(88 days)
Not Found
No
The device description and performance studies detail a standard enzyme immunoassay (ELISA) method for detecting H. pylori antigens. There is no mention of AI or ML in the intended use, device description, or performance evaluation sections. The analysis involves spectrophotometric readings and comparison to established cut-off values, which are not indicative of AI/ML processing.
No.
The device is an in vitro diagnostic qualitative enzyme immunoassay intended to aid in the diagnosis and post-therapy diagnosis of H. pylori infection by detecting antigens. It does not provide any treatment or therapy.
Yes
Explanation: The "Intended Use / Indications for Use" section explicitly states, "The Hp Detect™ Stool Antigen ELISA is an in vitro diagnostic qualitative enzyme immunoassay..." and "The Hp Detect™ Stool Antigen ELISA is intended to aid in the initial diagnosis and post-therapy diagnosis of H. pylori infection."
No
The device is an in vitro diagnostic kit containing physical components such as a microplate, antibodies, buffers, and controls, which are used in a laboratory setting to perform an enzyme immunoassay. It is not solely software.
Yes, this device is an IVD (In Vitro Diagnostic).
The "Intended Use / Indications for Use" section explicitly states: "The Hp Detect™ Stool Antigen ELISA is an in vitro diagnostic qualitative enzyme immunoassay for the detection of Helicobacter pylori (H. pylori) antigens in human stool or feces."
This statement directly identifies the device as an in vitro diagnostic product.
N/A
Intended Use / Indications for Use
The Hp Detect Stool Antigen ELISA is an in vitro diagnostic qualitative enzyme immunoassay for the detection of Helicobacter pylori (H. pylori) antigens in human stool or feces. The Hp Detect Stool Antigen ELISA is intended to aid in the initial diagnosis and post-therapy diagnosis of H. pylori infection. Additionally, the test may be used to assess H. pylori infection status after treatment. Retesting at a minimum of 4 weeks after the completion of treatment may be done to assess H. pylori status. Test results should always be taken into consideration by the physician in conjunction with patient's clinical information (history and symptoms). For Prescription Use Only.
Product codes (comma separated list FDA assigned to the subject device)
LYR
Device Description
The Hp Detect Stool Antigen ELISA is an enzyme immunoassay which detects the H. pylori antigen in human fecal samples. The Hp Detect Stool Antigen ELISA comes in a kit that contains materials to assay a total of 92 samples. The device consists of a 96-well clear flat bottom polystyrene high bind microplate coated with affinity purified rabbit anti-human H. pylori polyclonal antibody. The device is provided with detection antibody which is a purified mouse monoclonal antibody specific for H. pylori antigen and has been conjugated to horseradish peroxidase (HRP). The device kit is also provided with sample diluent buffer, wash buffer, substrate solution, stop solution along with negative and positive controls. Negative control is a phosphate buffered protein solution and positive control is composed of purified H. pylori antigen (ATCC strain 43504) from cell lysate.
Polyclonal anti-H. pylori captures antibodies that are immobilized on microwells. Patient samples prepared in sample diluent are added to the microwells and incubated for one hour at 37 ± 2℃. If the H. pylori antigen is present in the sample, it will bind to the immobilized antibody on the plate. Following this incubation, the plate is washed thoroughly. A peroxidase conjugated anti-H. pylori monoclonal antibody is then added to the microwells and incubated for 30 minutes at 37 ± 2℃. If H. pylori antigen is bound to the microwells in the first step, the detection antibody would now bind in this step to form a sandwich complex. Following this incubation, a thorough wash step is performed to remove non-specific and non-binding materials. Substrate is then added and incubated for 10 minutes at 37±2℃ to generate a color in the presence of the enzyme complex. Stop solution is then added to end the reaction. The results are read spectrophotometrically at the following wavelengths:
-
- Single Wavelength Measurement at 450 nm
-
- Dual Wavelength Measurement 450/620 nm or 450/630 nm
Mentions image processing
Not Found
Mentions AI, DNN, or ML
Not Found
Input Imaging Modality
Not Found
Anatomical Site
Stool or feces (human)
Indicated Patient Age Range
Not Found
Intended User / Care Setting
Prescription Use Only.
Description of the training set, sample size, data source, and annotation protocol
Not Found
Description of the test set, sample size, data source, and annotation protocol
A panel of 227 specimens, of which 83 were positive and 144 were negative by the predicate device were evaluated with the Hp Detect Stool Antigen ELISA for assay cut-off determination.
For clinical study - frozen specimen: A total of 433 frozen and de-identified fecal samples were tested in which 355 specimens were collected from Italy and 78 specimens collected from three geographically different regions of the USA (west, southwest, and southeast). The clinical specimens were collected from the patients presenting with dyspepsia, and would be undergoing a routine endoscopy and biopsy, not on antibiotics, bismuth, or proton-pump inhibitors, and those excluded if they had been treated for H. pylori within the past 6 months. Specimens were distributed into three testing sites located within the USA which include two external sites and one internal (Biomerica) site.
For clinical study - fresh stool specimen: A total of 142 fresh, de-identified, fecal specimens from patients with physician's medical evaluation for symptoms of H. pylori infection were collected through multiple biospecimen vendor and clinical laboratories. The specimens were tested in the collection centers with the comparator device and specimen aliquots were shipped to Biomerica (internal site).
For post-therapy diagnosis: 14 paired (pre-and post- therapy) frozen retrospective specimens from Italy were used. All subjects initially tested positive for H. pylori by composite reference method (CRM) consisting of histology and Rapid Urease Test and all completed eradication therapy. Post-therapy fecal samples were collected at a minimum of 4 weeks post completion of eradication therapy.
Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)
Precision / Reproducibility:
Reproducibility study: A four (4) panel of specimens were prepared at high negative (0.42 × LoD), low positive (1.60 × LoD), low positive (2.43 × LoD) and moderate positive (3.93 × LoD) by spiking H. pylori antigen (ATCC strain 43504) into a negative pooled fecal matrix. The reproducibility panel was blinded, and each run included positive and negative controls. Testing was performed at three sites, two independent laboratories and one in-house at Biomerica, Inc. The samples were tested in triplicate twice per day over a 5-day period by two technicians at each site using one kit lot (2 Technicians × 3 sites × 3 replicates × 2 runs × 5 days = 180 results/panel member).
Results (Combined, 3 sites): High Negative (0.42 LoD): 7/180 Positive/Tested (3.9% Detection); Low Positive (1.60 LoD): 180/180 Positive/Tested (100% Detection); Low Positive (2.43 LoD): 180/180 Positive/Tested (100% Detection); Moderate Positive (3.93 LoD): 180/180 Positive/Tested (100% Detection). Both dual (450/620 or 450/630 nm) and single (450 nm) wavelength produced equivalent results.
Within-Lab precision/repeatability: A four (4) panel of specimens as prepared for the reproducibility study was used. Two operators performed the test with two validation lots per run over the period of 12 non-consecutive days. Two runs per day with a minimum of 3 hours separation between runs. Three replicates of each sample per run were tested along with positive and negative controls.
Results (Combined, Two kit lots): High Negative (0.42 LoD): 13/288 Positive/Tested (5% Detection); Low Positive (1.60 LoD): 278/288 Positive/Tested (97% Detection); Low Positive (2.43 LoD): 288/288 Positive/Tested (100% Detection); Moderate Positive (3.93 LoD): 288/288 Positive/Tested (100% Detection). Both dual (450/620 or 450/630 nm) and single (450 nm) wavelength produced equivalent results.
Analytical Specificity (Cross-Reactivity and Microbial Interference): The test was evaluated for cross-reactivity and microbial interference with known bacteria, fungi and viruses. A contrived positive matrix was prepared by spiking H. pylori purified antigen (ATCC strain 43504) at 2.2 × LoD (5.61 ng/mL) into the negative fecal matrix. Bacteria and fungi were spiked at concentrations of 10^6-10^7 CFU/mL and viruses at a range from 10^4-10^5 TCID50 units per mL. No cross-reactivity or microbial interference was observed.
Interfering Substances: The test was evaluated for interference with potential interfering substances (e.g., Barium Sulfate, Human Hemoglobin, Metronidazole, Mucin, Palmitic Acid, Polyethylene Glycol 3350, Steric Acid, Vancomycin Hydrochloride, Whole Blood, OTC Medications like Imodium AD, Kaopectate, Mylanta, Pepto Bismol, Prilosec, Simethicone, Tagamet, TUMS). No interference was observed.
Clinical Study - Frozen Specimen: Method comparison study with an FDA cleared device. Total of 433 frozen and de-identified fecal samples (355 from Italy, 78 from USA).
Dual wavelength results (450/620 or 450/630 nm): Positive Percent Agreement (PPA) 99.11% (95% CI: 95.12 - 99.84 %), Negative Percent Agreement (NPA) 98.13 % (95% CI: 95.98 - 99.14 %).
Single wavelength results (450 nm): PPA 99.11% (95% CI: 95.12 – 99.84 %), NPA 95.95 % (95% CI: 93.20 – 97.62 %).
Clinical Study - Fresh Stool Specimen: Method comparison study with an FDA cleared device. Total of 142 fresh, de-identified, fecal specimens. Measurement using both dual wavelength (450/620 or 450/630 nm) and single wavelength (450 nm) settings yielded the same results.
PPA 100.00% (95% CI: 83.89 - 100%), NPA 98.36% (95% CI: 94.22 - 99.55%).
Post -Therapy Diagnosis: Performance evaluated using 14 paired (pre-and post- therapy) frozen retrospective specimens from Italy, compared to composite reference method (CRM).
Sensitivity 100% (95% CI: 72.25 - 100 %), Specificity 100 % (95% CI: 51.02 - 100 %).
Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)
Clinical Study - Frozen Specimen (Dual Wavelength):
Positive Percent Agreement (PPA): 99.11%
Negative Percent Agreement (NPA): 98.13%
Clinical Study - Frozen Specimen (Single Wavelength):
Positive Percent Agreement (PPA): 99.11%
Negative Percent Agreement (NPA): 95.95%
Clinical Study - Fresh Stool Specimen (Dual and Single Wavelength):
Positive Percent Agreement (PPA): 100.00%
Negative Percent Agreement (NPA): 98.36%
Post -Therapy Diagnosis:
Sensitivity: 100%
Specificity: 100%
Predicate Device(s): If the device was cleared using the 510(k) pathway, identify the Predicate Device(s) K/DEN number used to claim substantial equivalence and list them here in a comma separated list exactly as they appear in the text. List the primary predicate first in the list.
Reference Device(s): Identify the Reference Device(s) K/DEN number and list them here in a comma separated list exactly as they appear in the text.
Not Found
Predetermined Change Control Plan (PCCP) - All Relevant Information for the subject device only (e.g. presence / absence, what scope was granted / cleared under the PCCP, any restrictions, etc).
Not Found
§ 866.3110
Campylobacter fetus serological reagents.(a)
Identification. Campylobacter fetus serological reagents are devices that consist of antisera conjugated with a fluorescent dye used to identifyCampylobacter fetus from clinical specimens or cultured isolates derived from clinical specimens. The identification aids in the diagnosis of diseases caused by this bacterium and provides epidemiological information on these diseases.Campylobacter fetus is a frequent cause of abortion in sheep and cattle and is sometimes responsible for endocarditis (inflammation of certain membranes of the heart) and enteritis (inflammation of the intestines) in humans.(b)
Classification. Class I (general controls).
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Image /page/0/Picture/0 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). On the left is the Department of Health & Human Services logo. To the right of that is the FDA logo, which is a blue square with the letters "FDA" in white. To the right of the square is the text "U.S. FOOD & DRUG ADMINISTRATION" in blue.
December 15, 2023
Biomerica, Inc. Bonnie Qiu Senior Director, Quality Assurance and Regulatory Affairs 17571 Von Karman Avenue Irvine, California 92614
Re: K232892
Trade/Device Name: Hp Detect Stool Antigen ELISA Regulation Number: 21 CFR 866.3110 Regulation Name: Campylobacter Fetus Serological Reagents Regulatory Class: Class I. reserved Product Code: LYR Dated: September 15, 2023 Received: September 18, 2023
Dear Bonnie Oiu:
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 (the 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. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database available at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. 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.
Additional information about changes that may require a new premarket notification are provided in the FDA guidance documents entitled "Deciding When to Submit a 510(k) for a Change to an Existing Device" (https://www.fda.gov/media/99812/download) and "Deciding When to Submit a 510(k) for a Software Change to an Existing Device" (https://www.fda.gov/media/99785/download).
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Your device is also subject to, among other requirements, the Quality System (OS) regulation (21 CFR Part 820), which includes, but is not limited to, 21 CFR 820.30, Design controls; 21 CFR 820.90, Nonconforming product; and 21 CFR 820.100, Corrective and preventive action. Please note that regardless of whether a change requires premarket review, the QS regulation requires device manufacturers to review and approve changes to device design and production (21 CFR 820.30 and 21 CFR 820.70) and document changes and approvals in the device master record (21 CFR 820.181).
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 Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR Part 803) for devices or postmarketing safety reporting (21 CFR Part 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safetyreporting-combination-products); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR Part 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR Parts 1000-1050.
Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.
For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).
Sincerely,
Natasha Griffin -S
0.B.O. Ribhi Shawar, Ph.D. (ABMM) Branch Chief General Bacteriology and Antimicrobial Susceptibility Branch Division of Microbiology Devices OHT7: Office of In Vitro Diagnostics Office of Product Evaluation and Quality Center for Devices and Radiological Health
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Indications for Use
510(k) Number (if known) K232892
Device Name Hp Detect™ Stool Antigen ELISA
Indications for Use (Describe)
The Hp Detect™ Stool Antigen ELISA is an in vitro diagnostic qualitative enzyme immunoassay for the detection of Helicobacter pylori (H. pylori) antigens in human stool or feces. The Hp Detect™ Stool Antigen ELISA is intended to aid in the initial diagnosis and post-therapy diagnosis of H. pylori infection. Additionally, the test may be used to assess H. pylori infection status after treatment. Retesting at a minimum of 4 weeks after the completion of treatment may be done to assess H. pylori status. Test results should always be taken into consideration by the physician in conjunction with patient's clinical information (history and symptoms). For Prescription Use Only.
| Type of Use (Select one or both, as applicable) | |
|---|---|
| Prescription Use (Part 21 CFR 801 Subpart D) | Over-The-Counter Use (21 CFR 801 Subpart C) |
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510(k) Summary
A. SUBMITTER/510(K) HOLDER
Biomerica, Inc 17571 Von Karman Ave Irvine, CA 92614
B. DEVICE NAME
| Proprietary Name: | Hp Detect Stool Antigen ELISA |
|---|---|
| Common/Usual Name: | Helicobacter pylori stool antigen ELISA |
| Classification Name: | Helicobacter pylori |
REGULATORY INFORMATION ﺰ
| Regulation Section: | 21 CFR 866.3110 Campylobacter fetus serological reagents |
|---|---|
| Classification: | Class I |
| Product Code: | LYR |
| Panel: | 83-Microbiology |
D. PREDICATE DEVICE PREMIER Platinum HpSA PLUS (K182559)
E. DEVICE DESCRIPTION
The Hp Detect Stool Antigen ELISA is an enzyme immunoassay which detects the H. pylori antigen in human fecal samples. The Hp Detect Stool Antigen ELISA comes in a kit that contains materials to assay a total of 92 samples. The device consists of a 96-well clear flat bottom polystyrene high bind microplate coated with affinity purified rabbit anti-human H. pvlori polyclonal antibody. The device is provided with detection antibody which is a purified mouse monoclonal antibody specific for H. pylori antigen and has been conjugated to horseradish peroxidase (HRP). The device kit is also provided with sample diluent buffer, wash buffer, substrate solution, stop solution along with negative and positive controls. Negative control is a phosphate buffered protein solution and positive control is composed of purified H. pylori antigen (ATCC strain 43504) from cell lysate.
Polyclonal anti-H. pylori captures antibodies that are immobilized on microwells. Patient samples prepared in sample diluent are added to the microwells and incubated for one hour at 37 ± 2℃. If the H. pvlori antigen is present in the sample, it will bind to the immobilized antibody on the plate. Following this incubation, the plate is washed thoroughly. A peroxidase conjugated anti-H. pylori monoclonal antibody is then added to the microwells and incubated for 30 minutes at 37 ± 2℃. If H. pylori antigen is bound to the microwells in the first step, the detection antibody would now bind in this step to form a sandwich complex. Following this incubation, a thorough wash step is performed to remove non-specific and non-binding materials. Substrate is then added and incubated for 10 minutes at 37±2℃ to generate a color in the presence of the enzyme complex. Stop solution is then added to end the reaction. The results are read spectrophotometrically at the following wavelengths:
4
-
- Single Wavelength Measurement at 450 nm
-
- Dual Wavelength Measurement 450/620 nm or 450/630 nm
F. INDICATION FOR USE/INTENDED USE
The Hp Detect Stool Antigen ELISA is an in vitro diagnostic qualitative enzyme immunoassay for the detection of Helicobacter pylori (H. pylori) antigens in human stool or feces. The Hp Detect Stool Antigen ELISA is intended to aid in the initial diagnosis and posttherapy diagnosis of H. pylori infection. Additionally, the test may be used to assess H. pylori infection status after treatment. Retesting at a minimum of 4 weeks after the completion of treatment may be done to assess H. pylori status. Test results should always be taken into consideration by the physician in conjunction with patient's clinical information (history and symptoms). For Prescription Use Only.
ﻥ SUMMARY OF TECHNOLOGICAL CHARACTERISTICS COMPARED TO THE PREDICATE DEVICE
A summary comparison of technological characteristics, including design and materials is provided in the following Table 1.
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| Item | Device: K232892 | Predicate : K182559 |
|---|---|---|
| Device | ||
| Trade Name | Hp Detect Stool Antigen ELISA | PREMIER Platinum HpSA PLUS |
| General Device Characteristic Similarities | ||
| Product | ||
| Code | LYR | Same |
| Intended | ||
| Use | The Hp Detect Stool Antigen | |
| ELISA is an in vitro diagnostic | ||
| qualitative enzyme immunoassay | ||
| for the detection of Helicobacter | ||
| pylori ( H. pylori ) antigens in | ||
| human stool or feces. The Hp | ||
| Detect Stool Antigen ELISA is | ||
| intended to aid in the initial | ||
| diagnosis and post therapy | ||
| diagnosis of H. pylori infection. | ||
| Additionally, the test may be used | ||
| to assess H. pylori infection status | ||
| after treatment. Retesting at a | ||
| minimum of 4 weeks after the | ||
| completion of treatment may be | ||
| done to assess H. pylori status. Test | ||
| results should always be taken into | ||
| consideration by the physician in | ||
| conjunction with patient's clinical | ||
| information (history and | ||
| symptoms). For Prescription Use | ||
| Only. | The PREMIER Platinum HpSA | |
| PLUS enzyme immunoassay | ||
| (EIA) is an in vitro qualitative | ||
| assay for the detection of | ||
| Helicobacter pylori antigens in | ||
| human stool. Test results are | ||
| intended to aid in the diagnosis | ||
| of H. pylori infection and to | ||
| monitor response during and | ||
| post-therapy in patients. | ||
| Accepted medical practice | ||
| recommends that testing by any | ||
| current method, to confirm | ||
| eradication, be done at least four | ||
| weeks following completion of | ||
| therapy. | ||
| Measured | ||
| Analyte | Detection of H. pylori antigen | Same |
| Type of Test | Qualitative | Same |
| Specimen | ||
| Type | Human fecal specimens | Same |
| Controls | Positive and negative control | |
| included in the kit | Positive and negative control | |
| included in the kit | ||
| Target | ||
| Population | Persons suspected of having H. | |
| pylori infection | Same | |
| Storage | Refrigerated (2°C – 8°C) | Same |
| Item | Device: K232892 | Predicate : K182559 |
| General Device Characteristic Differences | ||
| Specimen | ||
| Storage | Specimens may be held up to 120 | |
| hours at 2°C – 8°C or should be | ||
| frozen at ≤ -12°C or ≤ -60°C | Specimens may be held up to 72 | |
| hours at 2°C – 8°C or at | ||
| -20°C to -25°C prior to testing | ||
| Time to | ||
| Result | Approximately 1 hour and 40 | |
| minutes | Approximately 1 hour and 10 | |
| minutes | ||
| Antibody | ||
| Format | Polyclonal/Monoclonal | Monoclonal mixture/Monoclonal |
| mixture | ||
| Incubation | ||
| Temp | 37°C ± 2°C | 19-27°C |
| Reading | ||
| Method | Spectrophotometric | Visual, Spectrophotometric |
Table 1. Comparison with Predicate Device
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H. PERFORMANCE CHARACTERISTICS
1. Analytical Performance
a. Precision / Reproducibility
To demonstrate the reproducibility of the Hp Detect Stool Antigen ELISA, a four (4) panel of specimens were prepared at high negative (0.42 × LoD), low positive (1.60 × LoD), low positive (2.43 × LoD) and moderate positive (3.93 × LoD) by spiking H. pylori antigen (ATCC strain 43504) into a negative pooled fecal matrix. The reproducibility panel was blinded, and each run included positive and negative controls. Testing was performed at three sites, two independent laboratories and one in-house at Biomerica, Inc. The samples were tested in triplicate twice per day over a 5-day period by two technicians at each site using one kit lot (2 Technicians × 3 sites × 3 replicates × 2 runs × 5 days = 180 results/panel member). The results for the reproducibility study were measured using the dual wavelength (450/620 or 450/630 nm) and are summarized in Table 2 below. Testing at the dual and single wavelength (450 nm) produced equivalent results.
| Sample ID | H. pylori Antigen | Combined (3 sites) | |
|---|---|---|---|
| Positive/Tested | % Detection | ||
| High Negative | $0.42 \times \text{LoD}$ | 7/180 | 3.9% |
| Low Positive | $1.60 \times \text{LoD}$ | 180/180 | 100% |
| Low Positive | $2.43 \times \text{LoD}$ | 180/180 | 100% |
| Moderate Positive | $3.93 \times \text{LoD}$ | 180/180 | 100% |
Table 2. Reproducibility of the Hp Detect Stool Antigen ELISA measured in dual wavelength
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The Within-Lab precision/repeatability of the Hp Detect Stool Antigen ELISA was determined onsite at Biomerica using a four (4) panel of specimens as prepared for the reproducibility study as described above. Two operators performed the test with two validation lots per run over the period of 12 non-consecutive days. Two runs per day with a minimum of 3 hours separation between runs. Three replicates of each sample per run were tested along with positive and negative controls. The results for the Within-Lab precision/repeatability study measured in dual wavelength (450/620 or 450/630 nm) are summarized in Table 3. Testing at the dual and single wavelength (450 nm) produced equivalent results.
| Sample ID | H. pylori Antigen | Combined (Two kit lots) | |
|---|---|---|---|
| Positive/ Tested | % Detection | ||
| High Negative | $0.42 \times LoD$ | 13/288 | 5% |
| Low Positive | $1.60 \times LoD$ | 278/288 | 97% |
| Low Positive | $2.43 \times LoD$ | 288/288 | 100% |
| Moderate Positive | $3.93 \times LoD$ | 288/288 | 100% |
Table 3. Within-Lab precision/repeatability of the Hp Detect Stool Antigen ELISA measured in dual wavelength.
The results for reproducibility and precision/repeatability studies were acceptable.
b. Linearity / Assay Reportable Range
Not Applicable
c. Traceability, Stability, Expected Values (Controls, Calibrators, or Methods)
Traceability:
Calibrator traceability is not applicable (No calibrator is included in kit).
Sample Stability:
Sample Stability - Contrived Specimens
The sample stability for contrived specimens was evaluated for both refrigerated and frozen storage conditions including freeze and thaw. Samples were prepared by spiking H. pylori antigen (ATCC strain 43504) into a negative pooled fecal matrix. The sample panel was made up of 10 members of H. pylori antigen concentrations ranging from 0.47 to 5.42 × LoD tested in triplicates. After establishing the baseline measurement (time zero), each of the 10 panel members were aliquoted and stored at three storage conditions: 2℃ to 8℃. H. pylori Strain | Concentration | Positive/Tested
(% Detection) |
|-------------------------|--------------------------|----------------------------------|
| ATCC 43526 | $3.70 \times 103$ CFU/mL | 3/3 (100%) |
| ATCC 43579 | $3.48 \times 103$ CFU/mL | 3/3 (100%) |
| ATCC 49396 | $3.42 \times 103$ CFU/mL | 3/3 (100%) |
| ATCC 700392 | $3.85 \times 103$ CFU/mL | 3/3 (100%) |
| ATCC 700824 | $3.96 \times 103$ CFU/mL | 3/3 (100%) |
| ATCC BAA-945 | $3.78 \times 103$ CFU/mL | 3/3 (100%) |
| ATCC 49503 | 6.33 ng/mL Antigen | 3/3 (100%) |
Table 9. List of H. pylori strains tested for inclusivity study.
The Inclusivity results were acceptable.
f. Assay Cut-off
Assay cut-off for Hp Detect Stool Antigen ELISA was determined for both spectrophotometric measurement of Dual Wavelength (450/620 nm or 450/630 nm) and Single Wavelength (450 nm). A panel of 227 specimens, of which 83 were positive and 144 were negative by the predicate device were evaluated with the Hp Detect Stool Antigen ELISA. Receiver operating characteristic (ROC) curve was constructed and the cut-off values for spectrophotometric single and dual wavelength reading methods were selected to achieve optimum diagnostic sensitivity and specificity:
Spectrophotometric Dual Wavelength (450/620 nm or 450/630 nm) Negative: 0.140
A positive result indicates that H. pylori antigens were detected. A negative result indicates that no H. pylori antigens were detected, or that the antigen levels are below what can be detected by the assay.
g. Prozone / Hook Effect
To ensure that a high concentration of H. pylori antigen does not interfere with a positive reaction in the Hp Detect Stool Antigen ELISA, a high positive sample was prepared by spiking H. pylori antigen (ATCC strain 43504) into negative fecal matrix. The sample was serially diluted at concentrations ranging from 20,000 ng/mL to 0.25 ng/mL. Each of the serial dilutions were tested in 8 replicates measuring by both dual (450/620 or 450/630 nm) and single wavelength (450 nm) settings. Positive and negative controls were included in each run. The reading value of the plate reader reached to maximum level at 800 ng/ml of H. pylori antigen and remained at maximum level up to 20,000 ng/ml. No high-dose hook effect was observed for the assay at antigen concentrations of up to 20,000 ng/mL.
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2. Clinical Study
Clinical Study - Frozen Specimen
The performance of the Hp Detect Stool Antigen ELISA for frozen specimens was evaluated by using method comparison study with an FDA cleared device. A total of 433 frozen and deidentified fecal samples were tested in which 355 specimens were collected from Italy and 78 specimens collected from three geographically different regions of the USA (west, southwest, and southeast). The clinical specimens were collected from the patients presenting with dyspepsia, and would be undergoing a routine endoscopy and biopsy, not on antibiotics, bismuth, or proton-pump inhibitors, and those excluded if they had been treated for H. pylori withing the past 6 months. Specimens were distributed into three testing sites located within the USA which include two external sites and one internal (Biomerica) site. Results were evaluated by reading absorbance at dual wavelength (450/620 or 450/630 nm) and single wavelength (450 nm). The performance and statistics are shown in Table 10 for dual wavelength and Table 11 for single wavelength.
Table 10. Frozen Specimen - Clinical performance and statistics measured in dual wavelength.
| Hp Detect Stool Antigen ELISA | Comparator: FDA Cleared Device | ||
|---|---|---|---|
| Positive | Negative | Total | |
| Positive | 111 | 6a | 117 |
| Negative | 1b | 315 | 316 |
| Total | 112 | 321 | 433 |
| Statistics | Performance | 95% Confidence Interval |
|---|---|---|
| Positive Percent Agreement (PPA) | 99.11% | 95.12 - 99.84 % |
| Negative Percent Agreement (NPA) | 98.13 % | 95.98 - 99.14 % |
The discrepant results were further analyzed by chart review and determined to have a RUT or history result and observed as follow:
(a) Four of the six discrepant false positives were either comparator positive by RUT or histology.
(b) The discrepant false negative was negative by RUT or histology.
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| Hp Detect Stool Antigen ELISA | Comparator: FDA Cleared Device | ||
|---|---|---|---|
| Positive | Negative | Total | |
| Positive | 111 | 13a | 124 |
| Negative | 1b | 308 | 309 |
| Total | 112 | 321 | 433 |
| Statistics | Performance | 95% Confidence Interval | |
| Statistics | Performance | 95% Confidence Interval | |
| Positive Percent Agreement (PPA) | 99.11% | 95.12 – 99.84 % | |
| Negative Percent Agreement (NPA) | 95.95 % | 93.20 – 97.62 % |
Table 11. Clinical performance and statistics measured in single wavelength.
The discrepant results were further analyzed by chart review and determined to have a RUT or history result and observed as follow:
- (a) Four of the 13 discrepant false positive were positive by RUT or histology.
- (b) The discrepant false negative was negative by RUT or histology.
The performance results for frozen specimen clinical study are acceptable.
Clinical Studv - Fresh Stool Specimen
The performance of the Hp Detect Stool Antigen ELISA for fresh specimens (never frozen) was evaluated by using method comparison study with an FDA cleared device. A total of 142 fresh, de-identified, fecal specimens from patients with physician's medical evaluation for symptoms of H. pylori infection were collected through multiple biospecimen vendor and clinical laboratories. The specimens were tested in the collection centers with the comparator device and specimen aliquots were shipped to Biomerica (internal site) and tested using Hp Detect Stool Antigen ELISA using both dual wavelength (450/620 or 450/630 nm) and single wavelength (450 nm) settings.
| Table 12. Fresh Specimens - Clinical performance and statistics measured in dual and single | ||||
|---|---|---|---|---|
| wavelength with same results. | ||||
| Hp Detect Stool Antigen ELISA | Comparator: FDA Cleared Device | |||
| Positive | Negative | Total | ||
| Positive | 20 | 2 | 22 | |
| Negative | 0 | 120 | 120 | |
| Total | 20 | 122 | 142 |
| Statistics | Performance | 95% Confidence Interval |
|---|---|---|
| Positive Percent Agreement (PPA) | 100.00% | 83.89 - 100% |
| Negative Percent Agreement (NPA) | 98.36% | 94.22 - 99.55% |
The Hp Detect Stool Antigen ELISA demonstrated the clinical performance of 100% positive percent agreement and 98.4% negative percent agreement for fresh stool specimens using both dual and single wavelength configurations. The performance results for fresh specimen clinical study are acceptable.
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Post -Therapy Diagnosis
The performance of the Hp Detect Stool Antigen ELISA for post-therapy diagnosis was evaluated by using 14 paired (pre-and post- therapy) frozen retrospective specimens from Italy. All subjects initially tested positive for H. vvlori by composite reference method (CRM) consisting of histology and Rapid Urease Test and all completed eradication therapy. Posttherapy fecal samples were collected at a minimum of 4 weeks post completion of eradication therapy. All 14 pre-therapy samples were previously shown to be positive and matched with the CRM diagnosis. The result for post-therapy was shown in Table 13.
| Hp Detect Stool Antigen ELISA | Post-Therapy CRM | ||
|---|---|---|---|
| Positive | Negative | Total | |
| Positive | 10 | 0 | 10 |
| Negative | 0 | 4 | 4 |
| Total | 10 | 4 | 14 |
Table 13. Post-Therapy Performance with composite reference method
| Statistics | Performance | 95% Confidence Interval |
|---|---|---|
| Sensitivity | 100% | 72.25 - 100 % |
| Specificity | 100 % | 51.02 - 100 % |
The results for post-therapy performance show that the Hp Detect Stool Antigen ELISA exhibited a 100% sensitivity and 100% specificity when compared to the composite reference method. The performance results for post-therapy study are acceptable and support the claim for the test to be used in assessing H. pylori status after treatment.
Clinical Cut-Off
Not Applicable.
Expected Values – Reference Range
The Hp Detect Stool Antigen ELISA assay detects the presence of H. pylori antigen in human feces. The clinical study included in this submission had results from 433 frozen specimens collected from three independent sites in the US, each located in a different geographical region of the country - west, southwest, and southeast, and outside the US - Italy. The observed prevalence of H. pvlori in this study was 27.0% (117/433). H. pvlori detection and prevalence stratified by specimen origin are shown in Table 14 and categorized by gender is shown in Table 15.
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| Specimen Origin | Hp Detect Stool Antigen ELISA Result | | Total | Prevalence
(% Positive) |
|-----------------|--------------------------------------|----------|-------|----------------------------|
| | Negative | Positive | | |
| US - Southeast | 26 | 8 | 34 | 23.5% |
| US - Southwest | 13 | 3 | 16 | 18.8% |
| US - West | 19 | 9 | 28 | 32.1% |
| US (All Sites) | 58 | 20 | 78 | 25.6% |
| Italy | 258 | 97 | 355 | 27.3% |
| Grand Total | 316 | 117 | 433 | 27.0% |
Table 14: H. pylori detection and prevalence by specimen origin
Table 15: H. pylori detection and prevalence by gender
| Gender | Hp Detect Stool Antigen ELISA Result | | Total | Prevalence
(% Positive) |
|---------|--------------------------------------|----------|-------|----------------------------|
| | Negative | Positive | | |
| Females | 205 | 76 | 281 | 27.0% |
| Males | 111 | 41 | 152 | 26.9% |
| Total | 316 | 117 | 433 | 27.0% |
I. PROPOSED LABELING
The labeling supports the finding of substantial equivalence for this device.
J. CONCLUSIONS
The submitted information in this premarket notification is complete and supports substantial equivalence decision.