The hCG One Step Pregnancy Test is an in vitro diagnostic test for the qualitative determination of human chorionic gonadotropin hormone in human urine samples. It is used as an aid in early detection of pregnancy, in some cases as early as five (5) days before the expected period, i.e., as early as six (6) days before the day of the missed period.

Device Description

The hCG One Step Pregnancy Test is a lateral flow sandwich immunochromatographic assay for the qualitative determination of human chorionic gonadotropin (hCG) in human urine samples.

AI/ML Overview

This report describes the acceptance criteria and the study that proves the device, the hCG One Step Pregnancy Test, meets those criteria.

Note: The provided document is a 510(k) summary, which focuses on demonstrating substantial equivalence to a predicate device, rather than providing a detailed clinical study report typically associated with AI/ML-based devices. Therefore, some of the requested information, particularly regarding ground truth establishment by experts, adjudication, and MRMC studies, is not explicitly detailed as this device is a qualitative lateral flow immunoassay, not an AI-assisted diagnostic.

1. Acceptance Criteria and Reported Device Performance

The acceptance criteria for this device are demonstrated through analytical performance studies (Limit of Detection, Precision/Reproducibility, Analytical Specificity) and a comparison study to a legally marketed predicate device.

Acceptance Criteria Category	Specific Criteria/Study Goal	Reported Device Performance
Analytical Performance
Limit of Detection (LOD)	Determine the lowest concentration of hCG that yields ≥ 95% positive results. The target sensitivity is 10 mIU/mL.	10 mIU/mL for urine obtained by both simulated stream method and dip method. This was confirmed by testing serial dilutions of hCG standard substance with 3 lots of devices by 3 technicians over multiple days. At 10 mIU/mL and above, 100% positive rate was obtained.
Precision/Reproducibility	Show consistent results across multiple operators, lots, and days, especially at and around the LOD.	When testing hCG at 10 mIU/mL and above, a 100% positive rate was obtained for all operators with all device lots, at both the simulated stream method and the dip method. The device produced consistent results.
Analytical Specificity
Interfering Substances	No interference from common prescription/OTC drugs, elevated chemical analytes (caffeine, ascorbic acid), or elevated biological analytes (glucose, protein, albumin, bilirubin, hemoglobin) at relevant levels.	No interference observed from the potential interfering substances at the levels evaluated when testing negative urine samples and samples with 10 mIU/mL hCG.
Cross-Reactivity	No cross-reactivity with substances structurally similar to hCG.	Compounds tested (not explicitly named but implied to be similar in structure) at approximate 0 and 10 mIU/mL hCG concentrations were found to have no cross-reactivity.
Comparative Performance
Method Comparison	Demonstrate comparable performance to a legally marketed predicate device (First Response Early Result Pregnancy Test) using clinical urine samples.	The test performance of the hCG One Step Pregnancy Test showed 100% concordance when compared to the predicate device. This was evaluated by professional users at 2 sites using 158 clinical urine samples.

2. Sample Sizes and Data Provenance

Test Set Sample Size:
- Limit of Detection: The panel was created by spiking 30 negative urine samples with hCG. The number of individual test runs was significantly higher, as 3 lots of the device were tested by 3 lab technicians on different days, with randomized, blind-coded panel members. The specific total count of test runs is not provided but is a multiple of 30.
- Precision/Reproducibility: Twenty replicates of each spiked urine sample (concentrations 0, 5, 7.5, 8.75, 10, 12.5, and 25 mIU/mL) were tested in one day for repeatability. For intermediate precision, 450 replicates per urine sample concentration were tested (10 replicates per operator x 3 operators x 3 lots x 5 nonconsecutive days).
- Analytical Specificity (Interfering Substances): Five devices were used for testing each interfering substance.
- Method Comparison: A total of 158 urine samples from 158 subjects.
Data Provenance:
- The document does not explicitly state the country of origin for the clinical urine samples.
- The studies appear to be prospective as per standard FDA premarket notification requirements for device testing. The "urine samples were collected at various times of day in a random fashion" and "masked with a code and randomized by individuals who were not conducting the test," suggesting a structured study design.

3. Number of Experts and Qualifications for Ground Truth

This information is not explicitly detailed in the document.
For this type of qualitative immunoassay (pregnancy test), the "ground truth" for the test set is established by the presence or absence of hCG in the urine samples at specified concentrations, verified through the spiking process using traceable hCG standards (5th WHO international Standard) and comparison to a predicate device.
The interpretation of the device results during testing was performed by "lab technicians" for analytical studies and "professional users" for the comparison study. Their specific qualifications (e.g., years of experience, specific medical background) are not provided, but these roles imply trained personnel capable of accurately performing and interpreting the test. No expert radiologists or similar specialists are required for this device type.

4. Adjudication Method for the Test Set

Not applicable/Not explicitly stated in the context of expert adjudication.
For the analytical studies (LOD, Precision), the ground truth is based on the known concentration of hCG in the spiked samples. The device's performance is then measured against this known concentration.
For the method comparison study, the comparison is made against the results of the legally marketed predicate device. Discrepancies (if any, though 100% concordance was reported) would likely prompt re-evaluation of the sample or test, rather than a formal expert adjudication process as seen in imaging studies. The samples were "masked with a code and randomized by individuals who were not conducting the test," which helps prevent bias in interpretation.

5. Multi-Reader Multi-Case (MRMC) Comparative Effectiveness Study

No, an MRMC comparative effectiveness study was not performed. This type of study is typically associated with AI/ML-based diagnostic imaging devices where human readers interpret medical images with and without AI assistance.
The hCG One Step Pregnancy Test is a qualitative immunoassay, where the result is visually interpreted (presence/absence of a line). The "performance" is about whether the line appears correctly for a given hCG concentration, not about optimizing human interpretation of complex data.
While multiple operators (lab technicians, professional users) were involved in testing to demonstrate reproducibility and comparability, this is distinct from an MRMC study assessing AI-assisted human reader improvement.

6. Standalone Performance (Algorithm Only)

Yes, in essence, the device's performance is standalone.
The hCG One Step Pregnancy Test is a non-AI, non-software-based diagnostic device. Its performance (e.g., sensitivity, specificity, limit of detection) is inherent to its physical and chemical properties and is evaluated directly through the analytical and comparison studies.
There is no "algorithm only" component separate from the physical test strip and its reagents. The results are based on the immunochromatographic reaction itself.

7. Type of Ground Truth Used

Analytical Ground Truth (for LOD, Precision, Analytical Specificity): Based on known concentrations of hCG in spiked urine samples, traceable to the 5th WHO international Standard. This is a highly controlled and quantitative ground truth.
Comparative Ground Truth (for Method Comparison): The results from a legally marketed predicate device (First Response Early Result Pregnancy Test) using the same clinical urine samples. This establishes equivalence to an accepted standard.

8. Sample Size for the Training Set

Not applicable/Not provided. This device is not an AI/ML device that requires a distinct "training set." Its chemical and biological components are designed and manufactured, not "trained" on data. The studies described are for validation and verification of the finished product's performance.

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

Not applicable. As stated above, there is no training set for this type of medical device.

Summary

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June 30, 2022

Healstone Biotech Inc. % Guang Gao Principal Consultant Axteria Biomed Consulting 8040 Cobble Creek Circle Potomac, Maryland 20854

Re: K212418

Trade/Device Name: hCG One Step Pregnancy Test Regulation Number: 21 CFR 862.1155 Regulation Name: Human Chorionic Gonadotropin (HCG) Test System Regulatory Class: Class II Product Code: LCX Dated: May 5, 2022 Received: May 5, 2022

Dear Guang Gao:

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. 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 located 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.

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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 803) for devices or postmarketing safety reporting (21 CFR 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safety-reportingcombination-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 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

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 https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.

For comprehensive regulatory information about medical devices and radiation-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.

Marianela Perez-Torres, Ph.D. Deputy Director Division of Chemistry and Toxicology Devices OHT7: Office of In Vitro Diagnostics Office of Product Evaluation and Quality Center for Devices and Radiological Health

Enclosure

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Indications for Use

510(k) Number (if known) K212418

Device Name hCG One Step Pregnancy Test

Indications for Use (Describe)

Type of Use (Select one or both, as applicable)
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Prescription Use (Part 21 CFR 801 Subpart D)

|X | Over-The-Counter Use (21 CFR 801 Subpart C)

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K212418

510(K) SUMMARY

Submitter's Identification: Healstone Biotech Inc. Unit 650, 655 West Kent Ave. N., Vancouver, BC, V6P 6T7, Canada www.orientgene.com

Date: June 29, 2022

Contact Person:

Guang Gao, Principal Consultant Axteria Biomed Consulting 8040 Cobble Creek Circle Potomac, MD 20854 (301) 814-4985 ggw101413@icloud.com

Proprietary Name of the Device: hCG One Step Pregnancy Test

Common Name: hCG One Step Pregnancy Test

Classification Name: 21 CFR § 862.1155 Human chorionic gonadotropin (HCG) test system

Product Code: LCX

1.0 Device Description

The hCG One Step Pregnancy Test is a lateral flow sandwich immunochromatographic assay for the qualitative determination of human chorionic gonadotropin (hCG) in human urine samples.

2.0 Intended Use

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3.0 Test Principle

The hCG One Step Pregnancy Test is a lateral flow sandwich immunochromatographic assay that utilizes a combination of monoclonal antibodies for qualitative detection of hCG in human urine samples. The test uses two lines to show the results. When the urine sample is added into the absorbent tip, the sample migrates via capillary action along the membrane. If present in the sample, hCG will react with the specific monoclonal hCG antibody and form a colored line at the test line region of the membrane, indicating a positive result (pregnant). Absence of a colored line at the test line region suggests a negative result (not pregnant). To serve as a procedural control, a colored line will always appear in the control line region. If the control line does not appear (no color appears at the control line region 3 minutes after application of the sample), the test is invalid and the specimen should be retested with a new device.

Similarities and Differences
	Proposed device	Predicate device
Name	hCG One Step Pregnancy Test	First Response Early Result Pregnancy Test (K123436)
Device Type	In vitro diagnostic	Same
Classification	Class II	Same
Regulatory Number	21 CFR §862.1155 Human Chorionic Gonadotropin	Same
Product Usage	OTC	Same
Principle of Operation	Lateral flow sandwich immunochromatographic assay	Same
Specimen Collection Method	Mid-stream or dip mode	Same
Intended Use	A chromatographic immunoassay for the qualitative detection of human chorionic gonadotropin (hCG) in urine. The device is intended for use as an aid in early detection of pregnancy, in some cases as early as five (5) days before the expected period, i.e., as early as six (6) days before the day of the missed period.	Same

4.0 Comparison of the Predicate Device

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Type of Specimen	Human urine	Same
Sensitivity	10 mIU/mL	Same
Time to Result	3 minutes	Same
Result	Qualitative	Same
Shelf Life	24 months shelf life when stored ina dry place at 2-30°C.	Same
hCG IsoformsDetected	Intact hCG	Intact hCG,hyperglycosylated hCG, hCGß-subunit, hCG ß-corefragment

Test Performance 5.0

Analytical Performance 5.1

Limit of Detection 5.1.1
The Limit of Detection (LOD) was determined by testing serial dilutions of the hCG standard substance that is commercially available and traceable to the 5th WHO international Standard. Three internal sites used 3 lots of the proposed device with a simulated stream method and dip method. The panel was created by spiking 30 negative urine samples (from males and nonpregnant females) with the hCG standard substance at eight different hCG concentrations (below, at and above the initial assigned LOD). These panel members were randomized, blind coded and tested with the 3 lots of the proposed device by three different lab technicians on different days. Each operator performed the test with one lot of the proposed device and interpreted the results independently.

The results showed that the LOD (the lowest concentration that yields ≥ 95% positive results) is 10 mIU/mL for urine obtained by both the simulated stream method and the dip method.

Precision/Reproducibility 5.1.2

Panels were prepared from 100% pooled human urine containing no analyte. The hCG human urine samples were spiked with hCG positive specimen that is traceable to the 5th WHO international standard to obtain the hCG at concentrations 0, 5, 7.5, 8.75, 10, 12.5, and 25 mIU/mL.

Repeatability testing was conducted with twenty replicates of each urine sample in one day with one lot of the hCG One Step Pregnancy Test. For intermediate precision testing, each panel member was tested by three operators with 10 replicates and 3 lots of device for 5 nonconsecutive days. A total of 450 replicates of each urine sample was tested over multiple days by multiple operators. When testing hCG at 10 mIU/mL and above, a 100% positive rate was

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obtained for all operators with all device lots, at both the simulated stream method and the dip method.

The results show that the proposed device can produce consistent results when detecting hCG in urine samples.

Analytical Specificity 5.1.3

5.1.3.1 Potential Interfering Substances Study

The interference substance study was performed with urine containing prescription/OTC drugs, elevated levels of chemical analytes (e.g., caffeine, ascorbic acid), and elevated levels of biological analytes (e.g., glucose, protein, albumin, bilirubin, hemoglobin).

For the potential interfering substances study, negative female urine samples containing 0 and 10 mIU/mL of hCG were individually spiked with the potential interfering substances. Five devices were used for the testing. Based on the five replicates results, no interference was observed from the potential interfering substances at the level evaluated.

5.1.3.2 Cross Reactivity Study

The hCG One Step Pregnancy Test was evaluated for cross-reactivity with other substances that are similar in structure to hCG. Human urine specimens with hCG concentrations of approximately 0 and 10 mIU/mL were supplemented with potentially cross-reacting compounds. The compounds were tested and found to have no cross-reactivity.

5.2 Comparison Study

The performance the hCG One Step Pregnancy Test was evaluated by professional users at 2 sites with 2 lots of the proposed device. A total of 158 urine samples were collected from 158 subjects with ages ranging between 18 to 45 who were in need of determining their pregnancy status. The urine samples were collected at various times of day in a random fashion. The urine specimens were tested with the proposed device and with the predicate device (First Response Early Result Pregnancy Test) by independent operators. The urine samples were masked with a code and randomized by individuals who were not conducting the test. The test results of the hCG One Step Pregnancy Test and the First Response Early Result Pregnancy Test were compared to evaluate the clinical test performance.

The test performance of the hCG One Step Pregnancy Test is with 100% concordance when compared to the predicate.

6.0 Conclusion

Based on the test principle and acceptable performance characteristics including interfering substances, cross reactivity, method comparison, clinical specificity, and lay-user studies of the devices, it is concluded that the hCG One Step Pregnancy Test is substantially equivalent to the predicate.

Regulation Number and Section

§ 862.1155 Human chorionic gonadotropin (HCG) test system.

(a)
Human chorionic gonadotropin (HCG) test system intended for the early detection of pregnancy —(1)Identification. A human chorionic gonadotropin (HCG) test system is a device intended for the early detection of pregnancy is intended to measure HCG, a placental hormone, in plasma or urine.(2)
Classification. Class II.(b)
Human chorionic gonadotropin (HCG) test system intended for any uses other than early detection of pregnancy —(1)Identification. A human chorionic goadotropin (HCG) test system is a device intended for any uses other than early detection of pregnancy (such as an aid in the diagnosis, prognosis, and management of treatment of persons with certain tumors or carcinomas) is intended to measure HCG, a placental hormone, in plasma or urine.(2)
Classification. Class III.(3)
Date PMA or notice of completion of a PDP is required. As of the enactment date of the amendments, May 28, 1976, an approval under section 515 of the act is required before the device described in paragraph (b)(1) may be commercially distributed. See § 862.3.