Macro & Micro-Test HCG Pregnancy Test Midstream is intended for over-the-counter use for qualitative detection of hCG hormone in urine to aid early detection of pregnancy.

Device Description

Macro & Micro-Test HCG Pregnancy Test Midstream is a qualitative lateral flow test for the detection of hCG (human chorionic gonadotropin) hormone, which is present in urine during the early stage of pregnancy. The midstream kits consist of one test device and a package insert. The test device contains mouse monoclonal anti- B hCG antibody coated membrane and a pad containing mouse monoclonal anti- a -hCG antibody colloidal gold conjugate. The control antibodies are goat anti-rabbit IgG.

AI/ML Overview

The provided document is a 510(k) summary for the Macro & Micro-Test HCG Pregnancy Test Midstream. It contains information about the device's acceptance criteria and the studies performed to demonstrate its performance.

Here's the requested information:

1. Table of Acceptance Criteria and Reported Device Performance:

Acceptance Criteria	Reported Device Performance
LoD/Sensitivity (Detection Limit)	25 mIU/mL: The results indicate that the positive detection rate is more than 95% when the concentration is not less than 25 mIU/mL.
Consistency/Agreement with Predicate	100% agreement (Positive Percent Agreement, Negative Percent Agreement, Total Percent Agreement) with the predicate device (midstream version) for 250 urine samples.
Lay User Agreement with Professionals	100% agreement between lay user results and professional results for 250 urine samples. Lay users also found the product instructions easy to understand and devices easy to use.
Precision/Reproducibility	Consistent and precise test results across 3 batches, 3 sites, 5 days, and 10 replicates per day for various hCG concentrations (0, 12.5, 18.75, 25, 50, 100 mIU/mL). Example: For 25 mIU/mL, 100% positive across all sites; for 0 and 12.5 mIU/mL, 100% negative across all sites. At 18.75 mIU/mL, 53.33% positive and 46.67% negative, reflecting expected near-LoD performance.
Specificity (Cross-reactivity)	No cross-reaction observed with hLH at 500mIU/ml, hFSH at 1000 mIU/mL, and hTSH at 1000 µIU/mL.
Interfering Substances	No interference observed from 29 common exogenous compounds (e.g., Acetaminophen, Caffeine, Hemoglobin, Vitamin C, Glucose, Bilirubin, etc.) at specified concentrations in both positive (25mIU/mL hCG) and negative urine samples.
Effects of Urine pH	No interference observed for urine pH range of 4-9 (tested with 0 and 25mIU/mL hCG).
Effects of Urine Specific Gravity	No interference observed for urine with specific gravity 1.000-1.035 (tested with 0 and 25mIU/mL hCG).
High Dose Hook Effect	No high dose hook effect for hCG concentrations up to 1,000,000 mIU/mL (Note: The study in section H.h only tested up to 1000IU/ml, which is 1,000,000 mIU/mL. The table in G indicates a claim of no hook effect up to 1,000,000 mIU/mL).
Beta hCG Core Fragment Reactivity	No interference observed when the concentration of HCG ß-core fragment reaches 1 µmol/L.
Read Time	Optimal interpretation within 5-10 minutes. Test result was "most accurate" when interpreted within this window (tested at 5, 15, 20, and 30 minutes).
Shelf Life	24 months when stored at 4-30°C.
Quality Control	Appearance of a red band at the control line (C) and a clear background in the results window.

2. Sample Sizes Used for the Test Set and the Data Provenance:

Consistency Study (Comparison with Predicate): 250 urine samples.
- Data Provenance: Not explicitly stated (e.g., country of origin). The document mentions "Urine samples were selected from 250 women aged from 18-55 years old." It doesn't specify if the study was retrospective or prospective. Given the testing methodology described, it appears to be a prospective collection or at least a prospective testing of collected samples.
OTC Lay User Test: 250 urine samples.
- Data Provenance: Not explicitly stated (e.g., country of origin). The study involved "women aged from 18-55 years old". It seems to be a prospective study, as participants were given the device to test samples and questionnaires were administered.
Precision/Reproducibility Study: 900 urine samples (across 6 concentrations: 0, 12.5, 18.75, 25, 50, 100 mIU/mL; 3 sites x 5 days x 10 replicates per concentration).
- Data Provenance: Not explicitly stated, but conducted at "3 sites (one internal site and two external sites representative of the intended use setting)." This suggests the study was conducted within a controlled setting.
LoD/Sensitivity Study: 30 negative samples used to dilute WHO International Standard hCG to various concentrations (0, 12.5, 18.75, 25, 50, and 100 mIU/mL). The number of test replicates at each concentration for LoD determination is not explicitly stated, but the precision study data for similar concentrations would provide insight into reproducibility near the LoD.
- Data Provenance: Not explicitly stated.

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

Consistency Study: Test results were compared between the candidate device and the predicate device. For the candidate device, testing was done by "three different health professionals at each of three sites for a total of three operators." The predicate device results would likely also have been generated by trained professionals.
- Qualifications: "Health professionals" are mentioned, but specific qualifications (e.g., "Radiologist with 10 years of experience") are not provided.
OTC Lay User Test: "The same urine samples were tested by professionals in parallel using the proposed device." These professional results served as the ground truth against which lay user results were compared.
- Qualifications: "Professionals" are mentioned, but specific qualifications are not detailed beyond that.
Analytical Studies (Specificity, Interfering Substances, pH, Specific Gravity, High Dose Hook, Beta hCG Core Fragment Reactivity, LoD, Read Time, Precision): The ground truth for these studies was established by preparing urine samples with known concentrations of hCG, interferents, or specific environmental conditions, traceable to international standards (e.g., WHO 6th IS for hCG). This relies on laboratory standards and scientific measurement rather than expert interpretation of results.

4. Adjudication Method for the Test Set:

Consistency Study & OTC Lay User Test: No explicit detailed adjudication method (like 2+1, 3+1) is described. The agreement was calculated based on direct comparison of results. For the consistency study, the "agreement of hCG pregnancy test midstream with the predicate device was 100%," implying direct concordance. For the lay user study, "100% agreement between lay user results and professional results" also indicates direct concordance. This suggests a direct comparison method.
Analytical Studies: Adjudication is not typically applicable for these types of controlled analytical tests where known quantities are used.

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:

No, an MRMC comparative effectiveness study was not done. This document pertains to an in vitro diagnostic (IVD) device (pregnancy test midstream) rather than an AI-powered diagnostic imaging device. Therefore, the concept of "human readers improve with AI vs without AI assistance" does not apply here.

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

This device is a standalone test kit (a lateral flow immunoassay), not an algorithm or an AI-based system. Its performance is inherent to the chemical reactions and visual interpretation, which can be done by a human (lay user or professional). The term "standalone" in the context of AI algorithms is not directly applicable here. The device itself is the "standalone" diagnostic tool. The document does report its performance when used by professionals (which can be considered the equivalent of "algorithm only" in this context, as professionals are interpreting the device's output) and by lay users.

7. The type of ground truth used:

For consistency and lay user studies: The results from the predicate device (or professional testing in the case of the lay user study) served as a comparative ground truth.
For analytical performance studies (LoD, Precision, Specificity, Interfering Substances, pH, Specific Gravity, High Dose Hook, Beta hCG Core Fragment Reactivity): Laboratory-prepared samples with known concentrations of analytes (hCG) or interferents, traceable to international standards (e.g., WHO 6th IS for hCG), were used to establish the ground truth. This is a highly controlled and quantitative ground truth.

8. The sample size for the training set:

This document describes performance studies for a traditional in vitro diagnostic (IVD) lateral flow immunoassay (not an AI/ML device). Therefore, there is no concept of a "training set" in the context of machine learning model development. The samples tested in the studies described are for performance evaluation and validation.

9. How the ground truth for the training set was established:

As stated above, this is not an AI/ML device, so there is no training set and thus no ground truth established for a training set in this context.

Summary

{0}------------------------------------------------

Image /page/0/Picture/0 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".

Jiangsu Macro & Micro-Test Med-Tech Co., Ltd. % Sandra Jiang Shanghai Sungo Management Consulting Company Limited Room 1401, Dongfang Building, 1500# Century Avenue Shanghai. 200122 China

Re: K230741

Trade/Device Name: Macro & Micro-Test HCG Pregnancy Test Midstream Regulation Number: 21 CFR 862.1155 Regulation Name: Human Chorionic Gonadotropin (HCG) Test System Regulatory Class: Class II Product Code: LCX Dated: August 31, 2023 Received: August 31, 2023

Dear Sandra Jiang:

We have reviewed your section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (the Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. Although this letter refers to your product as a device, please be aware that some cleared products may instead be combination products. The 510(k) Premarket Notification Database available at https://www.accessdata.fda.gov/scripts/cdrh/cfdocs/cfpmn/pmn.cfm identifies combination product submissions. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

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

Additional information about changes that may require a new premarket notification are provided in the FDA guidance documents entitled "Deciding When to Submit a 510(k) for a Change to an Existing Device" (https://www.fda.gov/media/99812/download) and "Deciding When to Submit a 510(k) for a Software Change to an Existing Device" (https://www.fda.gov/media/99785/download).

{1}------------------------------------------------

Your device is also subject to, among other requirements, the Quality System (OS) regulation (21 CFR Part 820), which includes, but is not limited to, 21 CFR 820.30, Design controls; 21 CFR 820.90, Nonconforming product; and 21 CFR 820.100, Corrective and preventive action. Please note that regardless of whether a change requires premarket review, the QS regulation requires device manufacturers to review and approve changes to device design and production (21 CFR 820.30 and 21 CFR 820.70) and document changes and approvals in the device master record (21 CFR 820.181).

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR Part 803) for devices or postmarketing safety reporting (21 CFR Part 4, Subpart B) for combination products (see https://www.fda.gov/combination-products/guidance-regulatory-information/postmarketing-safetyreporting-combination-products); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR Part 820) for devices or current good manufacturing practices (21 CFR Part 4, Subpart A) for combination products; and, if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR Parts 1000-1050.

Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to https://www.fda.gov/medical-device-safety/medical-device-reportingmdr-how-report-medical-device-problems.

For comprehensive regulatory information about mediation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/medicaldevices/device-advice-comprehensive-regulatory-assistance) and CDRH Learn (https://www.fda.gov/training-and-continuing-education/cdrh-learn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (https://www.fda.gov/medical-device-advice-comprehensive-regulatoryassistance/contact-us-division-industry-and-consumer-education-dice) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).

Sincerelv. Joseph A. Digitally signed by Joseph A. Kotarek -S Kotarek -S Date: 2023.10.13 Joseph Kotarek 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

{2}------------------------------------------------

Indications for Use

510(k) Number (if known) K230741

Device Name

Macro & Micro-Test HCG Pregnancy Test Midstream

Indications for Use (Describe)

Macro & Micro-Test HCG Pregnancy Test Midstream is intended for over-the-counter use for qualitative detection of hCG hormone in urine to aid early detection of pregnancy.

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.

This section applies only to requirements of the Paperwork Reduction Act of 1995.

DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.

The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:

Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff@fda.hhs.gov

"An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number."

{3}------------------------------------------------

510(K) Summary

K230741

(As requirement by 21 CFR 807.92)

Date prepared: 17th, Aug 2023

A. Applicant:

Name: Jiangsu Macro & Micro-Test Med-Tech Co., Ltd. Address: No.888, Zhujiang Road, Juegang Street, Rudong County (Life & Health of Rudong High-tech Zone) Nantong City, Jiangsu 226499, China Contact Person: Bella Xu Title: Regulatory Affairs Tel: +86 15195862818 Email: xuyufeng@hongweitest.com

Submission Correspondent: Primary contact: Sandra Jiang Shanghai SUNGO Management Consulting Co., Ltd. Room 1401, Dongfang Building, 1500# Century Ave., Shanghai 200122, China Tel: +86-21-58817802 Email: yuan.jiang(@sungoglobal.com Secondary contact: Mr. Raymond Luo Room 1401, Dongfang Building, 1500# Century Ave., Shanghai 200122, China Tel: +86-21-68828050 Email: fda.sungo@@gmail.com

B. Device:

Trade Name: Macro & Micro-Test HCG Pregnancy Test Midstream

Regulatory Information Classification Name: Kit, Test, Pregnancy, Hcg, Over The Counter Classification: Class II Product code: LCX Regulation Number: 21 CFR 862.1155 Review Panel: Clinical Chemistry

C. Predicate device:

K150063

ACCU NEWS One Step hCG Pregnancy Test Strip

ACCU NEWS One Step hCG Pregnancy Test Cassette

{4}------------------------------------------------

ACCU NEWS One Step hCG Pregnancy Test Midstream

D. Indications for use of the device:

Macro & Micro-Test HCG Pregnancy Test Midstream is intended for over-the-counter use for qualitative detection of hCG hormone in urine to aid early detection of pregnancy.

E. Device Description:

The Macro & Micro-Test HCG Pregnancy Test Midstream will be sold in one format: midstream. The midstream kits consist of one test device and a package insert. The test device contains mouse monoclonal anti- B hCG antibody coated membrane and a pad containing mouse monoclonal anti- a -hCG antibody colloidal gold conjugate. The control antibodies are goat anti-rabbit IgG.

F. Principle of Operation

The product uses the reaction principle of double antibody sandwich method and is developed in combination with immunochromatography technology. The goat anti-mouse polyclonal antibody is coated at the quality control line (C) on the nitrocellulose membrane, and the HCG monoclonal antibody is coated at the test line (T) on the nitrocellulose membrane. The HCG monoclonal antibody labeled with red tracers is coated on the glass cellulose membrane. During detection, the sample flows upward under the action of chromatography, and when the HCG concentration in the sample is higher than the LoD of this test strips, HCG will specifically bind to the anti-HCG and be captured by the HCG monoclonal antibody at the test line, thus a red band appears at the test line. When the HCG concentration in the sample is lower than the LoD of this test strip or there is no HCG in the sample, then no red band appears at the test line. Regardless of the content of HCG in the samples, anti-HCG can be captured by the goat anti-mouse polyclonal antibody at the quality control line to form an immune complex, thus a red band appears at the quality control line.

Device	Proposed Device	Predicate Device	Result
510K #	K230741	K150063	-
Basic Information Comparison
Manufacturer	Jiangsu Macro & Micro-TestMed-Tech Co., Ltd.	Coretests Inc.	-
Intended use	It is intended for qualitative detectionof hCG hormone in urine to aid earlydetection of pregnancy.	It is intended for qualitative detection ofhCG hormone in urine to aid earlydetection of pregnancy.	Same
Intended useenvironment	Over the Counter (OTC)	Over the Counter (OTC) andprofessional	Different

G. Summary of Technological Characteristics

{5}------------------------------------------------

Specimen Type	Urine	Urine	Same
Technology	Lateral Flow Immunoassay	Lateral Flow Immunoassay	Same
BasicComponents	Test strip containing nitrocellulosemembrane, sample pad, alpha andbeta hCG antibodies	Test strip containing nitrocellulosemembrane, sample pad, alpha and betahCG antibodies	Same
StorageTemperature	4-30°C	4-30°C	Same
Shelf life	24 months when stored at 4-30°C	24 months when stored at 4-30°C	Same
Format	Midstream	Strip, Cassette, Midstream	Same
Read Time	At 5 minutes	At 5 minutes	Same
Result	Qualitative	Qualitative	Same
Analytical Performance Comparison
Detection Limit	25 mIU/mL	25 mIU/mL	Same
High DosageHook Effect	No high dosage hook effect forhCG up to 1,000,000 mIU/mL	No high dosage hook effect forhCG up to 500,000 mIU/mL	Similar
Specificity	LH at 500mIU/mL, FSH at 1000mIU/mL, and TSH at 1000 µIU/mL	LH at 1000mIU/mL, FSH at 1000mIU/mL, and TSH at 1000 µIU/mL	Similar
pHInterference	No interference for urine with pH 4-9	No interference for urine with pH 2-9	Similar
SpecificGravityInterference	No interference for urine withSpecific Gravity 1.000-1.035	No interference for urine withSpecific Gravity 1.000-1.030	Similar
Beta hCG CoreFragmentReactivity	No interference when theconcentration of HCG β-corefragment reaches 1µmol/L.	No interference when the concentrationof HCG β-core fragment reaches 1µmol/L.	Same

Analysis:

The proposed device is substantially equivalent to the predicate device (midstream version), in terms of intended use, format, test principle, shelf life, read time and LoD.

Although the indications of the predicate device and the proposed device are different, i.e. OTC and professional for predicate device and OTC for proposed device, the indication of the proposed device is covered by that of the predicate device. Such difference will not give any impact on the safety and effectiveness of the proposed deice.

The minor difference of analytical performance between the predicate device (midstream version) and the proposed device will not influence the effectiveness of the proposed device.

Therefore, it can be concluded that the subject device and the proposed device are substantially equivalent.

H. Summary of Non-Clinical Testing

a. Comparison Studies

Consistency of Proposed device and the Predicate Device (midstream version)

{6}------------------------------------------------

Urine samples were selected from 250 women aged from 18-55 years old. About half of the selected women were suspected to be pregnant (less than 5 weeks pregnant, including women who tested as early as the day of their missed period). The samples were tested by three different health professionals at each of three sites for a total of three operators with the candidate device and the midstream version of predicate device. Test results were compared. The data show that the agreement of hCG pregnancy test midstream with the predicate device was 100%.

		Predicate Device
		Positive (+)	Negative (-)	Total
CandidateDevice	Positive	127	0	127
	Negative	0	123	123
	Total	127	123	250
EvaluationIndex	Kappa Value	Positive PercentAgreement	Negative PercentAgreement	Total PercentAgreement
	1	100%	100%	100%

Table 1. Test Results of 250 samples of Test Midstream

● OTC Layer User Test Results

Urine samples were selected from 250 women aged from 18-55 years old. About half of the selected women were suspected to be pregnant (less than 5 weeks pregnant, including women who tested as early as the day of their missed period). Those women were given Macro & Micro-Test HCG Pregnancy Test Midstream to test the collected urine samples. The same urine samples were tested by professionals in parallel using the proposed device. The recorded test results from the lay users were compared with those from professionals. In parallel, after testing, each participant was given a questionnaire to assess the readability of the instructions for use. The result showed that the lay users found the product instructions were easy to understand and the devices were easy to use.

		Professionals Test Results
		Positive (+)	Negative (-)	Total
Lay User TestResults	Positive	127	0	127
	Negative	0	123	123
	Total	127	123	250
	% Agreement	100%

The test result showed that 100% agreement between lay user results and professional results.

{7}------------------------------------------------

b. Read Time

To determine the optimal time to interpret the test results of the hCG pregnancy test midstream. 10 non-pregnant samples were selected. Add HCG standard to 3 samples to obtain the HCG positive samples with concentration of 25mIU/mL, 100mIU/mL, After the test is completed, interpret the results after 5 minutes, 15 minutes, 20 minutes, and 30 minutes. Data indicated that test result was most accurate when it was interpreted within 5-10 minutes.

c. Precision/Reproducibility

Three batches of device were used to test urine samples containing 0, 12.5, 18.75, 25, 50, and 100mIUmL of hCG. The Urine samples were prepared by using hCG free urine samples spiked with hCG to achieve the following concentrations: 0, 12.5, 18.75, 25, 50, and 100 mIU/mL. All urine samples were traceable to the hCG standard WHO 6th IS. The study was carried out at 3 sites (one internal site and two external sites representative of the intended use setting). Each site tested a unique lot of test device. Each sample was tested in 10 replicates per day. Repeat the test for 5 non-consecutive days for each device lot, 150 Urine samples were tested for each concentration ( 3sites(lots) ×5days ×10 replicates). Totally, 900 urine samples × 6 concentrations ) were tested.

hCGConcentration(mIU/mL)	2105MY19001(Location 1)		2105MY24001(Location 2)		2105MY27001(Location 3)		%Positive	%Negative
	+	-	+	-	+	-
0	0	50	0	50	0	50	100%	0%
12.5	0	50	0	50	0	50	100%	0%
18.75	26	24	26	24	28	22	53.33%	46.67%
25	50	0	50	0	50	0	0%	100%
50	50	0	50	0	50	0	0%	100%
100	50	0	50	0	50	0	0%	100%

Table 3 Precision Study Data for Midtream

Test result demonstrated that hCG pregnancy test midstream was able to produce consistent and precise test results.

d. Specificity

To determine if the hCG pregnancy test midstream was affected (i.e., cross reacted) by a relevant challenge dose of closely related human luteinizing hormone (hLH), human follicle stimulating hormone (hFSH) or human thyroid stimulating hormone (hTSH). The results demonstrated no cross reaction with hLH at 500mIU/ml, hFSH at 1000 mIU/mL, and hTSH at 1000 µIU/mL.

e. Interfering Substances

{8}------------------------------------------------

Jiangsu Macro & Micro-Test Med-Tech Co., Ltd.

No.888, Zhujiang Road, Juegang Street, Rudong County (Life & Health of Rudong High-tech Zone) Nantong City, Jiangsu 226499, China

To determine if the hCG pregnancy test midstream was affected by potentially interfering substances which may be found in human urine, positive urine samples (hCG 25mIU/mL) and negative urine samples were spiked with the interference substance to obtain the specific concentrations. Each spiked urine sample was tested with the results show that no interference was observed from exogenous compounds at the different concentrations for both negative and positive HCG urine samples.

Interfering Substances	Final Concentration	Interfering Substances	Final Concentration
Acetaminophen	20mg/dL	Hemoglobin	1000mg/dL
ẞ-hydroxybutyrate	2000mg/dL	Pregnanediol	1500µg/dL
Caffeine	20mg/dL	Thiophene	20mg/dL
Phenylpropanolamine	20mg/dL	Ampicillin	20mg/dL
Salicylic acid	20mg/dL	Tetracycline	20mg/dL
Phenothiazine	20mg/dL	Vitamin C	20mg/dL
Aspirin	80mg/dL	(1R,2S)-(-)-Ephedrine	20 mg/dL
Cannabinol	10mg/dL	(1S,2R)-(+)-Ephedrine	20 mg/dL
Ethanol	1.0%	Acetone	20 mg/dL
Methanol	10%	Gentisic Acid	20 mg/dL
Albumin	2000mg/dL	Ibuprofen	40 mg/dL
Glucose	2000mg/dL	Methadone	20 mg/dL
Bilirubin	40mg/dL	Morphine	10 mg/dL
Atropine	20mg/dL	Nicotine	20 mg/dL
Estriol-17-β	1400µg/dL	Uric acid	20 mg/dL
Amoxicillin	20mg/dL	Benzoylecgonine	10 mg/dL
Folic acid	0.03 mg/dL	Vitamin B1	80 mg/dL
EDTA	80 mg/dL	Ketone	20 mg/dL

f. Effects of Urine pH

The PH of an aliquot negative urine pool is adjusted to a PH range of 4 to 9 in 1 PH unit increment and spiked with HCG at 25mIU/ml and 0mIU/ml and 3 batches of hCG pregnancy test midstream were tested repeatedly. The result demonstrated that varying ranged of PH do not interfere with the performance of the test.

g. Effects of Urine Specific Gravity

{9}------------------------------------------------

Jiangsu Macro & Micro-Test Med-Tech Co., Ltd.

No.888, Zhujiang Road, Juegang Street, Rudong County (Life & Health of Rudong High-tech Zone) Nantong City, Jiangsu 226499, China

Urine samples containing 0 and 25 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. 3 batches of hCG pregnancy test midstream were tested. The data show that there's no interference in the test result when the specific gravity is between 1.000-1.035.

h. High Dose Hook Effect

The test was evaluated for high dose hook effect. HCG free specimens spiked with the HCG at different concentration containing 0.5IU/ml, 10U/ml, 100IU/ml, 500IU/ml, 1000IU/ml, Three batches of tests were tested.

The result show that hCG pregnancy test midstream can get the positive result when the HCG concentration is range from 0.5IU/ml to 1000IU/ml.

i. Beta hCG Core Fragment Reactivity testing

To evaluate the effects of the HCG ß-core fragment nonpregnant females' urine specimens containing 0 and 25mIU/ml, HCG were spiked with the HCG ß-core fragment at the concentration of 0.5 umol/L. The data shows that there's no interference in the test result when the concentration of HCG ß-core fragment reaches 1 umol/L.

j. LoD/Sensitivity

To determine the LoD of hCG pregnancy test midstream, 30 negative samples were selected to dilute the WHO International Standard 6th International Standard for Chorionic Gonadotrophin (Batch No.: 6th INTERNATIONAL STANDARD 2820) for human chorionic gonadotropin immunoassay to 0, 12.5, 18.75, 25, 50 and 100 mIU/mL . Use the kit for detect the concentration with a positive detection rate of more than 95% as the LoD of the kit.

The results indicate that the positive detection rate is more than 95% when the concentration is not less than 25mIU/mL, so, the LoD of the kit is 25mIU/mL.

k. Expected Value

Not applicable.

l. Calibration

The tests are calibrated against WHO International Standard 6th International Standard.

m. Quality Control

The membrane is pre-coated with anti-mouse antibodies on the control band region. During the testing, the mixture continues to migrate across the membrane to the immobilized goat anti-mouse region, a red band at the control band region will always appear. The presence of this red band

{10}------------------------------------------------

serves as verification for sufficient sample volume and proper flow and as a control for the reagents. The Control line should develop in the control zone regardless of the test result. A red line appearing in the control region (C) is considered an internal positive procedural control. A clear background in the results window in considered an internal negative procedural control. If the Control (C) line does not give the expected reaction on any sample, the test is invalid, and must be repeated.

n. Stability/Shelf Life

The stability data supports that the products have the shelf life of 24 months when stored at 4-30°C.

o. Antibody Information

Antibody	The biological source	The location	Specific targetantigen
Monoclonal anti-β-HCG antibody	Mouse	Conjugate pad	Specific to the beta subunit of hCG
Monoclonal anti-α-H-CG antibody	Mouse	Nitrocellulose membrane	Specific to the alpha subunit of hCG
goat anti mouse lgG polyclonal antibody	Goat	Nitrocellulose membrane

Clinical Test Conclusion I.

No clinical study is included in this submission.

J. Conclusion

The performance characteristics studies performed demonstrated substantial equivalency between the proposed device and predicate device (K150063 ACCU NEWSTM One Step hCG Pregnancy Test -Midstream version).

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.