Dairy cow milk progesterone colloidal gold test paper strip and preparation method thereof
A technology for detecting test strips and colloidal gold, applied in the field of biochemistry, can solve problems such as the uncertainty of rectal examination and the high price of B-ultrasound equipment
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[0071] The present invention is achieved in this way, the preparation method of a kind of dairy cow milk progesterone colloidal gold detection test paper strip comprises:
[0072] S101: Preparation of main solutions, including preparation of trisodium citrate solution, preparation of chloroauric acid solution, preparation of K 2 CO 3 Solution, preparation of PBS solution, preparation of BSA solution, preparation of sample pad treatment solution, preparation of gold standard complex solution;
[0073] S102: Preparation and identification of colloidal gold, including preparation of colloidal gold, quality identification of colloidal gold, preparation and purification of gold-labeled progesterone antibody complex, selection of antigen coating solution and gold-labeled diluent, and selection of gold-labeled antibody diluent and dilution selection, NC membrane selection and secondary antibody coating concentration selection, NC membrane blocking solution and blocking time selectio...
Embodiment 1
[0131] Determination of various properties of progesterone colloidal gold test strips
[0132] Milk sample test: Different batches of test strips were used to test the milk samples of 25 pregnant and 25 non-pregnant dairy cows confirmed by rectal method.
[0133] Determination of the detection limit of test strips: Progesterone standard solutions of different concentrations (concentrations of 1ng / mL, 3ng / mL, 5ng / mL, 7ng / mL, 9ng / mL, 11ng / mL, 13ng / mL, 15ng / mL , 17ng / mL), dissolved in methanol. After the test strips were assembled, 50 μL of progesterone standard solutions of different concentrations were added dropwise to the test strips, and after standing at room temperature for 5 minutes, PBS was used as a negative control to observe the test results.
[0134] Specificity test: 11-α-hydroxyprogesterone, estradiol, testosterone, follicle-stimulating hormone, estrogen, androstenedione, cortisol, and corticosterone were prepared in PBS to a standard concentration solution of 100...
Embodiment 2
[0176] Embodiment two, the clinical application of test two colloidal gold test strips
[0177] In this embodiment, the standard intensive cattle farm in the eastern part of Heilongjiang is selected for the clinical application of progesterone colloidal gold test strips. The test strips are mainly used to detect the pregnancy of dairy cows, and the performance of the test strips is improved through the test results. The relationship between progesterone content in the body and certain reproductive disorders.
[0178] Materials and Methods
[0179] 1.1 Main Instruments
[0180] The centrifuge Anke TDL80-2B was purchased from Shanghai Anting Scientific Instrument Company; the ultra-low temperature refrigerator; the T6 UV-Vis spectrophotometer was purchased from Beijing General Instrument Co., Ltd.; the incubator was purchased from Shanghai Yuejin Medical Devices No. 1 Factory.
[0181] 1.2 Main detection reagents
[0182] Progesterone ELISA kit was purchased from Kamishu Sha...
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