FaStep Early Pregnancy Rapid Test Strip is used for qualitative detection of Human Chorionic Gonadotropin (HCG) in human urine. This test is intended for use as an aid in early detection of pregnancy, in some cases as early as six (6) days before the day of the missed period, (i.e. as early as five (5) days before the day of the expected period). For over thecounter use.

FaStep Early Pregnancy Rapid Test Cassette is used for qualitative detection of Human Chorionic Gonadotropin (HCG) in human urine. This test is intended for use as an aid in early detection of pregnancy, in some cases as early as six (6) days before the day of the missed period, (i.e. as early as five (5) days before the day of the expected period). For over thecounter use.

FaStep Early Pregnancy Rapid Test Midstream is used for qualitative detection of Human Chorionic Gonadotropin (HCG) in human urine. This test is intended for use as an aid in early detection of pregnancy, in some cases as early as six (6) days before the day of the missed period, (i.e. as early as five (5) days before the day of the expected period). For overthe-counter use.

Device Description

FaStep Early Pregnancy Rapid Test will be sold in Strip, Cassette and Midstream format. The Strip format is a single test strip. The Cassette format consists of a single test strip assembled in a plastic housing. The Midstream format consists of a single test strip assembled in a plastic housing with an absorbent tip, and is designed to be tested in dip or midstream mode.

FaStep Early Pregnancy Rapid Test Strip, FaStep Early Pregnancy Rapid Test Cassette and FaStep Early Pregnancy Rapid Test Midstream each contains a pouch with the device and instructions, and in addition, cassette format is packaged with pipette dropper.

The devices utilize a combination of antibodies to detect hCG in urine as well as to serve as a run control. Each device contains mouse monoclonal anti-ß-hCG antibody colloidal gold conjugate pre-dried on the sample pad. Mouse monoclonal anti-a-hCG antibody (on the Test Line) and goat anti mouse IgG polyclonal antibody (on the Control Line) are coated and immobilized on a nitrocellulose membrane. The result is displayed to the user in the test window as two lines for a 'Pregnant' positive result and one line for a 'Not Pregnant' negative result.

AI/ML Overview

The provided document describes the performance and acceptance criteria for the FaStep Early Pregnancy Rapid Test Strip, Cassette, and Midstream devices.

Here's an analysis of the acceptance criteria and the study proving the device meets them:

1. Acceptance Criteria and Reported Device Performance

The acceptance criteria are implicitly derived from the performance claims made for the device and the comparison to the predicate device. The primary performance metrics revolve around sensitivity (detection limit for hCG) and accuracy (agreement with professional testing and predicate devices, and lack of interference/cross-reactivity).

Acceptance Criteria Category	Specific Acceptance Criteria (Implicit)	Reported Device Performance
Analytical Sensitivity	Detect 10 mIU/mL hCG with 100% positivity. No false positives at 0 or 5 mIU/mL hCG.	All three formats (Strip, Cassette, Midstream - both dip & in-stream methods) showed 0% positive at 0 and 5 mIU/mL hCG concentrations and 100% positive at 10 mIU/mL hCG and above.
Hook Effect	No hook effect up to 200,000 mIU/mL hCG.	No hook effect observed up to 200,000 mIU/mL hCG.
Interfering Substances	No interference from a specified list of common substances at given concentrations.	No interference observed at tested concentrations for all listed substances.
Cross-reactivity	No cross-reactivity with hLH, hFSH, and hTSH at specified concentrations.	No cross-reactivity observed with 500 mIU/mL hLH, 1000 mIU/mL hFSH, 1000 µIU/mL hTSH.
hCG ß-core fragment effect	Performance not affected by hCGBcf up to 500,000 pmol/L.	Performance not affected by hCGBcf concentrations up to 500,000 pmol/L.
Urine pH effect	Performance not affected by urine pH between 4 and 9.	Performance not affected by urine pH ranges between 4 and 9.
Urine Density effect	Performance not affected by urine density between 1.000 and 1.035.	Performance not affected by urine with a relative density range of 1.000 to 1.035.
Method Comparison (Professional Use)	100% conformity with the predicate device for positive and negative samples.	The conformity between FaStep Early Pregnancy Rapid Test (strip/cassette/midstream) and the predicate device was 100%.
Lay Person Use (Accuracy)	100% positive and 100% negative conformity with professional results in direct comparison. High percentage agreement with professional results for spiked samples (e.g., 100% at 5 and 10 mIU/mL, ≥97% at challenge concentrations like 7.5 and 8 mIU/mL).	First study: 100% positive and 100% negative conformity with professional results. Second study (spiked samples): 100% agreement at 5 mIU/mL and 10 mIU/mL. For 7.5 mIU/mL, agreement ranged from 97% to 98%. For 8.0 mIU/mL, agreement ranged from 98% to 99%.
Early Pregnancy Detection Rate	Detect early pregnancy consistent with the claimed "six (6) days before the day of the missed period" (EMP-5). Specifically, the clinical claim mentions "as early as five (5) days before the day of the expected period)" (EMP-5).	Detection rate in EMP-5 was 69.1% across all formats. Detection rate in EMP-4 was 89.7%. Detection rate in EMP-1 was 100%.
False-Positive Rate (Specificity)	No false positive results in non-pregnant females across various age groups.	No false positive results observed in 900 non-pregnant females (pre-menopausal, peri-menopausal, post-menopausal) across all three device formats and three lots. (0+/100- per age group per format/lot).

2. Sample Size and Data Provenance

Test Set Sample Sizes:
- Analytical Sensitivity (Precision/Reproducibility): For each format (Strip, Cassette, Midstream), 9 hCG concentrations tested. Each concentration had 10 replicates per day for 5 days, across 3 device lots, with 3 different operators. This sums to (9 concentrations * 10 replicates * 5 days * 3 lots * 3 operators = 1350 tests per format and per method for Midstream). More simply, the tables show 150 total results for each concentration level (50 per operator/lot). So, 9 concentrations * 150 results = 1350 tests per format.
- Hook Effect: Multiple hCG concentrations (6,250 to 200,000 mIU/mL). Specific number of replicates not stated, but results demonstrated.
- Interfering Substances: 3 hCG concentrations (0, 5, 10 mIU/mL) spiked with 25 substances. Number of replicates per substance not stated.
- Cross-reactivity: 3 hCG concentrations (0, 5, 10 mIU/mL) spiked with 3 cross-reactants. Number of replicates per cross-reactant not stated.
- hCG ß-core fragment effect: 4 hCG concentrations (0, 5, 10, 20,000 mIU/mL) spiked with 4 hCGBcf concentrations.
- Urine pH effects: 3 hCG concentrations (0, 5, 10 mIU/mL) tested at 6 pH values.
- Urine Density effects: 3 hCG concentrations (0, 5, 10 mIU/mL) tested at 8 density values.
- Method Comparison: 321 urine samples collected from women.
  - Strip format: 110 samples.
  - Cassette format: 105 samples.
  - Midstream format (dip): 106 samples.
  - Midstream format (in-stream): 106 samples.
- Lay Person Study (First Study - direct comparison): Total of 321 females.
  - Strip format: 110 subjects.
  - Cassette format: 105 subjects.
  - Midstream format (dip): 53 subjects.
  - Midstream format (in-stream): 53 subjects.
- Lay Person Study (Second Study - spiked samples): Total of 300 laypersons (100 per device format). Each tested 4 blind-labeled samples. So, 400 tests per format.
- Early Pregnancy Detection Study: 68 pregnant women provided 680 early pregnancy urine samples (from day -9 to 0 relative to expected period, presumably 10 samples per woman across these days). Each sample was tested with each format.
- Specificity Study (False-Positive Rate): 900 urine samples collected from non-pregnant females. 300 participants per age group (pre-menopausal, peri-menopausal, post-menopausal). For each age group, 100 tested strip, 100 tested cassette, 100 tested midstream (method not specified, but likely implies dip or in-stream was representative, or it was split). Three batches of each format were used.
Data Provenance:
- Country of Origin: Not explicitly stated, but clinical studies were conducted at "three clinical sites," which typically implies the country where the manufacturer is located or seeking approval (likely USA given FDA submission).
- Retrospective or Prospective:
  - Analytical studies (sensitivity, interference, etc.) are inherently prospective as they involve specific preparation and testing of samples.
  - Method comparison, lay person studies, early pregnancy detection study, and specificity study likely involved prospective collection of samples and testing as described (e.g., "Urine samples were collected from 321 women... at three clinical sites," "A lay user study was performed at three sites," "680 early pregnancy urine samples... were collected from 68 pregnant women," "900 urine samples were collected from non-pregnant females").

3. Number of Experts and Qualifications for Ground Truth

Ground Truth for Analytical Studies (Sensitivity, Interfering substances, etc.): The ground truth for hCG concentration was established by spiking negative female urine with hCG standard traceable to the 5th WHO (World Health Organization) standard. This indicates a highly standardized and globally recognized reference. The "operators" who performed the tests are not explicitly described as "experts" in establishing ground truth, but rather trained personnel conducting the tests.
Ground Truth for Method Comparison & Lay Person Studies (First Study): Ground truth was established by professional testing (laboratory professionals) and comparison to a predicate device. The document refers to "at least one professional at each site using the candidate device and by one professional at each site using the predicate device." Their specific qualifications are not detailed beyond "professional."
Ground Truth for Lay Person Study (Second Study - Spiked Samples): The samples were blind labeled by the person who prepared the samples (and did not take part in testing). The ground truth for the spiked samples was their known hCG concentration. These spiked urine samples were also tested by professionals to generate professional results for comparison with layperson results.
Ground Truth for Early Pregnancy Detection Study: The "positive results of B-ultrasound" (beta-ultrasound) served as the definitive ground truth for confirmed pregnancy at specific days relative to the expected menstrual period (EMP). This is a widely accepted clinical method for confirming pregnancy and gestational age.
Ground Truth for Specificity Study: "Non-pregnant females" were tested. The ground truth for "non-pregnant" would inherently be established by clinical assessment, possibly including absence of a positive ultrasound and absence of HCG in other clinical tests at the time of study enrollment.

4. Adjudication Method for the Test Set

No explicit "adjudication method" like 2+1 or 3+1 is mentioned for reconciling discordant results in the human studies.
For the analytical studies (e.g., sensitivity), reproducibility involves comparing results across multiple operators and lots.
For the method comparison and lay person studies, results are simply compared to the professional result or predicate device result. For the second lay person study, samples were "blind-labeled" with known concentrations, minimizing the need for adjudication of ground truth, but discrepancies between layperson and professional results are simply reported (e.g., "Percent Agreement").
For the early pregnancy detection study, results were correlated with B-ultrasound findings, which serves as a definitive clinical ground truth, reducing the need for consensus-based adjudication of the test results themselves.

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

No MRMC comparative effectiveness study was done. This device is a rapid diagnostic test (HCG test strip/cassette/midstream) for direct visual reading by a single user (professional or layperson), not an imaging device that would typically involve multiple readers interpreting complex images. Therefore, a study comparing human readers with and without AI assistance is not applicable.

6. Standalone (Algorithm Only) Performance

This is not an AI/algorithm-based device. It is a visually read immunoassay. Therefore, a standalone performance study of an algorithm is not applicable. The device's performance is inherently tied to the user's visual interpretation, even for "professional" use. However, the "Analytical performance" section could be considered the "standalone" performance in the sense of the device's inherent capability under controlled conditions, demonstrating sensitivity and specificity based on the chemical reaction.

7. Type of Ground Truth Used

The types of ground truth used varied by study:

Known hCG concentrations: For analytical sensitivity, hook effect, interfering substances, cross-reactivity, and effects of pH/density. These were generated in a lab setting by spiking negative urine samples with certified hCG standards.
Results from a legally marketed predicate device: For the method comparison study.
Professional testing/interpretation: For comparison in the lay person studies.
B-ultrasound (beta-ultrasound): For the early pregnancy detection study, serving as the definitive clinical confirmation of pregnancy.
Clinical status of "non-pregnant": For the specificity study, based on a cohort of non-pregnant females.

8. Sample Size for the Training Set

This document describes a 510(k) premarket notification for an in vitro diagnostic (IVD) test kit, not an AI/ML software device. As such, there is no "training set" in the context of machine learning model development. The data presented here are for validation/testing of the device's performance against its claims to establish substantial equivalence.

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

As explained in point 8, there is no "training set" for an AI/ML model for this type of IVD device. The various ground truths used for the validation studies are described in point 7.

Summary

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January 15, 2025

Image /page/0/Picture/1 description: The image shows the logo of the U.S. Food and Drug Administration (FDA). The logo consists of two parts: the Department of Health & Human Services logo on the left and the FDA logo on the right. The FDA logo is in blue and includes the letters "FDA" in a square, followed by the words "U.S. FOOD & DRUG ADMINISTRATION".

Assure Tech LLC % Jenny Xia Director LSI International Inc 504E Diamond Ave., Suite H Gaithersburg, Maryland 20877

Re: K243573

Trade/Device Name: FaStep Early Pregnancy Rapid Test Strip; FaStep Early Pregnancy Rapid Test Cassette; FaStep Early Pregnancy Rapid Test Midstream Regulation Number: 21 CFR 862.1155 Regulation Name: Human Chorionic Gonadotropin (HCG) Test System Regulatory Class: Class II Product Code: LCX Dated: November 18, 2024 Received: November 19, 2024

Dear Jenny Xia:

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.

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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).

Your device is also subject to, among other requirements, the Quality System (QS) 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.

All medical devices, including Class I and unclassified devices and combination product device constituent parts are required to be in compliance with the final Unique Device Identification System rule ("UDI Rule"). The UDI Rule requires, among other things, that a device bear a unique device identifier (UDI) on its label and package (21 CFR 801.20(a)) unless an exception or alternative applies (21 CFR 801.20(b)) and that the dates on the device label be formatted in accordance with 21 CFR 801.18. The UDI Rule (21 CFR 830.300(a) and 830.320(b)) also requires that certain information be submitted to the Global Unique Device Identification Database (GUDID) (21 CFR Part 830 Subpart E). For additional information on these requirements, please see the UDI System webpage at https://www.fda.gov/medical-device-advicecomprehensive-regulatory-assistance/unique-device-identification-system-udi-system.

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-regulatory

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assistance/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,

Joseph A. Digitally signed by
Joseph A. Joseph A. Kotarek -S Kotarek -S Date: 2025.01.15
10:19:37 -05'00'

Joseph Kotarek, Ph.D. Branch Chief 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) K243573

Device Name

FaStep Early Pregnancy Rapid Test Strip; FaStep Early Pregnancy Rapid Test Cassette; FaStep Early Pregnancy Rapid Test Midstream

Indications for Use (Describe)

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

K243573

1.	Date:	November 18, 2024
2.	Submitter:	Assure Tech. LLC1521 Concord Pike, Suite 201Wilmington, Delaware, 19803
3.	Contact person:	Jenny XiaLSI International Inc.504 East Diamond Ave., Suite HGaithersburg, MD 20877Telephone: 301-525-6856Email: jxa@lsi-consulting.org
4.	Device Name:	FaStep Early Pregnancy Rapid Test StripFaStep Early Pregnancy Rapid Test CassetteFaStep Early Pregnancy Rapid Test Midstream
	Classification:	Class II
	Product Code:	LCX
	CFR:	862.1155
5.	Predicate Devices:	Wondfo One Step HCG Urine Pregnancy Test MidstreamWondfo One Step HCG Urine Pregnancy Test Strip,Wondfo One Step HCG Urine Pregnancy Test Cassette(K150022)

6. Intended Use

FaStep Early Pregnancy Rapid Test Midstream is used for qualitative detection of

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Human Chorionic Gonadotropin (HCG) in human urine. This test is intended for use as an aid in early detection of pregnancy, in some cases as early as six (6) days before the day of the missed period, (i.e. as early as five (5) days before the day of the expected period). For over-the-counter use.

7. Device Description

Similarities
Item	Candidate device	Predicate device
Intended use	Early detection of pregnancy	Same
Specimen	Urine	Same
Assay technical	Immunochromatographicassay	Same
Sensitivity	10 mIU/mL	Same
Results	Qualitative	Same
Device format	Strip, Cassette, Midstream	Same
Differences
Item	Device	Predicate
Target user	For over-the-counter use	Prescription use and OTC use
Read time	3-10 minutes	5 minutes

8. Substantial Equivalence Information

Test Principle 9.

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FaStep Early Pregnancy Rapid Test Strip, FaStep Early Pregnancy Rapid Test Cassette and FaStep Early Pregnancy Rapid Test Midstream use lateral flow immunoassay for in vitro qualitative detection of Human Chorionic Gonadotropin (HCG) in human urine. If hCG is present in the sample, it will reach the Test Zone (T) of the membrane and form a colored line. When the test is performed properly, a colored line will always appear in the Control Zone (C). The test result is shown in the result window and read visually in 3-10 minutes of urine addition. Two distinct colored lines, one in the Test Zone (T) and another in the Control Zone (C) indicate a positive test result (pregnant). Absence of a colored line in the Test Zone (T) and only a colored line in the Control Zone (C) indicates a negative test result (not pregnant). Absence of a colored line in the Control Zone(C) even in the presence of a colored line in the Test Zone (T) indicates an invalid test result.

10. Performance Characteristics

A. Analytical performance

a. Precision/Reproducibility/Sensitivity

Negative female urine was spiked with hCG standard (Traceable to the 5th WHO) to hCG concentrations of 0, 5, 7.5, 8, 10, 15, 20, 25 and 50 mIU/mL. Each sample was tested in 10 replicates per day for 5 days for each device lot. Total of three device lots for each format were tested and each operator tested one lot separately. Tests were performed by three different operators for each sample concentration.

The midstream format was performed using both dip and in-stream methods.

The results are summarized in the table below:

hCGConcentration(mIU/mL)	Operator 1Lot 1		Operator 2Lot 2		Operator 3Lot 3		Total result		%Negative	%Positive
	-	+	-	+	-	+	-	+
0	50	0	50	0	50	0	150	0	100%	0%
5	50	0	50	0	50	0	150	0	100%	0%
7.5	31	19	29	21	30	20	90	60	60.0%	40.0%
8	24	26	25	25	24	26	73	77	48.7%	51.3%
10	0	50	0	50	0	50	0	150	0%	100%
15	0	50	0	50	0	50	0	150	0%	100%
20	0	50	0	50	0	50	0	150	0%	100%
25	0	50	0	50	0	50	0	150	0%	100%
50	0	50	0	50	0	50	0	150	0%	100%

Strip format

Cassette format

hCGConcentration(mIU/mL)	Operator 1Lot 1		Operator 2Lot 2		Operator 3Lot 3		Total result		% Negative	% Positive
	-	+	-	+	-	+	-	+
0	50	0	50	0	50	0	150	0	100%	0%
5	50	0	50	0	50	0	150	0	100%	0%
7.5	29	21	30	20	30	20	89	61	59.3%	40.7%
8	25	25	23	27	24	26	72	78	48.0%	52.0%
10	0	50	0	50	0	50	0	150	0%	100%

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15	50	50	50	150	0%	100%
20	50	50	50	150	0%	100%
25	50	50	50	150	0%	100%
50	50	50	50	150	0%	100%

Midstream format (in-stream method)

hCGConcentration(mIU/mL)	Operator 1Lot I		Operator 2Lot II		Operator 3Lot III		Total result		%Negative	%Positive
	-	+	-	+	-	+	-	+
	0	50	0	50	0	50	0	150
5	50	0	50	0	50	0	150	0	100%	0%
7.5	29	21	30	20	29	21	88	62	58.7%	41.3%
8	25	25	25	25	24	26	74	76	49.3%	50.7%
10	0	50	0	50	0	50	0	150	0%	100%
15	0	50	0	50	0	50	0	150	0%	100%
20	0	50	0	50	0	50	0	150	0%	100%
25	0	50	0	50	0	50	0	150	0%	100%
50	0	50	0	50	0	50	0	150	0%	100%

Midstream format (dip method)

hCGConcentration(mIU/mL)	Operator 1Lot I		Operator 2Lot II		Operator 3Lot III		Total result		% Negative	% Positive
	-	+	-	+	-	+	-	+
0	50	0	50	0	50	0	150	0	100%	0%
5	50	0	50	0	50	0	150	0	100%	0%
7.5	30	20	28	22	31	19	89	61	59.3%	40.7%
8	26	24	24	26	25	25	75	75	50.0%	50.0%
10	0	50	0	50	0	50	0	150	0%	100%
15	0	50	0	50	0	50	0	150	0%	100%
20	0	50	0	50	0	50	0	150	0%	100%
25	0	50	0	50	0	50	0	150	0%	100%
50	0	50	0	50	0	50	0	150	0%	100%

FaStep Early Pregnancy Rapid Test exhibited reproducibility of results.

Based on the above results, the sensitivity of FaStep Early Pregnancy Rapid Test is demonstrated to be 10 mIU/mL.

b. Linearity/assay reportable range:

Linearity is not applicable since this is a qualitative test. The test device was evaluated for high dose or hook effect.

Hook effect test:

Negative urine samples were spiked with varying hCG concentrations (6,250 mIU/mL, 12,500 mIU/mL, 25,000 mIU/mL, 50,000 mIU/mL, 100,000 mIU/mL, and 200,000 mIU/mL). The results demonstrated that no hook effect was observed at hCG concentration up to 200,000 mIU/mL.

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c. Traceability, Stability, Expected values (controls, calibrators, or methods): Traceability:

FaStep Early Pregnancy Rapid Test is calibrated against reference material traceable to WHO International Standard 5th edition, NIBSC code 07/364.

Stability:

The shelf-life of the FaStep Early Pregnancy Rapid Test at 2~30°C is 24 months based on real time stability data.

d. Analytical specificity

Interfering substance:

To evaluate potential interfering substances of the FaStep Early Pregnancy Rapid Test, urine samples containing 0 mIU/mL, 5 mIU/mL and 10 mIU/mL hCG were spiked with interfering substances to the concentrations listed below. No interference effect was observed at the tested concentrations.

Substance	Concentration
Acetaminophen	20 mg/dL
Acetylsalicylic Acid	20 mg/dl
Albumin	20 mg/dL
Amoxicillin	20 mg/dL
Ampicillin	20 mg/dL
Ascorbic acid	20 mg/dL
Aspirin	80 mg/dL
Atropine	20 mg/dL
Benzoylecgonine	10 mg/dL
Bilirubin	2 mg/dL
Caffeine	20 mg/dL
Cannabinol	10 mg/dL
Codeine	0.006 mg/dL
EDTA	80 mg/dL
Ephedrine	20 mg/dL
Ethanol	0.1%
Folic acid	0.03 mg/dL
Gentisic acid	20 mg/dL
Glucose	2000 mg/dL
Hemoglobin	20 mg/dL
Ibuprofen	40 mg/dL
Ketone	20 mg/dL
Methanol	1%
Phenothiazine	20 mg/dL
Phenylpropanolamine	20 mg/dL
Pregnanediol	1.5 mg/dL
Salicylic Acid	20 mg/dL

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Substance	Concentration
Tetracycline	20 mg/dL
Thiophene	20 mg/dL
Uric acid	23.5 mg/dL
Vitamin B1	80 mg/dL
ß-hydroxybutyrate	2000 mg/dL

Cross-reactivity :

To evaluate cross-reactivity, negative and positive urine samples (0 mIU/mL, 5 mIU/mL and 10 mIU/mL hCG) were spiked with potential cross reactants (500 mIU/mLhLH, 1000 mIU/mL hFSH, 1000 µIU/mL hTSH). No cross-reactivity was observed at tested concentration.

Effects of hCG ß-core fragment:

To evaluate the effect of the hCG ß-core fragment, negative urine samples (0 mIU/mL and 5 mIU/mL hCG) and positive urine samples (10 mIU/mL and 20,000 mIU/mL hCG) were spiked with hCG ß-core fragment (hCGBcf) at concentrations of 50.000 pmol/L, 125.000 pmol/L, 250.000 pmol/L and 500.000 pmol/L. The performance of FaStep Early Pregnancy Rapid Test was not affected by hCG ßcore fragment concentrations up to 500,000 pmol/L.

Effects of urine pH:

To evaluate the effect of urine pH on the results of FaStep Early Pregnancy Rapid Test, urine samples containing 0 mIU/mL, 5 mIU/mL and 10 mIU/mL hCG were tested at pH values of 4, 5, 6, 7, 8 and 9. The results indicated that urine pH ranges between 4 and 9 does not affect the performance of FaStep Early Pregnancy Rapid Test.

Effects of urine density:

To evaluate the effect of urine density on the results of FaStep Early Pregnancy Rapid Test, urine samples containing 0 mIU/mL, 5 mIU/mL and 10 mIU/mL hCG were tested at density values of 1.000, 1.005, 1.010, 1.015, 1.020, 1.025, 1.030 and 1.035. The results indicated that urine with a relative density rang of 1.000 to 1.035 does not affect the performance of FaStep Early Pregnancy Rapid Test.

B. Method comnarison study

Method comparison with predicate device:

The performance of the candidate device was compared to the predicate device. Urine samples were collected from 321 women aged 18 to 50 at three clinical sites. Approximately half of the subjects were pregnant in the early stage of less than 5 weeks. Samples were randomly collected at various time throughout the day.

All samples were tested by at least one professional at each site using the candidate device and by one professional at each site using the predicate device. 110 samples

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were tested for strip format, and 105 samples were tested for cassette format; for the midstream format, both dip testing and in-stream testing were evaluated for 106 samples. The samples were blinded and randomized before being tested by professionals. All results are summarized in the table below.

	Predicate device
Candidate device	Positive	Negative	Total
Strip format	Positive	61	0	61
	Negative	0	49	49
	Total	61	49	110

	Predicate device
Candidate device	Positive	Negative	Total
Cassette format	54	0	54
	0	51	51
Total	54	51	105

Candidate device		Predicate device
		Positive	Negative	Total
Midstream format(dip method)	Positive	57	0	57
	Negative	0	49	49
	Total	57	49	106

		Predicate device
Candidate device		Positive	Negative	Total
Midstream format(in-stream method)	Positive	57	0	57
	Negative	0	49	49
	Total	57	49	106

The conformity between FaStep Early Pregnancy Rapid Test (strip / cassette / midstream) and the predicate device is 100%.

C. Clinical studies

1. Clinical Sensitivity:
  Not applicable.
1. Clinical Specificity:
  Not applicable.
1. Other Clinical Supportive Data (When 1. and 2. Are Not Applicable):
3.1 Lay person study

First study:

A lay user study was performed at three sites with a total of 321 females with diverse educational and occupational backgrounds and ages ranging from 18 to 50

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years old. Each subject tested her own urine sample using the device according to the instructions for use and provided a sample for professional testing. 110 subjects tested the strip format, 105 subjects tested the cassette format, 53 subjects tested the midstream format using dip method, and 53 subjects tested the midstream format using in-stream method.

The results are summarized in the table below.

Strip format

	Professional
Strip format		Positive	Negative	Total
Layperson	Positive	61	0	61
	Negative	0	49	49
	Total	61	49	110

Cassette format

	Professional
Cassette format	Positive	Negative	Total
Layperson	Positive	54	0	54
	Negative	0	51	51
	Total	54	51	105

Midstream format

Midstream format(dip method)	Professional
	Positive	Negative	Total
Layperson	Positive	32	0	32
	Negative	0	21	21
	Total	32	21	53

Midstream format(in-stream method)	Professional
	Positive	Negative	Total
Layperson	Positive	25	0	25
	Negative	0	28	28
	Total	25	28	53

From the above tables, the lay person results showed 100% positive and 100% negative conformity with the professional results.

Second study:

Negative urine sample pools were spiked with 5 mIU/mL, 7.5 mIU/mL, 8 mIU/mL and 10 mIU/mL hCG. All aliquots were blind labeled by the person who prepared the samples and didn't take part in the sample testing. A total of 300 laypersons performed the testing, of which 100 using the strip format, 100 using the cassette format and 100 using the midstream format. Each layperson tested 4 blind-labeled

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samples (5 mIU/mL, 7.5 mIU/mL, 8 mIU/mL and 10 mIU/mL), and the spiked urine samples were also tested by professionals.

Results of spiked samples are summarized in the table below.

Strip format:

hCGConcentration(mIU/mL)	Layperson result			Professional result			PercentAgreement
	No. ofPositive	No. ofNegative	%positive	No. ofPositive	No. ofNegative	%positive
5	0	100	0%	0	100	0%	100%
7.5	39	61	39%	41	59	41%	98%
8.0	51	49	51%	53	47	53%	98%
10	100	0	100%	100	0	100%	100%

Cassette format:

hCGConcentration(mIU/mL)	Layperson result			Professional result			PercentAgreement
	No. ofPositive	No. ofNegative	%positive	No. ofPositive	No. ofNegative	%positive
5	0	100	0%	0	100	0%	100%
7.5	40	60	40%	42	58	42%	98%
8.0	51	49	51%	52	48	52%	99%
10	100	0	100%	100	0	100%	100%

Midstream format:

hCGConcentration(mIU/mL)	Layperson result			Professional result			PercentAgreement
	No. ofPositive	No. ofNegative	%positive	No. ofPositive	No. ofNegative	%positive
5	0	100	0%	0	100	0%	100%
7.5	39	61	39%	42	58	42%	97%
8.0	49	51	49%	51	49	51%	98%
10	100	0	100%	100	0	100%	100%

The results above show that FaStep Early Pregnancy Rapid Test can be used by the laypersons and get the correct results.

3.2 Early pregnancy detection study

A study was performed to evaluate the early pregnancy detection rate of FaStep Early Pregnancy Rapid Test. A total of 680 early pregnancy urine samples from day -9 to 0 relative to the day of the expected menstrual period were collected from 68 pregnant women. Each sample was tested with each format of the FaStep Early Pregnancy Rapid Test. For the midstream format, both dip and in-stream methods were evaluated.

The specific detection rates per day are summarized in the table below.

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Strip format:
Test date	EMP-9	EMP-8	EMP-7	EMP-6	EMP-5	EMP-4	EMP-3	EMP-2	EMP-1	EMP
# of positiveresults of B-ultrasound	68	68	68	68	68	68	68	68	68	68
# of positiveresults of HCG	0	5	10	28	47	61	66	67	68	68
Detection rate	0%	7.4%	14.7%	41.2%	69.1%	89.7%	97.1%	98.5%	100%	100%

Strin format:

Cassette format:

Test date	EMP-9	EMP-8	EMP-7	EMP-6	EMP-5	EMP-4	EMP-3	EMP-2	EMP-1	EMP
# of positiveresults of B-ultrasound	68	68	68	68	68	68	68	68	68	68
# of positiveresults of HCG	0	5	10	27	47	62	66	67	68	68
Detection rate	0%	7.4%	14.7%	39.7%	69.1%	91.2%	97.1%	98.5%	100%	100%

Midstream format (in-stream method):

Test date	EMP-9	EMP-8	EMP-7	EMP-6	EMP-5	EMP-4	EMP-3	EMP-2	EMP-1	EMP
# of positiveresults of B-ultrasound	68	68	68	68	68	68	68	68	68	68
# of positiveresults of HCG	0	5	10	27	47	61	66	67	68	68
Detection rate	0%	7.4%	14.7%	39.7%	69.1%	89.7%	97.1%	98.5%	100%	100%

Midstream format (dip method):

Test date	EMP-9	EMP-8	EMP-7	EMP-6	EMP-5	EMP-4	EMP-3	EMP-2	EMP-1	EMP
# of positiveresults of B-ultrasound	68	68	68	68	68	68	68	68	68	68
# of positiveresults of HCG	0	5	10	27	47	61	66	67	68	68
Detection rate	0%	7.4%	14.7%	39.7%	69.1%	89.7%	97.1%	98.5%	100%	100%

The detection rate of FaStep Early Pregnancy Rapid Test (strip/cassette/ midstream) in EMP-5 (i.e. five days before the expected menstrual period, 6 days before the missed menstrual period) was 69.1%, in EMP-4 was 89.7%, and in EMP-1 was 100%.

3.3 Specificity Study to Determine False-Positive Results Rate

900 urine samples were collected from non-pregnant females in pre-menopausal (18-40 years old), peri-menopausal (41-55 years old) and post-menopausal (>55 years old) age groups at three sites. 300 participants for each age group. In each age group, 100 participants test strip, 100 participants tested test cassette, 100 participants test midstream using dip method, or in-stream method. Three batches of each format were used in the study. No false positive results were

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observed for all age groups.

Strip format:

Group	Lot 1	Lot 2	Lot 3	Total result
Pre-menopausal	0+/34-	0+/33-	0+/33-	0+/100-
Peri-menopausal	0+/33-	0+/34-	0+/33-	0+/100-
Post-menopausal	0+/33-	0+/33-	0+/34-	0+/100-

Cassette format:

Group	Lot 1	Lot 2	Lot 3	Total result
Pre-menopausal	0+/34-	0+/33-	0+/33-	0+/100-
Peri-menopausal	0+/33-	0+/34-	0+/33-	0+/100-
Post-menopausal	0+/33-	0+/33-	0+/34-	0+/100-

Midstream format:

Group	Lot 1	Lot 2	Lot 3	Total result
Pre-menopausal	0+/34-	0+/33-	0+/33-	0+/100-
Peri-menopausal	0+/33-	0+/34-	0+/33-	0+/100-
Post-menopausal	0+/33-	0+/33-	0+/34-	0+/100-

11. Conclusion

Based on the test principle and performance characteristics of the device including precision, cut-off, interference, specificity, method comparison, lay-user study and early pregnancy detection study of the devices, it can be concluded that FaStep Early Pregnancy Rapid Test is substantially equivalent to the predicate device.

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.