Method for the determination of steroid metabolite content in sheep fecal excrement
A technology of steroids and metabolites, applied in the field of scientific research and experimentation, can solve problems such as time-consuming, little research on domesticated animals, and carcass damage
Inactive Publication Date: 2017-02-08
LINYI UNIVERSITY
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Problems solved by technology
[0002] At present, studies on the determination of steroid metabolites in feces mainly focus on wild animals and rare birds, and few studies have been done on domestic animals.
However, the collection of feces and excreta of wild animals and rare birds requires blood collection or tissue damage sampling, which will cause damage to the carcass
[0003] At present, the determination of steroid metabolites in feces is mainly used in kits, which is expensive and time-consuming
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[0060] The present invention will be further described in detail below in conjunction with specific embodiments.
[0061] 1. Reagent preparation
[0062] 1.1 The first coating solution: Na 2 CO 3 1.59g, NaHCO 3 2.93g, dissolved in double distilled water, adjusted to pH 9.6;
[0063] 1.21mol / LHCl;
[0064] 1.3 Buffer: 2.42g Tris, 20mmol / L; 17.9g NaCl, 0.3mol / L; 1g bovine serum albumin; 1ml Tween 80; pH adjusted to 7.5;
[0065] 1.4 The second coating solution: 3.146g trihydroxyaminomethane; 23.3g NaCl; 13g BSA; 1.3g sodium azide, dissolved in double distilled water, and made up to 1.3L, and the pH was adjusted to 7.5.
[0066] 1.5 Cleaning solution: 0.5ml Tween 20, add 2.5L double distilled water.
[0067] 1.6 Horseradish peroxidase substrate buffer: 1.36g sodium acetate=10mmol / l, dissolve with double distilled water, and make 1L, adjust the pH to 5.0 with about 8ml of 5% citric acid.
[0068] 1.7 Enzyme concentrated solution for streptavidin reaction: 30ml buffer + 0.001ml streptavidin-hor...
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The invention relates to the technical field of scientific research test, and in particular relates to a method for measuring the content of steroid metabolites in dung excrements of sheep. A self-prepared reagent is adopted for measuring the content of the steroid metabolites in dung. The method comprises the measurement and analysis steps of (1) preparing the reagent; (2) covering a microporous plate with proteins A; (3) preparing a storage solution; (4) preparing working liquid; (5) analyzing; and (6) measuring and calculating the absorbency. The method for measuring the content of the steroid metabolites in the dung excrements of the sheep by adopting the self-prepared reagent is operated mainly based on the self-prepared reagent in a lab, so that the cost is lowered; furthermore, the content of the steroid metabolites can be accurately measured. According to the method for measuring the content of the steroid metabolites in the dung excrements of the sheep, non-tissue damage sampling is realized, so that the influence on the content of the steroid metabolites caused by blood sampling is avoided, and injury to livestock is avoided.
Description
Technical field [0001] The invention relates to the field of scientific research and experimentation, in particular to a method for determining the content of steroid metabolites in sheep feces. Background technique [0002] At present, researches on the determination of steroid metabolites in fecal excretion mainly focus on wild animals and rare birds, and there are very few studies in domestic animals. The collection of feces and excrements of wild animals and rare birds requires blood sampling or tissue damage sampling, which can cause damage to the carcass. [0003] At present, the determination of steroid metabolites in stool is mainly used in kits, which is expensive and time-consuming. Summary of the invention [0004] In order to solve the shortcomings in the above technical problems, the present invention provides a method for determining the content of steroid metabolites in sheep feces excrement. The method is mainly based on the operation of self-prepared reagents in th...
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IPC IPC(8): G01N33/92G01N33/531
CPCG01N33/74G01N2333/723
Inventor 吕慎金杨燕
Owner LINYI UNIVERSITY