Wondfo One Step HCG Urine Pregnancy Test Strip is a rapid chromatographic immunoassay for the qualitative detection of human chorionic gonadotropin (bCG) in urine, 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. Important note regarding negative results: Some pregnant women will not be able to detect hCG in their urine 5 days before the expected period. If you test negative before your missed period, but think you may still be pregnant, you should test again a few days after your missed period. Important note regarding positive results: Because this test detects low levels of hCG, it is possible that this test may give positive results even if you are not pregnant. If you test positive, but think you may not be pregnant, you should check with your doctor. All results should be confirmed by your healthcare provider, especially when making decisions about future medical care. This product is intended for both prescription use and over-the-counter use.

Wondfo One Step HCG Urine Pregnancy Test Cassette is a rapid chromatographic immunoassay for the qualitative detection of human chorionic gonadotropin (hCG) in urine, 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. Important note regarding negative results: Some pregnant women will not be able to detect hCG in their urine 5 days before the expected period. If you test negative before your missed period, but think you may still be pregnant, you should test again a few days after your missed period. Important note regarding positive results: Because this test detects low levels of hCG, it is possible that this test may give positive results even if you are not pregnant. If you test positive, but think you may not be pregnant, you should check with your doctor. All results should be confirmed by your healthcare provider, especially when making decisions about future medical care. This product is intended for both prescription use and over-the-counter use.

Wondfo One Step HCG Urine Pregnancy Test Midstream is a rapid chromatographic immunoassay for the qualitative detection of human chorionic gonadotropin (hCG) in urine, 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. Important note regarding negative results: Some pregnant women will not be able to detect hCG in their urine 5 days before the expected period. If you test negative before your missed period, but think you may still be pregnant, you should test again a few days after your missed period. Important note regarding positive results: Because this test detects low levels of hCG, it is possible that this test may give positive results even if you are not pregnant. If you test positive, but think you may not be pregnant, you should check with your doctor. All results should be confirmed by your healthcare provider, especially when making decisions about future medical care. This product is intended for over-the-counter use.

Device Description

Each of the devices (strip, cassette, and midstream) contains a pouch with the test and instructions. The cassette and midstream nitrocellulose test strips are contained in plastic housing. The cassette test also contains a dropper. The strips of each device contain mouse monoclonal anti-β-hCG antibody colloidal gold conjugate pre-dried on the pad, mouse monoclonal anti-u-hCG antibody (on the Test Line) and goat anti mouse IgG polyclonal antibody (on the Control Line).

AI/ML Overview

Here's a breakdown of the acceptance criteria and study details for the Wondfo One Step HCG Urine Pregnancy Test, as extracted from the provided document:

1. Table of Acceptance Criteria and Reported Device Performance

The document does not explicitly state "acceptance criteria" in a tabulated format with pass/fail marks. However, it presents the performance characteristics measured and the results obtained for the device. Based on the information provided, we can infer the performance criteria and the device's reported performance:

Acceptance Criteria (Inferred)	Reported Device Performance
Analytical Sensitivity / Cut-Off Value: Ability to detect specific hCG concentrations (critical at 8 mIU/mL and 10 mIU/mL).	Cut-off value: 8 mIU/mL (established) Analytical sensitivity: 10 mIU/mL (established) Detection Rates: - 7.5 mIU/mL: ~28-30% positive across formats and lots. - 8 mIU/mL: ~50-51.7% positive across formats and lots. - 9 mIU/mL: ~88.3-90% positive across formats and lots. - 10 mIU/mL and above: 100% positive across formats and lots.
Stability: Maintain performance over time.	Stable at 4-30°C for 24 months (based on accelerated stability at 50°C and real-time stability at 4°C and 30°C).
Specificity and Cross-Reactivity: No interference from high hCG concentrations (no hook effect), different hCG fragments (B-core), or other glycoprotein hormones (LH, FSH, TSH).	High Dose Effect: No hook effect observed at hCG concentrations ranging from 6,250 to 200,000 mIU/mL. Effects of hCG ß-core fragment: No interference observed for urine samples with 0 mIU/mL hCG and 10 mIU/mL hCG (except false negative at 500,000 pmol/L of ß-core fragment hCG, which is a very high concentration). Effects of glycoprotein LH, FSH and TSH: No interference observed at LH concentrations up to 500 IU/mL, FSH concentrations up to 1000 mIU/mL, and TSH concentrations up to 1000 µIU/mL.
Interfering Substances: No interference from various common substances.	No interference observed for all 20 tested compounds (Acetaminophen, Acetylsalicylic acid, Ascorbic acid, Atropine, Caffeine, Gentisic acid, Glucose, Hemoglobin, Tetracycline, Ampicillin, Albumin, B-hydroxybutyrate, Ephedrine, Phenylpropanolamine, Phenothiazine, EDTA, Salicyclic Acid, Benzoylecgonine, Cannabinol, Codeine, Ethanol, Bilirubin, Atropine, Pregnanediol, Thiophene, Ketone) at stated concentrations, for both 0, 10, and 100 mIU/mL hCG samples.
Effect of Urine Specific Gravity and Urine pH: Consistent performance across varying urine properties.	No interference from pH ranging from 4 to 9. No interference from specific gravity ranging from 1.000 to 1.035.
Precision: Consistent results across multiple replicates, operators, and lots.	Consistent results across 10 replicates for 5 consecutive days by three different operators at 3 POC sites, using 3 different lots for each format, for hCG concentrations ranging from 0 to 50 mIU/mL. E.g., for 10 mIU/mL, all 50 replicates were positive across all 3 lots for all formats. For 0, 2.5, 5 mIU/mL, all 50 replicates were negative across all 3 lots for all formats. The detection rates for 7.5 mIU/mL and 8 mIU/mL show expected variability around the cut-off.
Method Comparison with Predicate Device: High agreement with a legally marketed predicate device.	100% agreement for both positive and negative samples when compared to the FIRST RESPONSE™ Early Result Pregnancy Test (K123436) for all three device formats (Strip, Cassette, Midstream).
Lay Person Study: Ease of use and accuracy when used by intended lay users.	Overall Agreement with Professional: - Strip: 100% agreement (49+/51- vs 49+/51-). - Cassette: 99% agreement (48+/51- vs 48+/52-), with 1 false positive by lay person. - Midstream: 100% agreement (49+/51- vs 49+/51-). Correctness for Spiked Samples (5mIU/ml, 10mIU/ml): - Strip: 99% for 5mIU/ml (1+/99-), 100% for 10mIU/ml (100+/0-). (Note: The table on page 13 for strips and 5 mIU/mL seems to have a typo, showing 1 positive for 5mIU/mL. The overall result for lay person vs professional for strips is 100% agreement, which contradicts the 1 positive result for 5mIU/mL in the next table. Assuming the professional comparison is the primary indicator of overall accuracy vs ground truth). - Cassette: 100% for 5mIU/ml (0+/100-), 100% for 10mIU/ml (100+/0-). - Midstream: 100% for 5mIU/ml (0+/100-), 99% for 10mIU/ml (99+/1-). Consistency for Spiked Samples (7.5mIU/mL, 8mIU/mL): - Strip: 96% and 97%. - Cassette: 96% and 96%. - Midstream: 96% and 97%. Ease of Use: All lay users indicated instructions were easily followed. Flesch-Kincaid reading Grade Level of 7 for package inserts.
False Positive Rate: Low rate of positive results in non-pregnant individuals.	Strip: 0.3% false positive results (1 false positive out of 300 non-pregnant females, from a peri-menopausal woman confirmed non-pregnant via ultrasound). Cassette: 0.3% false positive results (1 false positive out of 300 non-pregnant females, from a peri-menopausal woman confirmed non-pregnant via ultrasound). Midstream: 0% false positive results (0 out of 300 non-pregnant females).
Early Pregnancy Detection: Ability to detect pregnancy early relative to the Expected Menstrual Period (EMP).	Detects: - 68% positive hCG five days before EMP. - 100% positive hCG on the day of EMP. - Performance consistent across device formats.
Substantial Equivalence: Overall performance comparable to the predicate device.	Concluded to be substantially equivalent to the predicate device (Church & Dwight Co., Inc. FIRST RESPONSE™ Early Result Pregnancy Test).

2. Sample Size Used for the Test Set and Data Provenance

Analytical Performance (Sensitivity/Cut-Off, Interference, Precision):
- Sensitivity/Cut-Off: 20 replicates per hCG concentration level per lot (3 lots), tested by 12 operators. This means 60 tests per hCG level per format (e.g., for Strip format: 60 tests for 0mIU/ml, 60 for 5mIU/ml, etc.). The study used Urine standards containing intact hCG calibrated against WHO 4th IS for hCG. The provenance is not explicitly stated but is implied to be laboratory-prepared standards.
- Specificity & Cross-reactivity: Samples (negative urine and 10 mIU/mL hCG) spiked with specified interfering substances. Each spiked sample was tested by 3 different lots and 3 different operators. The exact number of replicates per sample/condition is not explicitly stated, but for each interfering substance, a series of tests were done to ensure no interference.
- Precision: 10 replicates per day for 5 consecutive days using 3 different lots for each format. This means 50 tests per hCG concentration per lot per format (e.g., for Strip format, 50 tests for 0mIU/ml, 50 for 2.5mIU/ml, up to 50 for 50mIU/ml, for each of the 3 lots). Tests were performed by 9 POC operators (3 for each format). The ground truth was negative human urine samples spiked with varying hCG concentrations (commercially available and traceable to 4th WHO international standard). Provenance is likely laboratory-prepared.
Method Comparison with Predicate Device:
- Sample Size: 100 urine samples for each format (total 300 samples).
- Provenance: Collected from 100 women (about half pregnant, early stage at less than 5 weeks). Samples randomly collected at various times throughout the day. Ages ranged from 20 to 45 years. The provenance is clinical/human samples, seemingly prospective (collected for this study given the blind labeling and specific testing procedure). Country of origin is not specified.
Lay Person Study:
- Sample Size: 100 lay persons for the strip devices, 100 for the cassette devices, and 100 for the midstream devices (total 300 lay persons).
- Provenance: For samples with known hCG concentrations, urine samples were prepared at 5mIU/ml, 8.0mIU/ml, 8.0mIU/ml hCG by spiking hCG into negative pooled urine specimens (laboratory-prepared). Additionally, each participant tested "her own urine sample" (clinical/human, self-collected). Provenance for lay users is diverse educational and professional backgrounds, aged 21 to >50 years, likely within the location of the study (not specified).
False Positive Rate Study:
- Sample Size: 300 non-pregnant females for strip, 300 for cassette, and 300 for midstream (total 900 non-pregnant female tests).
- Provenance: Non-pregnant females who tested "their own urine samples" (clinical/human, self-collected). Provenance likely within the location of the study (not specified).
Early Pregnancy Test Study:
- Sample Size: 585 urine samples from 65 characterized cycle segments of conceptive cycles were collected from 65 pregnant women.
- Provenance: Clinical/human samples from pregnant women.

3. Number of Experts Used to Establish the Ground Truth for the Test Set and Qualifications of Those Experts

Analytical Performance:
- Sensitivity/Cut-Off: 12 operators tested each standard. Qualifications not specified, but likely laboratory technicians given the nature of the testing.
- Specificity & Cross-reactivity: 3 different operators. Qualifications not specified, but likely laboratory technicians.
- Precision: 9 Point-of-Care (POC) operators (3 different operators for each of 3 formats). Qualifications not specified, but suggests healthcare professionals.
Method Comparison with Predicate Device:
- Ground Truth: The "predicate device" serves as the reference standard.
- Testing Personnel: 9 POC operators (three different health professionals at each of the 3 POC sites). Qualifications of "health professionals" not specified but implies skilled personnel.
Lay Person Study:
- Ground Truth: Professional testing of the same samples.
- Testing Personnel: "Professional" testing results are compared to lay person results. The qualifications of these professionals are not explicitly stated, but it is implied they are trained individuals capable of accurately interpreting the test results.
False Positive Rate Study:
- Ground Truth: Derived from professional testing and, in some cases, ultrasound scan confirmation for the false positive results.
- Testing Personnel: The results were from the non-pregnant females themselves, but the ground truth was also established by professional testing and, for the identified false positives, confirmed by ultrasound.
Early Pregnancy Test Study:
- Ground Truth: "Characterized cycle segments of conceptive cycles" from pregnant women indicate clinical diagnosis of pregnancy (likely through follow-up serum hCG, ultrasound, or confirmed delivery/outcome).
- Testing Personnel: Tests were done by an unspecified number of individuals using all three formats. Implicitly, clinical professionals would have characterized the cycle segments.

4. Adjudication Method for the Test Set

The document does not explicitly describe a formal adjudication (e.g., 2+1, 3+1) method for conflicting interpretations of test results within the studies. Observations:

Analytical Performance (Sensitivity/Cut-Off, Interference, Precision): For the sensitivity/cut-off and precision studies, results are reported as aggregates from multiple tests by multiple operators. It appears individual operator results were combined (e.g., 10-/10+ implies 10 negative, 10 positive results out of 20 by various operators). There's no mention of a consensus process for discrepant results at the individual reading level.
Method Comparison and Lay Person Studies: These studies compare the device results to either a predicate device or professional results. The "Professional" results are presented as a single ground truth, implying any potential disagreements among professionals for the ground truth were already resolved or not present.
False Positive Rate Study: For the two cases of false positives, an ultrasound scan was used for confirmation, which serves as a higher-level adjudication for those specific ambiguous cases.

5. If a Multi Reader Multi Case (MRMC) Comparative Effectiveness Study was done, If so, what was the effect size of how much human readers improve with AI vs without AI assistance

An MRMC comparative effectiveness study was not performed in the context of AI assistance. This document describes a medical device (pregnancy test strips/cassettes/midstream) that is a rapid chromatographic immunoassay, not an AI-powered diagnostic tool. Therefore, the concept of "human readers improve with AI vs without AI assistance" is not applicable here.

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

This is not applicable, as the device is a manual, qualitative immunoassay. There is no algorithm or AI component. The user (human-in-the-loop) visually interprets the results.

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

The type of ground truth varied by study:

Analytical Performance:
- Sensitivity/Cut-Off, Specificity, Interfering Substances, Precision, Urine Properties: Laboratory-prepared hCG standards (calibrated against WHO 4th IS for hCG) in negative urine pools, or negative urine samples spiked with known substances. This represents a highly controlled and quantitative ground truth.
Method Comparison with Predicate Device:
- The predicate device (FIRST RESPONSE™ Early Result Pregnancy Test) served as the comparative ground truth.
Lay Person Study:
- Professional testing results of the same samples. For samples with known hCG concentrations, the prepared hCG concentrations served as ground truth.
False Positive Rate Study:
- Confirmed non-pregnancy status (e.g., patient history, and in specific false positive cases, confirmed by ultrasound scan).
Early Pregnancy Test Study:
- Characterized conceptive cycles from pregnant women. This implies clinical confirmation of pregnancy based on biological outcomes (i.e., actual pregnancy).

8. The Sample Size for the Training Set

This document describes performance studies for an existing medical device, not the development or training of an AI algorithm. Therefore, there is no "training set" in the context of machine learning. The studies described are validation and verification studies for the device's performance.

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

As there is no training set for an AI algorithm, this question is not applicable.

Summary

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

GUANGZHOU WONDFO BIOTECH CO., LTD. C/O JOE SHIA BUSINESS DIRECTOR 504 EAST DIAMOND AVE. SUITE F GAITHERSBURG MD 20878

August 7, 2015

Re: K150022

Trade/Device Name: Wondfo One Step HCG Urine Pregnancy Test Strip, Wondfo One Step HCG Urine Pregnancy Test Cassette, Wondfo One Step HCG Urine Pregnancy Test Midstream Regulation Number: 21 CFR 862.1155 Regulation Name: Human chorionic gonadotropin (HCG) test system Regulatory Class: II Product Code: LCX, JHI Dated: March 31, 2015 Received: April 3, 2015

Dear Mr. Joe Shia:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food. Drug. and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA), You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820); and if applicable, the

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electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

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

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

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

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

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

Sincerely yours,

Yung W. Chan -S for

Courtney H. Lias, Ph.D. Director Division of Chemistry and Toxicology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

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Indications for Use

510(k) Number (if known) K150022

Device Name

Wondfo One Step HCG Urine Pregnancy Test Strip Wondfo One Step HCG Urine Pregnancy Test Cassette

Indications for Use (Describe)

Some pregnant women will not be able to detect hCG in their urine 5 days before the expected period. If you test negative before your missed period, but think you may still be pregnant, you should test again a few days after your missed period.

Important note regarding positive results:

Because this test detects low levels of hCG, it is possible that this test may give positive results even if you are not pregnant. If you test positive, but think you may not be pregnant, you should check with your doctor. All results should be confirmed by your healthcare provider, especially when making decisions about future medical care. This product is intended for both prescription use and over-the-counter use.

Important note regarding negative results:

Important note regarding positive results:

All results should be confirmed by your healthcare provider, especially when making decisions about future medical care. This product is intended for both prescription use and over-the-counter use.

Type of Use (Select one or both, as applicable)

X Prescription Use (Part 21 CFR 801 Subpart D)

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

CONTINUE ON A SEPARATE PAGE IF NEEDED.

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Indications for Use

510(k) Number (if known) K150022

Device Name

Wondfo One Step HCG Urine Pregnancy Test Midstream

Indications for Use (Describe)

Important note regarding negative results:

Important note regarding positive results:

Type of Use (Select one or both, as applicable)
-------------------------------------------------	--

| Prescription Use (Part 21 CFR 801 Subpart D)

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

CONTINUE ON A SEPARATE PAGE IF NEEDED.

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510(k) SUMMARY K150022

This summary of 510(k) safety and effectiveness information is being submitted in accordance with the requirements of SMDA 1990 and 21 CFR §807.92.

1. Date:	August 4, 2015
2. Submitter:	Guangzhou Wondfo Biotech Co., Ltd.No.8 Lizhishan Road, Science City,Luogang District, Guangzhou, Guangdong, P.R.China 510663
3. Contact person:	Joe ShiaLSI International Inc.504 East Diamond Ave., Suite FGaithersburg, MD 20878Telephone: 240-505-7880Fax: 301-916-6213Email:shiajl@yahoo.com
4. Device Name:	Wondfo One Step HCG Urine Pregnancy Test StripWondfo One Step HCG Urine Pregnancy Test CassetteWondfo One Step HCG Urine Pregnancy Test Midstream

Classification:

Class II

Product Code	CFR #
LCX, JHI	21 CFR, 862.1155 Human Chorionic Gonadotropin (hCG)Test System

5. Predicate Device:

Church & Dwight Co., Inc. FIRST RESPONSE™ Early Result Pregnancy Test (K123436)

Intended Use 6.

Wondfo One Step HCG Urine Pregnancy Test Strip is a rapid chromatographic immunoassay for the qualitative detection of human chorionic gonadotropin (hCG) in urine, 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.

Important note regarding negative results:

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Important note regarding positive results:

All results should be confirmed by your healthcare provider, especially when making decisions about future medical care.

This product is intended for both prescription use and over-the-counter use.

Important note regarding negative results:

Important note regarding positive results:

All results should be confirmed by your healthcare provider, especially when making decisions about future medical care.

This product is intended for both prescription use and over-the-counter use.

Important note regarding negative results:

Important note regarding positive results:

All results should be confirmed by your healthcare provider, especially when making decisions about future medical care.

This product is intended for over-the-counter use.

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7. Device Description

Similarities and Differences
Item	Candidate device	Predicate device
Intended use	A rapid chromatographicimmunoassay for the qualitativedetection of human chorionicgonadotropin (hCG) in urine, as anaid 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 beforethe day of the missed period.Some pregnant women will not beable to detect hCG in their urine 5days before the expected period.If you test negative before yourmissed period, but think you maystill be pregnant, you should testagain a few days after your missedperiod.Because this test detects low levelsof hCG, it is possible that this testmay give positive results even ifyou are not pregnant. If you testpositive, but think you may not bepregnant, you should check withyour doctor.All results should be confirmed byyour healthcare provider,especially when making decisionsabout future medical care.	An over-the-counter chromatographicimmunoassay for the qualitative detection ofhuman chorionic gonadotropin (hCG) inurine. The device is intended for use as anaid in early detection of pregnancy, in somecases as early as five (5) days before theexpected period, i.e., as early as six (6) daysbefore the day of the missed period.Some pregnant women will not be able todetect hCG in their urine 5 days before theexpected period. If you test negative beforeyour missed period, but think you may stillbe pregnant, you should test again a fewdays after your missed period.Because this test detects low levels of hCG,it is possible that this test may give positiveresults even if you are not pregnant. If youtest positive, but think you may not bepregnant, you should check with yourdoctor.
Specimen	Urine	Same
Assaytechnical	Immunochromatographic assay	Same
Sensitivity	10mIU/ml	Same
Results	Qualitative	Same
Target user	Prescription use( strip and cassette )& over the counter use( strip,cassette and midstream)	Over the counter use
Device format	Strip, cassette, midstream	Midstream
Time to result	5 minutes	3 minutes

8. Substantial Equivalence Information

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Standard/Guidance Document Reference (if applicable)

None

10. Test Principle

It is a lateral flow chromatographic immunoassay. When the absorbent end is immersed into a sample, the sample is absorbed into the device by capillary action and mixes with the antibody-dye conjugate (mouse anti-beta HCG monoclonal antibody), flowing across the pre-coated (Goat anti HCG polyclonal antibody) membrane. During the test procedures, hCG in the urine specimen reacts with the dye conjugate and forms a The complex migrates along the membrane to the a-hCG antibody line (T), and remains complex. captured in the T line. As a result a red colored band develops in the T line, indicating a positive result. If there is no hCG in the urine, there is no red band in the test zone, indicating a negative result. The Control line should develop in the control zone regardless of the test result.

11. Performance Characteristics

A. Analytical performance

a. Sensitivity/ Cut-Off Value
Urine standards containing intact hCG at concentrations of 0, 5, 7.5,8, 9, 10, 15, 20 mIU /ml were prepared in negative urine pool and calibrated against the WHO 4th IS for hCG. Each standard was tested by twelve operators. Each operator tests only one lot one format of the devices of the Wondfo One Step HCG Urine Pregnancy Test. The obtained results are summarized in the following table.

HCGconcentration	Lot I	Lot II	Lot III	%Positive
0mIU/ml	20-/0+	20-/0+	20-/0+	0%
5mIU/ml	20-/0+	20-/0+	20-/0+	0%
7.5mIU/ml	14-/6+	14-/6+	14-/6+	30%
8mIU/ml	10-/10+	10-/10+	10-/10+	50%
9mIU/ml	2-/18+	2-/18+	3-/17+	88.3%
10mIU/ml	0-/20+	0-/20+	0-/20+	100%
15mIU/ml	0-/20+	0-/20+	0-/20+	100%
20mIU/ml	0-/20+	0-/20+	0-/20+	100%

Strip format:

Cassette format:

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HCGconcentration	Lot I	Lot II	Lot III	%Positive
0mIU/ml	20-/0+	20-/0+	20-/0+	0%
5mIU/ml	20-/0+	20-/0+	20-/0+	0%
7.5mIU/ml	14-/6+	14-/6+	15-/5+	28%
8mIU/ml	9-/11+	10-/10+	10-/10+	51.7%
9mIU/ml	2-/18+	2-/18+	3-/17+	88.3%
10mIU/ml	0-/20+	0-/20+	0-/20+	100%
15mIU/ml	0-/20+	0-/20+	0-/20+	100%
20mIU/ml	0-/20+	0-/20+	0-/20+	100%

Midstream format (dip method):

HCGconcentration	Lot I	Lot II	Lot III	%Positive
0mIU/ml	20-/0+	20-/0+	20-/0+	0%
5mIU/ml	20-/0+	20-/0+	20-/0+	0%
7.5mIU/ml	14-/6+	14-/6+	14-/6+	30%
8mIU/ml	9-/11+	10-/10+	10-/10+	51.7%
9mIU/ml	2-/18+	2-/18+	2-/18+	90%
10mIU/ml	0-/20+	0-/20+	0-/20+	100%
15mIU/ml	0-/20+	0-/20+	0-/20+	100%
20mIU/ml	0-/20+	0-/20+	0-/20+	100%

Midstream format (stream method):

HCGconcentration	Lot I	Lot II	Lot III	%Positive
0mIU/ml	20-/0+	20-/0+	20-/0+	0%
5mIU/ml	20-/0+	20-/0+	20-/0+	0%
7.5mIU/ml	15-/5+	14-/6+	14-/6+	28%
8 mIU/ml	9-/11+	10-/10+	10-/10+	51.7%
9 mIU/ml	2-/18+	2-/18+	2-/18+	90%
10mIU/ml	0-/20+	0-/20+	0-/20+	100%
15mIU/ml	0-/20+	0-/20+	0-/20+	100%
20mIU/ml	0-/20+	0-/20+	0-/20+	100%

Cut-off value of 8 mIU/mL and analytical sensitivity of 10 mIU/mL are established.

b. Stability

Stable at 4-30°C for 24 months based on the accelerated stability study at 50°C and real time stability determination at both 4°C and 30°C.

c. Specificity and cross reactivity High Dose Effect

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Negative urine samples were spiked with varying high hCG concentrations ranging from 6,250 to 200,000 mIUmL. The spiked samples were tested by 3 different lots and 3 different operators. No hook effect was observed at these concentrations.

Effects of hCG ß-core fragment

Negative and positive samples (0 and 10 mIU/mL hCG in urine) were spiked with various concentrations of B -core fragment hCG (63,000. 125,000. 250,000 and 500,000 pmol/L). These samples were tested by 3 different lots and 3 different operators. No difference was observed for different lots and different operators. No interference was observed for urine samples with 0mlU/mL hCG and 10mIU/mL hCG for the device except that false negative was observed at the 500000 pmol/L of ß -core fragment hCG.

Effects of glycoprotein LH, FSH and TSH

Negative and positive samples (0 and 10 mIU/mL hCG in urine) were spiked with various concentrations of other glycoprotein hormones such as LH, FSH, and TSH. Samples were tested using three different lots by three operators. No interference was observed for urine samples with 0mIU/mL hCG and 10mIU/mL hCG for the device at LH concentrations up to 500 IU/mL, FSH concentrations up to 1000 mIU/mL, and TSH concentrations up to 1000 µIU/mL

d. Interfering substance

To evaluate potential interference from certain exogenous compounds, each interferent was made at 100X concentrate bulk and spiked in both hCG free and hCG positive (10 and 100mIU/mL) samples. Each spiked urine sample was mixed for 5 minutes to ensure a homogeneous solution before testing. Each sample was tested using 3 different lots of the testing kit. Results are shown in the following table.

Substances tested	Results0mIU/ml hCG			10mIU/ml hCG			100mIU/ml hCG
	Lot 1	Lot 2	Lot 3	Lot 1	Lot 2	Lot 3	Lot 1	Lot 2	Lot 3
Acetaminophen 20 mg/dl	-	-	-	+	+	+	+	+	+
Acetylsalicylic acid 20mg/dl	-	-	-	+	+	+	+	+	+
Ascorbic acid 20mg/dl	-	-	-	+	+	+	+	+	+
Atropine 20mg/dl	-	-	-	+	+	+	+	+	+
Caffeine 20mg/dl	-	-	-	+	+	+	+	+	+
Gentisic acid 20mg/dl	-	-	-	+	+	+	+	+	+
Glucose 2g/dl	-	-	-	+	+	+	+	+	+
Hemoglobin 20mg/dl	-	-	-	+	+	+	+	+	+
Tetracycline 20mg/dl	-	-	-	+	+	+	+	+	+
Ampicillin 20mg/dl	-	-	-	+	+	+	+	+	+
Albumin 20mg/dl	-	-	-	+	+	+	+	+	+
B-hydroxybutyrate	-	-	-	+	+	+	+	+	+

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(2000mg/dL)
Ephedrine (20mg/dL)	-	-	-	+	+	+	+	+	+
Phenylpropanolamine(20mg/dL)	-	-	-	+	+	+	+	+	+
Phenothiazine (20mg/dL)	-	-	-	+	+	+	+	+	+
EDTA (80mg/dL)	-	-	-	+	+	+	+	+	+
Salicyclic Acid (20mg/dL)	-	-	-	+	+	+	+	+	+
Benzoylecgonine (10mg/dL)	-	-	-	+	+	+	+	+	+
Cannabinol (10mg/dL)	-	-	-	+	+	+	+	+	+
Codeine (6ug/dL)	-	-	-	+	+	+	+	+	+
Ethanol (1.0%)	-	-	-	+	+	+	+	+	+
Bilirubin (2mg/dL)	-	-	-	+	+	+	+	+	+
Atropine (20mg/dL)	-	-	-	+	+	+	+	+	+
Pregnanediol (1500ug/dL)	-	-	-	+	+	+	+	+	+
Thiophene (20mg/dl)	-	-	-	+	+	+	+	+	+
Ketone(20mg/dl)	-	-	-	+	+	+	+	+	+

All data show that there is no interference for the listed compounds at the stated concentrations.

e. Effect of Urine Specified Gravity and Urine pH

Negative and positive urine samples containing 0, 10 and 100 mIU/mL hCG were tested at pH values from 4 to 9 or at density values ranging from 1.000 to1.035 using 3 different operators. Data show that there is no interference from pH ranging from 4 to 9 and specific gravity ranging from 1.000 to 1.035 of tested urine samples

f. Precision

A precision study was performed using negative human urine samples spiked with varying hCG (commercially available and traceable to the 4"WHO international standard) concentrations. The spiked urine samples were measured in 10 replicates per day using 3 different lots for each format. Tests were performed for 5 consecutive days by three different operators at each of 3 point-of-care (POC) sites for a total of 9 POC operators. Different set of operators tested each format. Results are shown in the following tables.

hCG Concentration(mIU/ml)	Lot 1	Lot 2	Lot 3
0	50-/0+	50-/0+	50-/0+
2.5	50-/0+	50-/0+	50-/0+
5	50-/0+	50-/0+	50-/0+
7.5	36-/14+	35-/15+	36-/14+
8	24-/26+	25-/25+	24-/26+
10	0-/50+	0-/50+	0-/50+

Strip format:

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15	0-/50+	0-/50+	0-/50+
20	0-/50+	0-/50+	0-/50+
25	0-/50+	0-/50+	0-/50+
50	0-/50+	0-/50+	0-/50+

Cassette format:

hCG Concentration(mIU/ml)	Lot 1	Lot 2	Lot 3
0	50-/0+	50-/0+	50-/0+
2.5	50-/0+	50-/0+	50-/0+
5	50-/0+	50-/0+	50-/0+
7.5	35-/15+	35-/15+	36-/14+
8	25-/25+	24-/26+	24-/26+
10	0-/50+	0-/50+	0-/50+
15	0-/50+	0-/50+	0-/50+
20	0-/50+	0-/50+	0-/50+
25	0-/50+	0-/50+	0-/50+
50	0-/50+	0-/50+	0-/50+

Midstream format:

hCG Concentration(mIU/ml)	Lot 1	Lot 2	Lot 3
0	50-/0+	50-/0+	50-/0+
2.5	50-/0+	50-/0+	50-/0+
5	50-/0+	50-/0+	50-/0+
7.5	35-/15+	36-/14+	36-/14+
8	24-/26+	24-/26+	24-/26+
10	0-/50+	0-/50+	0-/50+
15	0-/50+	0-/50+	0-/50+
20	0-/50+	0-/50+	0-/50+
25	0-/50+	0-/50+	0-/50+
50	0-/50+	0-/50+	0-/50+

B. Comparison study

Method comparison with predicate device

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A method comparison study was performed, comparing the results obtained from the Wondfo One Step HCG Urine Pregnancy Test to the results from predicate devices. 100 urine samples were collected for each format from 100 women (about half of them were pregnant, early stage at less than 5 weeks) for a total of 300 samples. Samples were randomly collected at various times throughout the day. Ages ranged from 20 to 45 years. The samples were blind labeled. Samples were tested by three different health professionals at each of the 3 POC sites for a total of 9 POC operatorswith the proposed and the predicate devices. Each person could only perform tests for one device format. For example, a person who tested the strips could not test the cassettes or the midstream. Each person tested three different lots of the device and one predicate device at the same time, but not sequentially. Summary results are shown in the following tables.

For Strip Format

Wondfo DeviceViewer A, B, CLot 1, 2, 3	Predicate device	+	-
+		49	0
-		0	51

For Cassette Format

Wondfo DeviceViewer D, E, FLot 1, 2, 3	Predicate device	+	-
+		48	0
-		0	52

For Midstream Format

Wondfo DeviceViewer G, H, ILot 1, 2, 3	Predicate device	+	-
+		49	0
-		0	51

Results show that 100% agreements for both positive and negative samples.

Lay person study

A lay user study was performed at three intended use sites with 100 lay persons testing the strip devices, 100 lay persons testing the cassette devices and another set of 100 persons testing the midstream devices (60 by stream method and 40 by dip method). A total of 300 females with diverse educational and professional backgrounds and ranged in age from 21 to > 50 years performed the study. Urine samples were prepared at 5mIU/ml, 8.0mIU/ml, 8.0mIU/ml hCG concentrations by spiking hCG into negative pooled urine specimens. Each sample was aliquoted into individual containers and blind-labeled. Each participant was provided with the package insert, 2 blind labeled samples (5mlU/ml and 10mlU/ml) and Wondfo's test kits. Each lay person also tested her own urine sample using the Wondfo device and provided a sample for professional testing. Additional lay-person tests were performed for the spiked hCG 7.5mIU/ml and 8.0mIU/ml samples by 100 lay persons for each device formats.

The results are summarized below.

Strip format

	Professional	+	-
Lay person	+	49	0
	-	0	51

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Numberofsamples	hCG Concentration(mIU/ml)	Lay person results		The percentage ofcorrect results (%)
100	5	No. ofPositive	No. ofNegative	99%
100	10	1	99	100%

Number	hCG Concentration(mIU/mL)	Lay person results		Professionals results		Consistencerate(%)
ofsamples			No. ofPositive	No. ofNegative	No. ofPositive	No. ofNegative
100	7.5	26	74	30	70	96%
100	8	49	51	52	48	97%

Cassette format

	Professional	+	-
Lay person	+	48	1
	-	0	51

Numberofsamples	hCG Concentration(mIU/ml)	Lay person results		The percentage ofcorrect results (%)
		No. ofPositive	No. ofNegative
100	5	0	100	100%
100	10	100	0	100%

Numberofsamples	hCG Concentration(mIU/mL)	Lay person results		Professionals results		Consistencerate(%)
100	7.5	25	75	29	71	96%
100	8	47	53	51	49	96%

Midstream format

	Professional	+	-
Lay person	+	49	0
	-	0	51

Numberofsamples	hCG Concentration(mIU/ml)	Lay person results		The percentage ofcorrect results (%)
		No. ofPositive	No. ofNegative
100	5	0	100	100%
100	10	99	1	99%

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Numberofsamples	hCG Concentration(mIU/mL)	Lay person results		Professionals results		Consistencerate(%)
100	7.5	25	75	29	71	96%
100	8	49	51	52	48	97%

Lay-users were also given surveys on the ease of understanding the package insert instructions. All lay users indicated that the device instructions can be easily followed. A Flesch-Kincaid reading analysis was performed on each package insert and the scores revealed a reading Grade Level of 7.

False Positive Rate Study

In this study, 300 non-pregnant females tested their own urine samples using the strip devices. The obtained results show that the test strip yields 0.3% false positive results.

LotGroup	I	II	III
Pre-menopasual	0+/34-	0+/33-	0+/33-
Peri-menopasual	1+*/32-	0+/34-	0+/33-
Post-menopasual	0+/33-	0+/33-	0+/34-

*This result was from a 49 year old woman. The ultrasound scan showed non- pregnancy.

Another 300 non-pregnant females tested their own urine samples using the cassette devices. The obtained results show that the test cassette yields 0.3% false positive results.

LotGroup	I	II	III
Pre-menopasual	0+/34-	0+/33-	0+/33-
Peri-menopasual	0+/33-	0+/34-	1+*/32-
Post-menopasual	0+/33-	0+/33-	0+/34-

*This result was from a 51 years old woman. The ultrasound scan showed non- pregnancy.

The last 300 non-pregnant females tested their own urine samples using the midstream devices. The obtained results show none false positive results.

LotGroup	I	II	III
Pre-menopasual	0+/34-	0+/33-	0+/33-
Peri-menopasual	0+/33-	0+/34-	0+/33-
Post-menopasual	0+/33-	0+/33-	0+/34-

Early Pregnancy Test Study

In this study, total 585 urine samples from 65 characterized cycle segments of conceptive cycles were collected from 65 pregnant women. All samples were masked and randomized. Each sample was tested by all three formats of the device. The new device detected 68% positive hCG five days before the Expected Menstrual Period (EMP), and 100% positive hCG on the day of EMP. No differences were observed between different device formats. The following table is the summary of the data.

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Day relative to EMP	EMP-8	EMP-7	EMP-6	EMP-5	EMP-4	EMP-3	EMP-2	EMP-1	EMP
# of cycles positive for hCG	4/65	9/65	25/65	44/65	58/65	63/65	64/65	65/65	65/65
% cycles positive for hCG	6%	14%	38%	68%	89%	97%	98%	100%	100%

12. Conclusion

Based on the test principle and acceptable performance characteristics including precision, cut-off, interference, specificity and method comparison of the devices, it's concluded that Wondfo One Step HCG Urine 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.