Simple hCG - Pregnancy test Operon - Free user manual and instructions

Product type	Single-use urine pregnancy test
Detection method	Immunochromatographic detection of hCG hormone
Sensitivity	25 mIU/mL
Reliability	99% from the expected day of menstruation
Result	Visible in 3 to 5 minutes
Reading	Control line and test line
Dimensions	Approximately 8 cm x 2.5 cm
Weight	Approximately 5 g
Power supply	None (non-electronic test)
Storage	Room temperature (2-30°C), away from moisture
Expiry date	Indicated on packaging
Number of tests per box	1 test
Recommended use	From the first day of missed period
Usage precautions	Do not use after expiry date; read instructions before use
Maintenance and cleaning	No maintenance required; discard after use
Safety	External use only; do not swallow
Spare parts and repairability	Not repairable; single-use product
General information	Brand Operon, model Simple hCG

Frequently Asked Questions - Simple hCG Operon

How to use the Operon Simple hCG pregnancy test?

Dip the strip in a urine sample for 5 seconds, then place it flat. Read the result after 3 to 5 minutes. Do not interpret after 10 minutes.

What time of day should the test be done?

It is recommended to use the first morning urine as it contains the highest concentration of the hCG hormone.

What is the reliability of the Operon Simple hCG test?

The test shows 99% reliability from the expected day of menstruation. If in doubt, repeat the test a few days later.

How to read the test result?

Two colored lines (control and test) indicate a positive result. A single control line indicates a negative result. No line or only a test line means an invalid test.

What to do if the result is positive?

A positive result indicates probable pregnancy. Consult a doctor for confirmation and follow-up.

What to do if the result is negative but menstruation does not occur?

Wait a few days and repeat the test. If the delay persists, consult a healthcare professional.

Can the test be used before a missed period?

It is less reliable before the delay. For optimal accuracy, wait until the expected day of menstruation or after.

How to store the test?

Store the test in its original packaging at room temperature (2-30°C), away from moisture and direct light.

Can the test give a false positive?

Some medications or rare medical conditions can cause a false positive. If in doubt, perform a confirmation test.

Where can I find the complete instructions?

The complete instructions are available for free download in PDF format on the website notice-facile.com.

USER MANUAL Simple hCG Operon

One-step immunochromatographic test for the qualitative detection of human chorionic gonadotropin (hCG) hormone in urine

INTENDED USE

The Simple or Stick hCG urine chromatographic immunoassay is a procedure for in vitro qualitative detection of human chorionic gonadotropin (hCG) hormone in urine.

This test has been developed to offer quick, efficient help in diagnosing pregnancy; it can be performed on the first missed day of a woman's period and provides a result in only 1-3 minutes.

Human Chorionic Gonadotropin (hCG) is a hormone produced by the placenta in pregnant women. It can be detected in urine 7 to 10 days after conception, which makes it the ideal pregnancy indicator.

The test is based on the immunological capture of microparticles that are coloured as they pass through a membrane on which the monoclonal antibody for complete hCG has been immobilized.

PRINCIPLE

The Simple hCG / Stick hCG urine test uses a combination of a monoclonal antibody for hCG conjugated to red latex particles and another monoclonal hCG-specific antibody in the membrane. Blue microspheres are also used as the test control.

In this test, the urine sample is placed directly on the strip; no previous treatment of the urine is necessary.

When the sample flows through the test membrane, the coloured particles migrate. In the case of a positive result, the specific antibodies present in the membrane capture the coloured particles.

Different colour lines will be visible, depending on the hCG concentration in the sample. These lines are used to interpret the result, just after three minutes room-temperature incubation.

KIT CONTENTS

- Reaction devices (cassette or strip)

The number of reaction devices in the kit is indicated on the outer kit label after the symbol

- Ungraduated disposable plastic pipettes (only in the Simple format)

MATERIAL NOT INCLUDED IN THE KIT

• Test tubes

. Pipettes that can be graduated

• Pipette tips

PRECAUTIONS

1. Patient samples may contain infectious agents and should be treated and discarded as potentially dangerous biological materials.

2. Do not store or prepare food, eat, drink or smoke in the area where the reagents and samples are handled.

3. Do not exchange components from kits with different lot numbers.

4. Use all the reagents only in vitro.

5. Before using the reagents, let all kit components and samples reach room temperature, because cold reagents and/or samples can decrease test functionality. About 20-30 minutes are usually sufficient for reaching room temperature.

6. Do not use kit components after the expiration dates.

7. If the package is broken, the product can still be used as long as none of the components have been damaged.

8. It is important to add the correct sample amount. If less than the amount of sample indicated is used, the chromatography may not occur because the sample might not reach the reaction area; if there is too much, the reagent may be diluted and produce a weak line.

9. The used product should be discarded as indicated by current legislation.

10. Do not use the test if a coloured line appears in the result area before you start to use it.

11. In the case of the Stick product packaged in a tube, it is important to close the tube again immediately after taking the reactive strip out; otherwise, high room humidity might damage the rest of the strips inside the tube.

12. Do not discard the external box of the kit until its content has been totally used. The external box contains essential information regarding the EC marking and lotification.

LIMITATIONS

1. This test is to be used only for detecting hCG in urine.

2. This is a qualitative test; no quantitative interpretation of the result should be made with respect to the intensity of the positive line.

3. The results of the test should be used together with other information available from the patient's clinical evaluation and other diagnostic procedures.

4. It is important to add the correct sample amount. If less than indicated is used, the chromatography may not occur because the sample might not reach the reaction area; if there is too much, the reagent may be diluted and produce a weak line.

5. It is important to control reaction time. If reaction time is less than that indicated, samples with an analyte amount higher than the sensitivity limit can be seen clearly, but those that are at the limit will not appear. If reaction time is greater than that indicated, test sensitivity will be altered and erroneous interpretations may result.

6. The analysis of some samples can produce lines of undefined colour, caused by negative samples in the majority of the cases. The test should be repeated if these lines of undefined colour appear. If the same result is obtained again, performing the analysis with another analytic method is recommended.

STORAGE

The kit can be kept at temperatures between 2^irc C and 30^irc C. The expiration date is stamped on the package.

SAMPLES

Urine samples can be collected in plastic or glass containers at any time of day. However, it is best to use the first morning urine because it usually contains the highest concentration of hCG hormone.

Samples can be kept in the refrigerator for 1-2 days. To conserve them for a longer period, they should be frozen at -20°C. Before frozen samples are used in the test, they should be unfrozen, letting them reach room temperature.

PROCEDURE

SIMPLE HCG URINE PROCEDURE

1. Take the test out of the protective pouch just before using it.
2. Add 4 drops or 125 µ l of urine slowly to the round window marked with an arrow.
3. Check the result after 3 minutes.

Depending on the hCG concentration, a positive result may be seen in under a minute.

STICK HCG URINE PROCEDURE

A. By immersion:

1. Place 0.5 ml of urine in a test tube or vial.
2. Place the Stick hCG inside, in a vertical position, with the white tip in the urine. The urine should never reach the area with the arrows.

B. By sample absorption:

1. Introduce the reactive strip in the urine up to the arrows for 5-10 seconds.
2. Place the strip on a clean, dry, flat surface.

Check the result after 3 minutes for both procedures. Most positive results appear after 1 minute. Note: When the strip is submerged in the urine sample, it should never reach the area with the arrows.

READING RESULTS

NEGATIVE: There is only a BLUE line in the results area (in the SIMPLE test, aligned with the letter 'C' (Control) marked on the device frame). This line should always appear.

POSITIVE: Two bands or lines appear in the results area: a RED/PINK line (positive) and aBLUE line (control). The highest line (the blue control one) will always appear as a strong line. The intensity of the colour of the lower, red line (the result line) depends on the concentration of hCG in the sample. This intensity can vary from very strong (when the hCG concentration is high) to weak (when the concentration is close to the sensitivity limit of the test).

INVALID: No line appears in the results area. Other possible indicators of invalid results are two bands of the same colour (two red or two blue lines) or a single red line. Invalid results can be caused by deterioration of the reagents or incorrect handling. In the case of an invalid result, the test should be repeated with new strips.

Any line that may appear after 5 minutes due to the nature of the test has no diagnostic value.

Very high hCG concentrations may be found in urine from patients having chorionic epithelioma or hydatidiform mole.

The final diagnosis should not be based on only the result from one test. The diagnosis should be established by the correlation of test results with other appropriate data and with clinical symptoms. The interpretation of the results is as follows:

Invalid

Negative

Positive

QUALITY CONTROL

The test is invalid if no line appears or only a red line appears (without the blue one), because the test was not performed properly or the reagents have deteriorated.

WARNING: Including a control with an established result is recommended to ensure that the data obtained are correct.

SENSITIVITY

Based on numerous tests with various observers under different conditions and with different lots, a standard sensitivity of 25 mIU/ml has been established for hCG. However, amounts of up to 6.25 mIU/ml have been detected in the best of cases.

DIAGNOSTIC SENSITIVITY AND SPECIFICITY

TEST 1

A comparative evaluation carried out on our product and the one taken as the reference product, the ELISA microliter hCG Mab IEMA (Equipar Diagnostici, Italy), is presented.
The evaluation was performed at Operon S.A. on 208 urine samples provided by a hospital in Zaragoza, Spain.
The results were as follows:

208 urine samples		ELISA microliter hCG Mab IEMA
		+	-
Operon Stick hCG urine	+ 37	1
	- 0 1	70

Concordance percent: 100% [(37+170)/208] = 99.5%

Estimated sensitivity concordance: 100% [37/(37+0)] = >99%

Estimated specificity concordance: 100%[170(170+1)] = 99.4%.

INTRA-ASSAY PRECISION

Five replicas of each concentration of the sensitivity curve were tested with one lot of hCG urine; similar results were obtained, differing only in a 1:2 dilution in the most extreme cases (sensitivity 25-12.5 mIU/ml).

REPRODUCIBILITY

INTER-DAY PRECISION

Using the same product lot, 4 replicas of the sensitivity curve were performed over 4 consecutive days. The results were completely reproducible (sensitivity of 25 mIU/ml in all cases).

INTER-OPERATOR PRECISION

Four people, with no previous training, carried out the test using a serial dilution of the internal Operon standard. Only a difference of a 1:2 dilution was found, which is acceptable and tolerable for the test performed.

INTER-LOT PRECISION

A sensitivity curve was performed in duplicate with 8 product lots. The analyses were preformed by the same person on the same day. There was 100% reproducibility (sensitivity of 12.5 mIU/ml.)

Observed differences are acceptable and tolerable as it is a qualitative immunochromatographic technique, with an inherent variability.

HOOK EFFECT

Various samples that contained different amounts of hCG (250 - 500 - 750 - 1000 - 1500 - 2000 IU/ml) were tested with 3 different lots of Stick hCG urine. It should be pointed out that 2,000,000 mIU hCG/ml is an amount that is several times greater than the maximum hCG concentration found in pregnant women.

The values were clearly positive in all tests, even at the highest concentration analyzed. It can thus be concluded that concentrations of up to 2 × 10^-6 mIU/ml can be detected with Stick hCG urine.

INTERFERING SUBSTANCES

The substances indicated in the table, at the concentration specified, did not interfere with the result.

Phenylpropanolamine (200 mg/l) Caffeine (200 mg/l)

Hemoglobin (10 mg/ml) Gentisic Acid (200 mg/l)

Acetaminophen (200 mg/l) Glucose (20 mg/ml)

Acetylsalicylic Acid (200 mg/l) Bilirubin (0.02 mg/ml)

Ampicillin (200 mg/l) Protein (20 mg/ml)

Ascorbic Acid (200 mg/l) Estriol (0.02 mg/ml)

Atropine (200 mg/l) Pregnane diol (0.02 mg/ml)

CROSS REACTIONS

No cross reactions were observed with the following hormones at the concentrations normally found in physiological conditions (even in extreme cases).

hormone	Conc. mIU/ml	Lot 00211
		1'	3'	5'
hLH	1000	-	-	-
hLH	750	-	-	-
hLH	500	-	-	-
hLH	375	-	-	-
hLH	250	-	-	-
hLH	0 -	-	-
hFSH 2000	-	-	-
hTSH	11	-	-	-

FR

SIMPLE hCG / STICK hCG urine

• S. Brody S and G. Carlstrom, Immunoassay of Human Chorionic Gonadotropin in Normal and Pathologic Pregnancy, J. Clin. Endocrinology and Metabolism, 22, 564 (1962)
• Robert O. Hussa: The Clinical Marker hCG. Praeger Publishers, New York 1987.
• Siegfried Schwartz, Peter Berger and Georg Wick: The Antigenic Surface of Human Chorionic Gonadotropin as Mapped by Murine Monoclonal Antibodies, Endocrinology 118(1), 189-197 (1986)
• Laurence A. Cole, David B. Seifer, Andrew Kardana and Glenn D. Braunstein, Selecting human chorionic gonadotropin immunoassays: Consideration of cross-reacting molecules in first-trimester pregnancy serum and urine, Am. J. Obstet. Gynecol., 168, 1580-1586 (1993)
• Laurence A. Cole, Immunoassay of human Chorionic Gonadotropin, its free subunits and metabolites, Clin. Chem., 43 (12), 2233-2243 (1997)
• Ulf-Hakan Stenman, Leila Unkila-Kallio, Juha Korhonen and Henrik Alfthan, Immunoprocedures for detecting human chorionic gonadotropin: clinical aspects and doping control, Clin. Chem., 43(7), 1293-1298 (1997)
• S.A. Butler, S.A. Khanlian and L.A. Cole, Detection of Early Pregnancy Forms of Human Chorionic Gonadodropin by Home Pregnancy Test Devices, Clin. Chem., 47(12), 2131-2136 (2001)