended for use with human fecal specimens from patients with gastrointestinal symptoms to aid in the diagnosis of Giardia and/or Cryptosporidium gastrointestinal infection. As with other Giardia and/or Cryptosporidium tests, results should be considered in conjunction with the patient history. 3. Special conditions for use statement(s): For prescription use only 4. Special instrument requirements: None I. Device Description: The GIARDIA/CRYPTOSPORIDIUM QUIK CHEK™ test device contains 25 pouches, each containing a membrane filter pad. Each device has a Reaction Window with three vertical lines of immobilized antibodies. The Giardia test line ("Giar") contains mouse monoclonal antibodies against Giardia. The Crypto test line ("Cryp") contains mouse monoclonal antibodies against Cryptosporidium. The control line ("C") is a dotted line that contains anti-horseradish peroxidase (HRP) antibodies. The 2 {2} Conjugate consists of polyclonal antibodies coupled to horseradish peroxidase. The antigen-antibody-conjugate complexes migrate through a filter pad to a membrane where they are captured by the immobilized Giardia and/or Cryptosporidium-specific antibodies in the test lines. After addition of substrate and proper incubation, the reaction is examined visually for the appearance of a vertical blue line on either side of the Reaction Window. A blue line indicates a positive test. A positive "control" reaction, indicated by a vertical dotted blue line under the "C" portion of the Reaction Window, confirms that the test is working properly and the results are valid. ## J. Substantial Equivalence Information: 1. Predicate device name(s): GIARDIA II CRYPTOSPORIDIUM II 2. Predicate 510(k) number(s): K033274 K052932 3. Comparison with predicate: | Similarities | | | | | --- | --- | --- | --- | | Item | Device | Predicate 1 | Predicate 2 | | Intended Use | Detection of Giardia cyst and Cryptosporidium oocyst antigen | Detection of Giardia cyst antigen | Detection of Cryptosporidium oocyst antigen | | Technology | Enzyme Immunoassay | Enzyme Immunoassay | Enzyme Immunoassay | | Antibody Format | Monoclonal/Polyclonal | Monoclonal/Polyclonal | Monoclonal/Polyclonal | | Specimen Types | Human Fecal Specimens- fresh, frozen, or preserved in 10% Formalin or SAF | Human Fecal Specimens- fresh, frozen, or preserved in 10% Formalin or SAF | Human Fecal Specimens- fresh, frozen, or preserved in 10% Formalin or SAF | {3} | Differences | | | | | --- | --- | --- | --- | | Item | Device | Predicate 1 | Predicate 2 | | Intended Use | Detection and differentiation of *Giardia* cyst antigen and *Cryptosporidium* oocyst antigen | Detection *Giardia* cyst antigen only | Detection *Cryptosporidium* oocyst antigen only | | Technology | Membrane Enzyme Immunoassay | Enzyme Immunoassay – Microassay Plate ELISA | Enzyme Immunoassay – Microassay Plate ELISA | | Amount of Specimen required | 25 μL – fresh 100 μL - preserved | 100 μL – fresh or preserved | 100 μL – fresh or preserved | | Incubation time | 15 minutes | 1 hour | 1 hour | | Time to Result | 30 minutes | 1 hour 45 minutes | 1 hour 45 minutes | ## K. Standard/Guidance Document Referenced (if applicable): Not applicable ## L. Test Principle: The GIARDIA/CRYPTOSPORIDIUM QUIK CHEK™ test is a membrane enzyme immunoassay that uses monoclonal and polyclonal antibodies to cell-surface antigens of the organisms. The polyclonal antibodies are coupled to horseradish peroxidase which are contained in the diluent. Specific mouse monoclonal antibodies against Giardia and Cryptosporidium antigens are bound on different location of the membrane. A fecal specimen is added to the diluent and transferred to the sample well and incubated. During the incubation, cyst and/or oocyst antigens in the sample bind the antibody-peroxidase conjugates. The antigen-antibody-conjugate complexes migrate through a filter pad to a membrane where they are captured by the immobilized Giardia and/or Cryptosporidium-specific antibodies in the test lines. ## M. Performance Characteristics (if/when applicable): 1. Analytical performance: a. Precision/Reproducibility: A total of 22 fecal specimens were pre-characterized by commercially available predicate devices. The samples included six Giardia-positive specimens (three mid-range positives), six Cryptosporidium-positive {4} specimens (three low positives), and four Giardia/Cryptosporidium-positive specimens (two of which were low Giardia-positives and two of which were low Cryptosporidium-positives), and six specimens negative for both parasites. All specimens were coded to prevent their identification during testing. Testing was performed at three sites. The samples were tested, twice a day over a 5-day period by multiple technicians at each site using two different kit lots. A positive and negative control was run with each panel of the masked samples. The results were consistent among the different locations, and exhibited a correlation of 100%. b. Linearity/assay reportable range: Not applicable c. Traceability, Stability, Expected values (controls, calibrators, or methods): Not applicable. d. Detection limit: The analytical sensitivity of the GIARDIA/CRYPTOSPORIDIUM QUIK CHEK test was determined by spiking various dilutions of purified Giardia cysts or Cryptosporidium oocysts into negative human stool. The cyst/oocyst preparations were quantified using the reference assay of IFA microscopy. Test results determined the limit of detection to be 6000 Giardia cysts per mL (133 cysts/test) and 6000 Cryptosporidium oocysts/mL (133 cysts/test). e. Analytical specificity: Cross- Reactivity Study No cross reactions were seen with the following organisms: Aeromonas hydrophila, Bacillus cereus, Bacillus subtilis, Bacteroides fragilis, Campylobacter coli, Campylobacter fetus, Campylobacter jejuni, Candida albicans, Clostridium bifermentans, Clostridium difficile (strain 630), Enterococcus faecalis, Escherichia coli, Escherichia coli 0157:H7, Escherichia coli ETEC (enterotoxic), Escherichia coli EPEC (enteropathogenic), Escherichia coli EIEC (enteroinvasive), Klebsiella pneumoniae, Salmonella typhimurium, Shigella dysenteriae, Shigella flexneri, Shigella sonnei, Staphylococcus aureus, Staphylococcus aureus (Cowan's), Staphylococcus epidermidis, Vibrio parahaemolyticus, Yersinia enterocolitica Human Adenovirus 1 and 3, Adenovirus, Type 2, 5, 40 and 41, Human Coxsackievirus B2, B3, and B4, Coxsackievirus B5, Human Coronavirus, Echovirus 11, 18, 33, Human Echovirus 9, Human paraechovirus 1 (Echovirus 22), Enterovirus 68, 69, Human Enterovirus 70, 71, Human rotavirus 5 {5} Additionally, the GIARDIA/CRYPTOSPORIDIUM QUIK CHEK™ test was tested using fecal specimens documented to be positive for other parasites by microscopy; *Ascaris lumbricoides* eggs, *Dientamoeba fragilis*, *Entamoeba hartmanni*, *Blastocystis hominis*, *Diphyllobothrium latum* eggs, *Entamoeba histolytica/E. dispar*, *Chilomastix mesnili*, *Endolimax nana*, Hookworm eggs, *Cyclospora cayetanensis*, *Entamoeba coli*, *Iodamoeba butschlii*, and *Trichuris trichiura* eggs. No cross-reactivity was seen with the following organisms for either the *Giardia*-portion or the *Cryptosporidium*-portion of the test. Cross-reactivity to Astrovirus and Caliciviruses has not been evaluated. The results obtained from the cross reactive study is acceptable. ## Interference Study The following substances had no effect on positive or negative test results analyzed at the concentrations indicated: Hog gastric mucin (3.5% w/v), Human blood (40% v/v), Barium sulfate (5% w/v), Imodium (5% v/v), Kaopectate (5% v/v), Pepto-Bismol (5% v/v), Steric/Palmitic Acid (40% w/v), Metronidazole (0.25% w/v), Vancomycin (0.25% w/v). The results obtained from the interference study is acceptable. ### f. Assay cut-off: Not Applicable ### 2. Comparison studies: #### a. Method comparison with predicate device: The performance of the GIARDIA/CRYPTOSPORIDIUM QUIK CHEK test was evaluated at three geographically diverse sites. Two sites compared the performance to commercially available ELISA (separate devices for *Giardia* and *Cryptosporidium*). A total of 849 specimens were evaluated and included 349 fresh, 322 frozen, 36 preserved (formalin) and 142 preserved (SAF). Two hundred and fifteen (215) specimens were positive for *Giardia* by ELISA. The GIARDIA/CRYPTOSPORIDIUM QUIK CHEK test exhibited a 99.1% agreement for *Giardia* positive specimens, a 99.7% agreement for *Giardia* negative specimens, with an overall agreement of 99.5%. One hundred thirty one (131) specimens were positive for *Cryptosporidium* by ELISA. The GIARDIA/CRYPTOSPORIDIUM QUIK CHEK test exhibited a 99.2% agreement for *Cryptosporidium* positive specimens, 99.6% agreement for *Cryptosporidium* negative specimens, and an overall 6 {6} agreement of 99.5%. The results obtained from the method comparison study are acceptable. b. Matrix comparison: Not Applicable 3. Clinical studies: a. Clinical Sensitivity: The performance of the GIARDIA/CRYPTOSPORIDIUM QUIK CHEK test was evaluated at three geographically diverse sites. Two sites compared performance to the reference microscopy/immunofluorescence (IFA). A total of 791 were evaluated and included 220 fresh, 140 frozen, 216 preserved (formalin) and 215 preserved (SAF). One hundred eighty three (183) specimens were positive for Giardia by IFA. The GIARDIA/CRYPTOSPORIDIUM QUIK CHEK test exhibited a sensitivity of 98.9%, a specificity of 100%, and an overall correlation of 99.7% with microscopy. One hundred forty (140) specimens were positive for Cryptosporidium by IFA. The GIARDIA/CRYPTOSPORIDIUM QUIK CHEK test exhibited a sensitivity of 100%, a specificity of 99.8%, and an overall correlation of 99.9% with Microscopy. The results obtained from the clinical study appear to be acceptable. Combined Results - Clinical Performance Comparing the Giardia Line of the GIARDIA/CRYPTOSPORIDIUM QUIK CHEK Test to Microscopy - IFA | N = 791 | Microscopy - IFA Giardia positive | Microscopy - IFA Giardia negative | | --- | --- | --- | | GIARDIA/CRYPTOSPORIDIUM QUIK CHEK Giardia Line Positive | 181 | 0 | | GIARDIA/CRYPTOSPORIDIUM QUIK CHEK Giardia Line Negative | 2 | 608 | | | | 95% Confidence Limits | | --- | --- | --- | | Sensitivity | 98.9% | 95.7 – 99.8% | | Specificity | 100% | 99.2 – 100% | | Correlation | 99.7% | 99.7 – 99.7% | {7} 8 # Combined Results - Clinical Performance Comparing the Cryptosporidium Line of the GIARDIA/CRYPTOSPORIDIUM QUIK CHEK Test to Microscopy – IFA | N = 791 | Microscopy - IFA Crypto. positive | Microscopy - IFA Crypto. negative | | --- | --- | --- | | GIARDIA/CRYPTOSPORIDIUM QUIK CHEK Cryptosporidium Line Positive | 140 | 1 | | GIARDIA/CRYPTOSPORIDIUM QUIK CHEK Cryptosporidium Line Negative | 0 | 650 | | | | 95% Confidence Limits | | --- | --- | --- | | Sensitivity | 100% | 96.7 – 100% | | Specificity | 99.8% | 99.0 – 100% | | Correlation | 99.9% | 100 – 100% | b. Clinical specificity: See section M.3.a c. Other clinical supportive data (when a. and b. are not applicable): Not Applicable 4. Clinical cut-off: Not Applicable 5. Expected values/Reference range: Normal healthy individuals should not be infected with Giardia or Cryptosporidium and should test negative in the GIARDIA/CRYPTOSPORIDIUM QUIK CHEK test. A positive test result in the GIARDIA/CRYPTOSPORIDIUM QUIK CHEK test indicates that the person is shedding detectable amounts of Giardia and/or Cryptosporidium antigen. The incidence of Giardia and Cryptosporidium infection varies significantly between populations and geographic regions. Children in daycare settings have exhibited higher rates of infection with Giardia than the normal population. In addition, homosexual men have shown higher rates of infection. In general, laboratory-confirmed incidence of cryptosporidiosis in developed countries ranges from 1 to 2% overall with a higher incidence in children. Prospective studies to evaluate the GIARDIA/CRYPTOSPORIDIUM QUIK CHEK test were conducted at an Asian hospital specializing in diarrheal diseases. The studies included 129 freshly collected fecal specimens. A positive test result in the {8} GIARDIA/CRYPTOSPORIDIUM QUIK CHEK test was obtained in 22.5% for Giardia and in 5.4% for Cryptosporidium antigen. N. Proposed Labeling: The labeling is sufficient and satisfies the requirements of 21 CFR Part 809.10. O. Conclusion: The submitted information in this premarket notification is complete and supports a substantial equivalence decision. 9