ONE STEP HCG TEST STRIP, ONE STEP HCG TEST CASSETTE, ONE STEP HCG TEST MIDSTREAM

K132085 · Co-Innovation Biotech Co., Ltd. · LCX · Nov 12, 2013 · Clinical Chemistry

Device Facts

• Record ID: K132085
• Device Name: ONE STEP HCG TEST STRIP, ONE STEP HCG TEST CASSETTE, ONE STEP HCG TEST MIDSTREAM
• Applicant: Co-Innovation Biotech Co., Ltd.
• Product Code: LCX · Clinical Chemistry
• Decision Date: Nov 12, 2013
• Decision: SESE
• Submission Type: Traditional
• Regulation: 21 CFR 862.1155
• Device Class: Class 2

Intended Use

The Co-Innovation One Step hCG Test Strip is an in vitro diagnostic visual qualitative immunochromatographic assay designed for the rapid determination of human chorionic gonadotropin (hCG) in urine to aid in the early detection of pregnancy. It is intended for over-the-counter (OTC) use. The Co-Innovation One Step hCG Test Cassette is an in vitro diagnostic visual qualitative immunochromatographic assay designed for the rapid determination of human chorionic gonadotropin (hCG) in urine to aid in the early detection of pregnancy. It is intended for over-the-counter (OTC) use. The Co-Innovation One Step hCG Test Midstream is an in vitro diagnostic visual qualitative immunochromatographic assay designed for the rapid determination of human chorionic gonadotropin (hCG) in urine to aid in the early detection of pregnancy. It is intended for over-the-counter (OTC) use.

Device Story

Rapid sandwich immunoassay for qualitative hCG detection in urine; aids early pregnancy diagnosis. Available in strip, cassette, and midstream formats. Operates via colloidal gold-labeled mouse monoclonal anti-beta-hCG antibodies on a conjugate pad and immobilized mouse monoclonal anti-alpha-hCG antibodies on a nitrocellulose membrane. Urine sample application (dip, dropper, or midstream) initiates capillary flow; hCG binds to gold-conjugate, forming complexes captured at the test line (T) by anti-alpha-hCG antibodies, creating a visible red band. Control line (C) confirms proper flow. Used in home settings by lay users; results read visually after 5 minutes. Clinical decision-making relies on presence/absence of test line. Benefits include rapid, convenient, at-home pregnancy screening.

Clinical Evidence

Bench testing only. Precision/reproducibility evaluated across 3 lots and 3 sites using spiked urine samples (0-100 mIU/mL). Lay-user study (n=353) demonstrated 100% agreement with professional testing across all formats (strip, cassette, midstream). Analytical specificity confirmed no interference from common endogenous/exogenous compounds, hCG beta-core fragments, or cross-reactivity with LH, FSH, or TSH. No high-dose hook effect observed up to 1,000,000 mIU/mL.

Technological Characteristics

Lateral flow immunoassay; double antibody sandwich principle. Materials: nitrocellulose membrane, mouse monoclonal anti-beta-hCG antibody-colloidal gold conjugate, mouse monoclonal anti-alpha-hCG antibody, goat anti-mouse IgG. Formats: strip, cassette (plastic housing), midstream (plastic housing). Qualitative visual readout. Traceable to WHO 3rd International Standard. No electronic components or software.

Indications for Use

Indicated for the rapid, qualitative detection of human chorionic gonadotropin (hCG) in human urine to aid in early pregnancy detection. Intended for over-the-counter (OTC) use by individuals aged 18-45.

Regulatory Classification

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. 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.

Predicate Devices

• One Step HCG Urine Pregnancy Test (K071930)

Submission Summary (Full Text)

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510(k) SUBSTANTIAL EQUIVALENCE DETERMINATION
DECISION SUMMARY
ASSAY ONLY TEMPLATE

A. 510(k) Number:
k132085

B. Purpose for Submission:
New device

C. Measurand:
Human chorionic gonadotropin

D. Type of Test:
Lateral flow immunoassay

E. Applicant:
Co-Innovation Biotech Company Ltd.

F. Proprietary and Established Names:
Co-Innovation One Step hCG Test Strip
Co-Innovation One Step hCG Test Cassette
Co-Innovation One Step hCG Test Midstream

G. Regulatory Information:
1. Regulation section:
Human chorionic gonadotropin 21 CFR 862.1155
2. Classification:
Class II
3. Product code:
LCX

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4. Panel:
Clinical Chemistry (75)

H. Intended Use:

1. Intended use(s):
See indications for use.

2. Indication(s) for use:
The Co-Innovation One Step hCG Test Strip is an in vitro diagnostic visual qualitative immunochromatographic assay designed for the rapid determination of human chorionic gonadotropin (hCG) in urine to aid in the early detection of pregnancy. It is intended for over-the-counter (OTC) use.

The Co-Innovation One Step hCG Test Cassette is an in vitro diagnostic visual qualitative immunochromatographic assay designed for the rapid determination of human chorionic gonadotropin (hCG) in urine to aid in the early detection of pregnancy. It is intended for over-the-counter (OTC) use.

The Co-Innovation One Step hCG Test Midstream is an in vitro diagnostic visual qualitative immunochromatographic assay designed for the rapid determination of human chorionic gonadotropin (hCG) in urine to aid in the early detection of pregnancy. It is intended for over-the-counter (OTC) use.

3. Special conditions for use statement(s):
For over-the-counter use

4. Special instrument requirements:
Not applicable

I. 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-α-hCG antibody (on the Test Line) and goat anti mouse IgG polyclonal antibody (on the Control Line).

J. Substantial Equivalence Information:

1. Predicate device name(s):

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Blue Cross Biomedical One-Step hCG Urine Pregnancy Test

2. Predicate 510(k) number(s):

k071930

3. Comparison with predicate:

| Feature | New Device | Predicate (k071930) |
| --- | --- | --- |
| Intended Use | Qualitative detection of human chorionic gonadotropin (“HCG”) in urine | Same |
| Indications/Intended Users | Over the Counter (OTC) | Over the Counter (OTC) Use and Prescription Use |
| Specimen | Urine | Same |
| Clinical cut-off | 25mIU/mL | Same |
| Read time | 5 minutes | Same |
| Test Principle | Colloidal Gold Immunoassay | Same |
| Traceability | WHO 3rdIS | Same |
| Format | Strip, cassette, midstream | Same |
| Reagents | Monoclonal anti-β hCG antibody colloidal gold conjugate on the pre-dried pad. Monoclonal anti-α hCG antibodies (on the test region) Goat anti mouse IgG (on the control region). | Same, but specific antibodies differ. |

K. Standard/Guidance Document Referenced (if applicable):

Guidance for Over-the Counter (OTC) Human Chorionic Gonadotropin (hCG) 510(k)s

L. Test Principle:

The assay of each device uses a double antibody sandwich method. Each test device contains mouse monoclonal anti-β-hCG antibody colloidal gold conjugate pre-dried on a pad. Mouse monoclonal anti-α-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. During the test procedures, hCG in the urine specimen reacts with the dye conjugate (mouse anti-β-hCG antibody-

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colloidal gold conjugate specific to the beta subunit of hCG) and forms a complex. Because of capillary and chromatographic effects of the nitrocellulose membrane, the complex migrates along the membrane to the $\alpha$ -hCG antibody line (T), and remains 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.

# M. Performance Characteristics (if/when applicable):

# 1. Analytical performance:

# a. Precision/Reproducibility:

Precision studies were performed using urine samples spiked with a standard containing natural intact hCG, purified from urine, and calibrated versus the WHO 3rd standard. Concentrations included $0\mathrm{mIU / mL}$ , $12.5\mathrm{mIU / mL}$ , $18.75\mathrm{mIU / mL}$ , $25\mathrm{mIU / mL}$ , $50\mathrm{mIU / mL}$ , and $100\mathrm{mIU / mL}$ hCG. Samples were masked and randomized prior to testing. The study was conducted over 10 days by multiple operators at 3 hospital sites. Three lots of each of the three formats were tested. The midstream format was tested with both the dip and (simulated) midstream method in this evaluation. Results are shown below:

Strip format test results:

| hCG concentration | Lot 1 | | Lot 2 | | Lot 3 | |
| --- | --- | --- | --- | --- | --- | --- |
| | Positive | Negative | Positive | Negative | Positive | Negative |
| 0 mIU/mL | 0 | 30 | 0 | 30 | 0 | 30 |
| 12.5 | 0 | 30 | 0 | 30 | 0 | 30 |
| 18.75 | 0 | 30 | 0 | 30 | 0 | 30 |
| 25 | 30 | 0 | 30 | 0 | 30 | 0 |
| 50 | 30 | 0 | 30 | 0 | 30 | 0 |
| 100 | 30 | 0 | 30 | 0 | 30 | 0 |

Cassette format test results:

| hCG concentration | Lot 1 | | Lot 2 | | Lot 3 | |
| --- | --- | --- | --- | --- | --- | --- |
| | Positive | Negative | Positive | Negative | Positive | Negative |
| 0 mIU/mL | 0 | 30 | 0 | 30 | 0 | 30 |
| 12.5 | 0 | 30 | 0 | 30 | 0 | 30 |
| 18.75 | 0 | 30 | 0 | 30 | 0 | 30 |
| 25 | 30 | 0 | 30 | 0 | 30 | 0 |
| 50 | 30 | 0 | 30 | 0 | 30 | 0 |
| 100 | 30 | 0 | 30 | 0 | 30 | 0 |

Midstream (dip) format test results:

| hCG concentration | Lot 1 | | Lot 2 | | Lot 3 | |
| --- | --- | --- | --- | --- | --- | --- |
| | Positive | Negative | Positive | Negative | Positive | Negative |
| 0 mIU/mL | 0 | 30 | 0 | 30 | 0 | 30 |

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| 12.5 | 0 | 30 | 0 | 30 | 0 | 30 |
| --- | --- | --- | --- | --- | --- | --- |
| 18.75 | 0 | 30 | 0 | 30 | 0 | 30 |
| 25 | 30 | 0 | 30 | 0 | 30 | 0 |
| 50 | 30 | 0 | 30 | 0 | 30 | 0 |
| 100 | 30 | 0 | 30 | 0 | 30 | 0 |

Midstream (simulated midstream) test results

| hCG concentration | Lot 1 | | Lot 2 | | Lot 3 | |
| --- | --- | --- | --- | --- | --- | --- |
| | Positive | Negative | Positive | Negative | Positive | Negative |
| 0 mIU/mL | 0 | 30 | 0 | 30 | 0 | 30 |
| 12.5 | 0 | 30 | 0 | 30 | 0 | 30 |
| 18.75 | 0 | 30 | 0 | 30 | 0 | 30 |
| 25 | 30 | 0 | 30 | 0 | 30 | 0 |
| 50 | 30 | 0 | 30 | 0 | 30 | 0 |
| 100 | 30 | 0 | 30 | 0 | 30 | 0 |

Lay user precision:

Spiked urine samples (prepared in the same way as those described above) were also tested by lay persons. All aliquots were masked and randomized prior to testing. The lay user participants included various levels of educational backgrounds and ages (18 to 45 years). Each subject conducted one test, using the English package insert as guide. Results are shown below:

| Format | Strip | | Cassette | | Midstream | |
| --- | --- | --- | --- | --- | --- | --- |
| HCG Concentration | Positive | Negative | Positive | Negative | Positive | Negative |
| 18.75mIU/ml | 0 | 100 | 1 | 99 | 0 | 100 |
| 31.25mIU/ml | 100 | 0 | 100 | 0 | 99 | 1 |
| Total | 100 | 100 | 101 | 99 | 99 | 101 |

b. Linearity/assay reportable range:

Not applicable. This is a qualitative assay.

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

The 510(k) describes traceability of the assay to the WHO 3rd reference material. Stability testing information (protocols and acceptance criteria) to support the claimed shelf life (2 years) was reviewed and deemed acceptable.

d. Detection limit:

See Precision section above (M.1.a.).

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# e. Analytical specificity:

The assay was tested for interference from endogenous compounds, potentially interfering clinical conditions, and exogenous compounds in the presence of $0\mathrm{mIU / mL}$ hCG and $25\mathrm{mIU / mL}$ hCG. Three lots were tested. No interference was observed from the compounds at the concentrations listed below.

| Substances tested/ concentration | |
| --- | --- |
| Acetaminophen | 20mg/dL |
| Aspirin | 20mg/dL |
| Ascorbic acid | 20mg/dL |
| Atropine | 20mg/dL |
| Caffeine | 20mg/dL |
| Glucose | 2000mg/dL |
| Hemoglobin | 500mg/dL |
| Tetracycline | 20mg/dL |
| Ampicillin | 20mg/dL |
| Albumin | 2000mg/dL |
| Bilirubin | 2mg/dL |
| Acetaminophen | 20mg/dL |
| Aspirin | 20mg/dL |
| Ascorbic acid | 20mg/dL |
| Atropine | 20mg/dL |
| Caffeine | 20mg/dL |
| Glucose | 2000mg/dL |
| Hemoglobin | 500mg/dL |
| Tetracycline | 20mg/dL |
| Ampicillin | 20mg/dL |
| Albumin | 2000mg/dL |
| Bilirubin | 2mg/dL |
| Leukocyte | >500/ul |
| Erythrocytes | >250/uL |
| Uric acid | 0.58 mMol/L |
| Ketone | >80 mg/dL |

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To evaluate any effects of the hCG $\beta$ -core fragment, urine from non-pregnant females containing 0 and $25\mathrm{mIU / ml}$ hCG were spiked with the hCG $\beta$ -core fragment (traceable to WHO reference reagent 99/708) to concentrations of 125,000, 250,000, 500,000 and 1,000,000 pmol/mL. Three batches of each format were tested. No effects were seen for the concentrations tested. Results are shown below:

| hCGβ-core Fragment concentration (pMol/L) | hCG concentration | Batch 1 | | Batch 2 | | Batch 3 | |
| --- | --- | --- | --- | --- | --- | --- | --- |
| | | Positive | Negative | Positive | Negative | Positive | Negative |
| 125000 | 0mIU/ml | 0 | 1 | 0 | 1 | 0 | 1 |
| 250000 | 0mIU/ml | 0 | 1 | 0 | 1 | 0 | 1 |
| 500000 | 0mIU/ml | 0 | 1 | 0 | 1 | 0 | 1 |
| 1000000 | 0mIU/ml | 0 | 1 | 0 | 1 | 0 | 1 |
| 125000 | 25mIU/ml | 1 | 0 | 1 | 0 | 1 | 0 |
| 250000 | 25mIU/ml | 1 | 0 | 1 | 0 | 1 | 0 |
| 500000 | 25mIU/ml | 1 | 0 | 1 | 0 | 1 | 0 |
| 1000000 | 25mIU/ml | 1 | 0 | 1 | 0 | 1 | 0 |

To evaluate for high dose hook effect, urine specimens spiked with hCG at concentrations shown below were tested across three lots. No hook effect was observed. Results are shown below:

| hCG | Positive | Negative |
| --- | --- | --- |
| 62,500mIU/ml | 9 | 0 |
| 125,000mIU/ml | 9 | 0 |
| 250,000mIU/ml | 9 | 0 |
| 500,000mIU/ml | 9 | 0 |
| 1,000,000mIU/ml | 9 | 0 |

Evaluations of the effect of the pH and specific gravity of urine specimens were also performed. Results of samples containing 0 and $25\mathrm{mIU / mL}$ hCG were unaffected across the pH range of 3-9 and specific gravity of 1.01-1.04.

Evaluations were performed to determine cross-reactivity with LH, FSH, TSH. No cross-reactivity was observed over 3 lots. Results are shown below for one representative lot.

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| hormone concentration | hCG concentration | Positive | Negative |
| --- | --- | --- | --- |
| 500mIU/ml LH | 0mIU/ml HCG | 0 | 30 |
| 1000mIU/ml FSH | 0mIU/ml HCG | 0 | 30 |
| 1000uIU/ml TSH | 0mIU/ml HCG | 0 | 30 |
| 500mIU/ml LH | 25mIU/ml HCG | 30 | 0 |
| 1000mIU/ml FSH | 25mIU/ml HCG | 30 | 0 |
| 1000uIU/ml TSH | 25mIU/ml HCG | 30 | 0 |

f. Assay cut-off:
See Precision section above (M.1.a.). The assay cut-off of all devices is 25 mIU/mL.

2. Comparison studies:

a. Method comparison with predicate device:

A method comparison study was conducted where results of professional users with the candidate test were compared to results of professional users with the predicate device. Samples from 353 women were tested.

| Cassette test | | Predicate Device (tested by professional) | |
| --- | --- | --- | --- |
| | | Positive result | Negative result |
| Candidate One step hCG test cassette method (tested by professional) | Positive result | 39 | 0 |
| | Negative result | 0 | 74 |
| Strip test | | Predicate Device (tested by professional) | |
| --- | --- | --- | --- |
| | | Positive result | Negative result |
| Candidate One step hCG test strip method (tested by professional) | Positive result | 38 | 0 |
| | Negative result | 0 | 42 |

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| Midstream test - dip method | | Predicate Device (tested by professional) | |
| --- | --- | --- | --- |
| | | Positive result | Negative result |
| Candidate One step hCG test midstream dip method (tested by professional) | Positive result | 37 | 0 |
| | Negative result | 0 | 43 |
| Midstream test - simulated midstream | | Predicate Device (tested by professional) | |
| --- | --- | --- | --- |
| | | Positive result | Negative result |
| Candidate One step hCG test midstream simulated method (tested by professional) | Positive result | 41 | 0 |
| | Negative result | 0 | 39 |

b. Matrix comparison:

Not applicable.

3. Clinical studies:

a. Clinical Sensitivity:

Not applicable.

b. Clinical specificity:

Not applicable.

c. Other clinical supportive data (when a. and b. are not applicable):

A lay-user study was conducted with 353 layer users who tested their own urine using instructions contained in the English package insert. This

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included 80 layer users using the test strip, 113 using the test cassette, and 160 users using the midstream test (80 using either the dip method or the midstream method). Approximately half of the women were less than 5 weeks pregnant (based on last menstrual period). Samples were randomly collected at various times throughout the day. Subject ages ranged from 18 to 45 years. Subjects responded to questionnaires after finishing the test and collected samples for tests by laboratory professionals using the candidate device, and indicated that they found the test easy to perform. Samples were masked and randomized prior to professional testing. Results are shown below:

| Cassette test | | Candidate Device (tested by professional) | |
| --- | --- | --- | --- |
| | | Positive result | Negative result |
| Candidate One step hCG test cassette method (tested by lay user) | Positive result | 39 | 0 |
| | Negative result | 0 | 74 |
| Strip test | | Candidate Device (tested by professional) | |
| --- | --- | --- | --- |
| | | Positive result | Negative result |
| Candidate One step hCG test strip method (tested by lay user) | Positive result | 38 | 0 |
| | Negative result | 0 | 42 |
| Midstream test - dip method | | Candidate Device (tested by professional) | |
| --- | --- | --- | --- |
| | | Positive result | Negative result |
| Candidate One step hCG test midstream dip method (tested by lay user) | Positive result | 37 | 0 |
| | Negative result | 0 | 43 |

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| Midstream test – midstream method | | Candidate Device (tested by professional) | |
| --- | --- | --- | --- |
| | | Positive result | Negative result |
| Candidate One step hCG test midstream, midstream method (tested by lay user) | Positive result | 41 | 0 |
| | Negative result | 0 | 39 |

4. Clinical cut-off:
Not applicable.

5. Expected values/Reference range:
Not applicable.

N. Proposed Labeling:
The labeling is sufficient and it satisfies the requirements of 21 CFR Part 809.10.

O. Conclusion:
The submitted information in this premarket notification is complete and supports a substantial equivalence decision.