Title Diagnostic test kit for the qualitative determination of human Chorionic Gonadotrophin (hCG) in human urine samples.

Objective To determine the presence of human Chorionic Gonadotrophin (hCG) in human urine samples. Introduction A pregnancy test is used to determine whether or not a woman is pregnant. Modern pregnancy tests look for chemical markers associated with pregnancy. These markers are found in urine and blood, and pregna ncy tests require sampling one of these substances. The first of these markers to be discovered, human chorionic gonadotropin (hCG) that is important for all pregnancy test. HCG is a hormone produced during pregnancy. It appears in the blood and urine of pregnant women as early as 10 days after conception. While hCG is a reliable marker of pregnancy, it cannot be detected until after implantation, this results in false negatives if the test is performed during the very early stages of pregnancy. Principle After fertilization, the placenta wil develop immediately and secrete Human Chorionic Gonadotrophin (hCG) is a glycoprotein hormone. At the time of th e first missed menstrual period, hCG concentration in serum and urine are about 100mlU/ml and double in concentration every 1.2 to 2 days. Peak levels of over 100,000mlU/ml hCG are seen late in the first trimester of pregnancy. The early appearance of hCG in urine following conception has made it the marker of choice for the detection of pregnancy. The direct Pregnancy assay is based upon the agglutination reaction between latex particles coated with anti -hCG antibodies and hCG present in the test specimen. The presence of hCG in the specimen results in the formation of an agglutination matrix which is easily visually differentiated from the non-agglutinating negative control. Materials Sample, Control Positive, Control Negative, latex reagent, a gglutination slide, pipette-stirrer

Procedure : 1. First, the latex reagent and specimens are let to room temperature before use. The latex reagent is gently mixed to resuspend latex particles. 2. By using a pipette-stirrer, a drop of urine sample is placed ont o the circle of agglutination slide.

3. The e ve e cy c s ce he he c c e s e. 4. One drop of Pos ve Pre nancy control is placed onto the agglutination slide ne t to the Negative control. 5. After that, the late reagent is shaked and re-suspended. One drop of ate reagent is dropped to each of the test circles of the agglutination slide. 6. All the test circles are stirred by using individual pipette -stirrer and spread mixture over entire area of the test circle. 7. The agglutination slide is rocked for two minutes and the agglutination is observed. Result must be interpreted at two minutes. Extended incubation may result in evaporation and erroneous resluts. Results

No agglutination

Present of agglutination

Specimen Sample Negative pregnancy control Positive pregnancy control

bservation No agglutination No agglutination Presence of agglutination

Positive : agglutination occur Negative : no agglutination

    

  



Discussion : From the test that was performed, the sample and Negative pregnancy control show negative result which mean no agglutination occur when reacting with latex reagent. These show that no hCG hormones presence in both samples and do not react with the latex particles coated with anti-hCG antibodies to produce the agglutination. The result occur because maybe the samples are from healthy men and healthy non-pregnant females who do not have hCG levels detected by this method., For Positive pregnancy control, when the hCG hormones presence in the sample, the latex particles coated with the anti-hCG antibodies will bind together to hCG hormones and produces and become agglutinate. The agglutination between hCG and latex particles with anti-hCG antibodies will form the agglutination matrix and will form visible agglutination on the agglutination slide. The result is positive because the agglutination occur during the reaction. But some conditions such as trophoblastic diseases, chorionic epithelioma, hydatic mole and certain non-trophoblastic neoplasms can cause the level of hCG to be elevated and cause false positive result. These diagnoses should be considered if consistent with the clinical evidence. Extra ulterine pregnancy, toxemia of pregnancy or threatened abo rtion can cause the hCG excretion decrease and lead to false negative result. The agglutination of the reagents also can be interfered by the usage of soaps and detergents.

A pregnancy test is used to determine if a woman is pregnant. This is done by analysing a sample of a woman's urine or blood for the presence of human chorionic gonadotropin (hCG). hCG is a hormone produced by the placenta and is usually only present in a woman's body when she is pregnant. A pregnancy test is usually performed wh en a health professional or a woman suspects she is pregnant, or before using clinical services which may interfere with a pregnancy (e.g. prescription of hormonal contraceptives). Pregnancy tests are also used for other, less common reasons, such as diagnosing abnormally progressing pregnancies and gestational trophoblastic disease (molar pregnancy). Conclusion From the result of test, the sample and the Negative pregnancy control is negative and shows no agglutination meanwhile for the Positive pregnancy control shows positive result because the agglutination occur due to the reaction between the hCG and the latex reagnt coated with anti-hCG antibodies.

 
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References : 1. Burtis C.A (2001). Tietz Fundamentals of Clinical Chemistry 5th Edition. Elsevier Saunders. 2. Pregnancy Test. Retrieved from http://en.wikipedia.org/wiki/Pregnancy_test on 19th Feb 2011. 3. Understanding pregnancy. Retrieved from http://www.nlm.nih.gov/medlineplus/prenataltesting.h tml on 19th Feb 2011