Preparation of test strip for detection of antibody of bovine schistosoma japonicum katsurada and application method thereof
A technology for schistosomiasis and test strips, which is applied in the field of preparation of bovine schistosomiasis antibody detection test strips, which can solve the problems of inability to meet on-site quarantine requirements, short storage period, and inability to transport by air or mail
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Embodiment 1
[0055] Example 1: (Preparation 1 of test strip for detecting bovine schistosomiasis antibody)
[0056] 1 Preparation of test strips
[0057] 1.1 Preparation of immunoprobe dry film
[0058] 1.1.1 Preparation of rabbit anti-yellow buffalo IgG
[0059] ⑴ Extraction of yellow buffalo IgG
[0060] ① Aseptically collect the blood of healthy cattle and buffalo, and separate the serum respectively.
[0061] ② Take 5ml of cattle and buffalo serum and mix them, add 10ml of 0.85% NaCl and mix evenly, add 8.6ml of saturated ammonium sulfate (SAS) drop by drop to make it reach 30% saturation, stir while adding, and adjust the pH value with concentrated ammonia water to 7.0, put it at 4°C overnight, take it out to room temperature for 1hr, centrifuge at 3000r / min for 30min, discard the supernatant, add 20ml of 0.85% NaCl to dissolve the precipitate. Then add 10.8ml of SAS drop by drop to make it reach 35% saturation, stir while adding, adjust the pH value to 7.0 with concentrated ammonia...
Embodiment 2
[0172] Example 2: (Preparation 2 of the test strip for detecting bovine Schistosomiasis japonicum antibody)
[0173] 1 Production of test strips
[0174] (1) The preparation of the immune probe dry sheet is the same as that in 1.1 in Example 1, but there are 5 differences:
[0175] ① Use 300nm blue latex microspheres.
[0176] ②The rabbit anti-yellow buffalo IgG protein content used in latex microsphere labeling is 2mg / ml.
[0177] ③Preparation of blocking solution Weigh 0.500g of casein, 10.000g of sucrose, NaN 3 All 0.020g was put into 100ml pH7.20.05M Tris-HCl buffer solution, heated in a 37-40°C water bath until completely dissolved. That is, it is formulated to contain 0.5% casein, 10% sucrose, NaN 3 Blocking solution composed of 0.02% pH7.20.05M Tris-HCl buffer.
[0178] ④Preparation of polyester film treatment solution: 0.2M Tris 125ml, 0.1N HCl 225ml, casein 2.500g, sucrose 50.000g, NaN 3 Mix 0.100g, dissolve in 37°C water bath, then add Tween-201.5ml and stir ev...
Embodiment 3
[0187] Example 3: (Preparation 3 of the test strip for detecting bovine Schistosomiasis japonicum antibody)
[0188] 1 Production of test strips
[0189] (1) The preparation of the immune probe dry sheet is the same as that in 1.1 in Example 1, but there are 4 differences:
[0190] ①The rabbit anti-yellow buffalo IgG protein content used in latex microsphere labeling is 3mg / ml.
[0191] ②Preparation of blocking solution Weigh 0.300g of casein, 5.000g of sucrose, NaN 3 All 0.030g was put into 100ml pH7.20.05M Tris-HCl buffer solution, heated in a 37-40°C water bath until completely dissolved. That is, it is formulated to contain 0.3% casein, 5% sucrose, NaN 3 Blocking solution composed of 0.03% pH7.20.05M Tris-HCl buffer.
[0192] ③Preparation of polyester film treatment solution: 0.2M Tris.125ml, 0.1N HCl 225ml, casein 1.500g, sucrose 25.000g, NaN 3 Mix 0.150g, dissolve in a water bath at 37°C, then add Tween-202.0ml and stir evenly, add distilled water to 500ml and mix e...
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