IFU of HCG Dual Threshold Test--MIDSTREAM


INTENDED USE

David One Step Pregnancy Urine Test is an in-Vitro diagnostic test for the semi-quantitative detection of human chorionic gonadotropin (HCG) in women's urine to prediction of the number of week in early pregnancy.

PRINCIPLE

The product adopts the principle of immunochromatographic double antibody sandwich method and colloidal gold labeling technology. It is made of nitrocellulose membrane respectively solid phased with mouse anti-α-HCG monoclonal antibody (test line, T line), mouse anti-HCG antibody (reference line, R line) and goat anti-mouse IgG antibody (control line, C line), colloidal gold-labeled mouse anti-β-HCG monoclonal antibody and other components.

At the time of detection, human chorionic gonadotropin (HCG) in urine sample combine with gold-labeled mouse anti-β-HCG monoclonal antibody   pre-adsorbed on the kit to form “HCG- (colloidal gold-labeled mouse anti- β-HCG monoclonal antibody)” complex, which continues to move forward by the capillary action and then in turn combine with the mouse anti-α-monoclonal HCG antibody on the test zone (T-line) and mouse anti-HCG antibody on the reference zone (R line) to form a double antibody sandwich complex “(mouse anti-α-HCG monoclonal antibody-HCG- (colloidal gold-labeled mouse anti-β-HCG monoclonal antibody), (mouse anti-HCG antibody)-HCG- (colloidal gold-labeled mouse anti-β-HCG monoclonal antibody)” . A red line appears respectively in the test zone(T line) and reference zone (R line).   

The sensitivity of the test line (T line) is high while and the sensitivity of the reference line (R line) is low. If a red line appears in the test zone (T line), the result is positive, indicating that HCG is detected in the urine sample. The reminder of pregnant time length is estimated according to the HCG content in urine. The contents of HCG in urine samples of pregnant women at different pregnant time points are different. At the 1st to the 3rd week of pregnancy, only one red line appears at the test zone (T). As the number of weeks of pregnancy increases (not more than 8 weeks), the amount of HCG in the urine increases continuously. When the pregnancy is more than 3 weeks, the test line (T line) and the reference line (R line) simultaneously develop color.

PRECAUTIONS

1. For in vitro diagnostic use (not for internal use).

2. Do not use this test past the indicated expiry date.

3. The test device must remain in the sealed pouch until use.

4. Do not use if the pouch is torn or not well sealed.

5.Do not touch the membrane located within the window and Absorbent Tip.

6.The test should not be reused.

MATERIALS

Reagent and Material Supplied

Each test kit individually sealed in a foil pouch. Each pouch contains: 1. One HCG Test Device,   2. a desiccant.

NOTE: The desiccant is only for storage and should not be used during the procedure.

Material Required but not Supplied

1.Timer      2. A clean container for a urine sample collection (optical)

No other equipment or reagents are needed.

STORAGE AND STABILITY

1.The test kit can be Stored at 4 to 30℃ in the sealed pouch. DO NOT FREEZE. The test is stable through the expiration date printed on the pouch label or label of the outer box.

2.Preferably open the pouch only shortly before the test.

SPECIMEN COLLECTION AND PREPARATION

When to Collect Urine for Test

If testing from the day your period is due, you can test any time of day.If testing early, use the first-morning urine of the day.

How to Collect Urine

1.Specimens collected at any time of day may be used; however, the first morning urine generally contains the highest concentration of HCG and are recommended for early detection of pregnancy.

2. If you do not want to test directly with the urine stream, use a clean, dry glass or plastic container to collect urine.

3.Fresh urine does not require any special pretreatment.

Specimen Storage

Specimens may be kept at room temperature for 8 hours or stored at 2℃8℃ for up to 3 days.   For prolonged storage, specimens may be frozen and stored below -20℃. Frozen specimens should be thawed and mixed before testing.

TEST PROCEDURE

If stored refrigerated, ensure that the pouched device, and specimen is brought to room temperature about 20 minutes before use.

1.Remove the test device from the sealed pouch and use it as soon as possible.

2.Hold the stick by the thumb grip with the exposed Absorbent Tip pointing downward. Urinate on the Absorbent Tip only until it is thoroughly wet {about 2-3seconds}.

Note: Do not urinate on the Test and Control Windows.

If you prefer, you can urinate into a clean and dry container, then dip only the Absorbent Tip of the midstream test into the urine for at least 10 seconds. Avoid cross-contamination of specimens by using a new specimen collection container for each specimen obtained.

3. Lay the test device down on a flat surface.

4.Start the time and wait for the colored line(s) to appear.   Read the result at 5 minutes. Do not read the results after 15 minutes.

NOTE: Depending on the concentration of hCG in the test specimen, positive results may be observed as soon as 10-30 seconds. However, to confirm negative results, the complete reaction time of 5 minutes is required.   

INTERPRETATION OF RESULTS

Pay attention to the position of the Control Line(C), reference line (R) and Test Line (T).

Negative Result means that no hCG has been detected in your urine. You may not be pregnant, or the level of pregnancy hormone may not be high enough yet to be detected. If you still have reason to believe that you are pregnant (no rules, for example), you can repeat the test a few days later. If the test is still negative, consult your doctor.

Positive Result means that your urine contains hCG and you are probably pregnant. The intensity of the color of the bands may vary depending on the different levels of hCG concentration at various stages of pregnancy. You should consult your doctor. He can confirm your pregnancy after analyzing the laboratory results.


Item

Test Result

Conceiving Time

Clinical gestational age

Invalid Result

At   5 minutes, if no red line appears in the control line (C) area

/

/

Negative Result

Only the control line develops color.

Both the test line and reference line don’t   develop color.

-

-

Positive Result

Both the control line and test line develop   color.The reference line doesn’t develop color.

about 1-3 weeks

3-5 weeks

Positive Result

The control line, the test line and reference line   develop color.

more than 3 weeks

more than 5 weeks



Note :The number of weeks indicated by this kit is calculated after fertilization, so you can estimate your medical gestational age based on the indicated gestational age. Clinical gestational age refers to the period from the first day of the last menstrual period (not counting from the day of having coitus) to the end of childbirth. Therefore, you can calculate your due date by adding 2 weeks to the gestational age determined by the kit(base on your own ovulation period). If your menstrual cycle persists 28 days, you are expected to be pregnant about 2 weeks before the start of the next menstrual period (ie, egg fertilization).

QUALITY CONTROL

A procedural control is included in the test. A colored line appearing in the Control Line region (C) is considered an internal procedural control. It confirms sufficient specimen volume and correct procedural technique. A clear background is an internal negative procedural control. If a background color appears in the result window and interferes with the ability to read the test result, the result may be invalid.

LIMITATIONS

1.The contents of this kit are for use in the qualitative detection of hCG in urine only, neither the quantitative value nor the rate of increase in hCG can be determined by this test.

2.This test may produce false negative results. In case pregnancy is suspected and the test continues to produce negative results, see a physician for further diagnosis.

--A false negative result (negative when pregnancy exists) may occur if the urine is too dilute or with a very early stage pregnancy. If pregnancy is still suspected, retest using a fresh urine specimen at least 48 hours after initial testing.

3.This test may produce false positive results. The presence of hCG in urine should not be used to diagnose pregnancy unless these conditions have been ruled out.

--Fertility drugs containing   hCG can give misleading results (these fertility drugs are usually given by injection and testing too soon after administration may give a false ‘Pregnant’ result).

--hCG may remain detectable for a few days to several weeks after delivery, spontaneous abortion or termination.

-- A number of medical conditions other pregnancy, including Ectopic pregnancies, ovarian cysts, menopause can cause elevated levels of HCG, which can result in a false or irregular result.

--A number of conditions other than pregnancy including trophoblastic disease and certain non-trophoblastic neoplasms, including testicular tumours, prostate cancer, breast cancer and lung cancer, cause elevated levels of hCG.   

--A conception in the uterus that is lost in the early days of implantation, such as a
chemical pregnancy, might initially give a positive test. When the pregnancy becomes nonviable, usually because of the failure of implantation, the low level of HCG might still be detectable on a blood test for a short while. A repeat pregnancy test will be negative a few days later.

4.This test may produce false Invalid results. Immunologically interfering substances such as those used in antibody therapy treatments may invalidate the test result.

5.Alcohol may interfere the test result. It is not recommended using the test after drinking.

6.Other treatments for fertility (such as clomifen citrat), pain relievers or hormonal contraceptives are not expected to interfere with the result.

7.HCG concentration may be different for different gestational age and different pregnant women, and there is no comparability. Therefore, the gestational age can only be predicted and should be confirmed in combination with own menstrual cycle.

8.If your test result is positive and the predicted gestational age is the same as your expectation (based on ovulation period), please go to the hospital to see a doctor and confirm the diagnosis. The doctor will give you further guidance.

9. When your test result is positive and the predicted gestational age is less than 6 weeks but gradually decrease (or the color of reference line is becoming lighter), which is not expected, you should consult a specialist.

10. If this product is used when your gestational age exceeds 6 weeks, the predicted age may not meet the expected result. This is because the HCG content in the urine is too high, and it is difficult to reliably estimate the time of conception.

11.As with all diagnostic tests, a definitive clinical diagnosis should not be based on the results of a single test, but should only be made by the physician after all clinical and laboratory findings have been evaluated.

EXPECTED VALUES

1.Negative results are expected in healthy non-pregnant women.

2.The amount of hCG will vary greatly with gestational age and between individuals.

PERFORMANCE CHARACTERISTICS

1.Sensitivity: This sensitivity level has been confirmed with hCG standards in urine.At the 5th minute, the cut-off of test line (T line) should not be higher than HCG 10 mIU/mL, the threshold of reference line ( R line) should be HCG 2000 mIU/mL.

2.Specificity: The test was determined from cross reaction studies with known amounts of luteinizing hormone (LH), follicle stimulating hormone (FSH), and thyroid stimulating hormone (TSH). The addition of LH (500 mIU/mL), FSH (1,000 mIU/mL), and TSH (1,000 mIU/mL) to hCG negative and positive specimens showed no cross-reactivity.

3.Interference study :The following potentially interfering substances were added to hCG negative and positive specimens.


Substance
Concentration
Substance
Concentration
Substance
Concentration
Acetaminophen
20 mg/dL
Ephedrine
20mg/dL
Glucose
2g/dL
Atropine
20 mg/dL
Thiophene
20 mg/dL
Albumin
2g/dL
Aspirin
20 mg/dL
Phenylpropanolamine
20 mg/dL
estriol
1mg/dL
Ascorbic Acid
20 mg/dL
Acetylsalicylic Acid
20 mg/dL
Bilirubin
2mg/dL
Ampicillin
20mg/dL
Salicyclic Acid
20 mg/dL
Hemoglobin
1mg/dL
Caffeine
20 mg/dL
Phenothiazine
20 mg/dL
hCG β-core fragment
1,000,000 pmol/L
Gentisic Acid
20 mg/dL
Tetracycline
20mg/dl
Urine PH value
49
Ethanol
1%
urine specific gravity
1.000~1.035



At the levels tested, none of these substances interfered with the expected test results.

4.Accuracy

A multi-center clinical evaluation was conducted comparing the results obtained using the Runbio One Step Pregnancy Test to another commercially available urine membrane hCG test (predicate device). The study included 600 urine specimens. The results demonstrated a 100% overall agreement (for an accuracy of > 99%). Results are summarized below:   



predicate device



Positive
Negative

David Pregnancy Test
Positive
a278
b0

Negative
c0
d322


5.High dose effect

Normal urine that were spiked with hCG concentrations of 62,500, 125,000, 250,000, 500,000, 1,000,000, and 2,000,000 IU/L were used to study the high dose hook effect on Pregnancy Test. It was noticed that both color bands at the test band region and the control region were visible.All samples yield the expected positive results.

6.Standardization

The test has been standardized to the   WHO International Standard 5th WHO Chorionic Gonadotrophin NIBSC code: 07/364.

REFERENCES

1.Johnson SR, Miro F, Barrett S, Ellis JE. Levels of urinary human chorionic gonadotrophin (hCG) following conception and variability of menstrual cycle length in a cohort of women attempting to conceive. Curr Med Res Opin. 2009 Mar; 25(3):741-8

2.Pike J, Ellis J, Johnson S, Shaw R, Parkinson P, Perry P. Accuracy of new Clearblue Digital Pregnancy Test with Conception Indicator. Hum Reprod 2010; 25(suppl 1):i169-170


 RUNBIO BIOTECH CO., LTD.

Rongsheng Technological Zone, Univ.Rd., Shantou, Guangdong, China 515063

Tel:+86-754-82528311   

Website:www.runbio-bio.com   

Emailsales@runbio-bio.com


DOCUMENT NUMBER1312.003/HCG-DM-B

VERSION NUMBERXD 01   

Eff.Date:2018-12-10