Enzyme immunoassay method for progestational hormone in excrement of sows and method for detecting oestrous cycle of sow

Abstract

The invention discloses an enzyme immunoassay method for progestational hormone in excrement of sows. The method comprises the steps of: (1) sample collection: collecting the excrement of the sows, weighing and dissolving 0.5 g of excrement of the sows into 20 ml of measuring buffer liquid, carrying out centrifuging, taking and placing supernatant at 4 DEG C to be detected; and (2) measuring content of progestational hormone in a sample by utilizing competitive ELISA (enzyme-linked immuno sorbent assay): measuring according to an ELISA process. According to the enzyme immunoassay method for the progestational hormone in the excrement of the sows, the detection sensitivity of progesterone reaches 0.2 ng / ml or 8 ng / g; the recovery rate of the progesterone is 95.78%; inter-assay and intra-assay variable coefficients are respectively 6.8% (n is equal to 9) and 11.23% (n is equal to 8); the specificity of the progesterone is that the cross reaction rates of the progesterone to estradiol, estriol and oestrone are all less than 0.01%, and the cross reaction rate of the progesterone to testosterone is 0.35%; the stability is good; the required sample is easy to collect, animals are not injured, a measured result is reliable, and a method for measuring blood of the animals can be replaced to monitor a series of reproductive activities of the animals.

Description

technical field [0001] The invention relates to an enzyme immunoassay method for progesterone in feces of sows, which measures the concentration of progesterone in feces to monitor the ovarian activity of sows, and belongs to the field of biotechnology. Background technique [0002] The principle of enzyme-linked immunosorbent assay (ELISA): The principle of competitive ELISA is to coat the antibody on a solid phase first, and then add a certain sample to be tested (including the antigen to be tested) and a certain amount of enzyme-labeled antigen conjugate. When they can competitively bind to the solid-phase antibody, after incubation and washing, the substrate is added to develop color, and their OD is determined 492 The content of the analyte was calculated according to the standard curve. If the content of progesterone in the sample is high, most of the progesterone antibodies coated on the solid phase carrier are bound by the free progesterone in the sample. The proge...

AI Technical Summary

Problems solved by technology

In the production process, the ELISA method is usually used to measure the estrogen / progesterone content of the collected animal blood samples, but this sampling method will cause certain stress to the animals, and it is difficult to implement

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