Pregnancy tests determine pregnancy by detecting a hormone in your system that only pregnant women naturally posess (though false positives are not rare due to drugs and possibly even diet). This hormone is Human Chorionic Gonadtropin (hCG). It is secreted by the placenta after fertilization. Since the test is for a hormone, accurate testing is done best in the morning when there is the highest potency of the hormone, and the presence can be dilluted if the body is flushed with liquids.
The tests work by binding the hCG hormone, from either blood or urine, to an antibody and an indicator. The antibody will only bind to hCG; other hormones will not give a positive test result. The usual indicator is a pigment molecule, present in a line across a home pregnancy urine test.
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