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

Inactive Publication Date: 2013-02-27
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to solve the problems of ELISA and IHA diagnostic methods for bovine schistosomiasis japonicum, which have many operating steps, slow speed, and cannot meet the needs of on-site quarantine. and other defects, provide a sensitive, specific, safe, stable, high signal strength, solid state, non-polluting detection test strip for the diagnosis, quarantine and census of bovine schistosomiasis japonicum, and it can be tested within 5 to 15 minutes. Fast and easy detection method that can be completed within

Method used

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  • Preparation of test strip for detection of antibody of bovine schistosoma japonicum katsurada and application method thereof
  • Preparation of test strip for detection of antibody of bovine schistosoma japonicum katsurada and application method thereof
  • Preparation of test strip for detection of antibody of bovine schistosoma japonicum katsurada and application method thereof

Examples

Experimental program
Comparison scheme
Effect test

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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Abstract

The invention discloses preparation of a test strip for detection of antibody of bovine schistosoma japonicum katsurada and an application method thereof, which belong to the technical field of diagnosis of animal parasitic diseases. The test strip consists of a sample gasket, an immune probe dry plate, a chromatographic film, water absorbing paper, a PVC (Polyvinyl Chloride) base plate, and an arrow label coating and a handle label coating which are combined. The immune probe dry plate is prepared by forming a covalent bond conjugate by carboxyl of polystyrene latex beads and amido of rabbit anti-bovine IgG antibody protein and absorbing on a polyester film. The chromatographic film is prepared by using a schistosome soluble antigen anchored on a nitrocellulose membrane as a detection line and goat anti-rabbit IgG antibody as a quality control line. The schistosome antibody in bovine serum can be quickly detected. The method is simple and convenient to operate, and the results are intuitional and accurate. The method can be widely applied to diagnosis, general survey and quarantine of bovine schistosome.

Description

【Technical field】 [0001] The invention relates to the technical field of diagnosis of livestock parasitic diseases, in particular to the preparation and application method of bovine schistosomiasis antibody detection test strips. 【Background technique】 [0002] Schistosomiasis is a zoonotic parasitic disease. At present, there are still more than 100 counties in 7 provinces (autonomous regions) in my country that have not reached the transmission control standards for schistosomiasis. Cattle are an important source of infection for schistosomiasis. Due to natural factors such as seasonal floods in the Yangtze River and social factors such as frequent movement of humans and animals, the epidemic of schistosomiasis in my country has recurred in recent years, and the number of sick humans and animals has increased. my country has always listed schistosomiasis as an important object of animal disease surveillance and quarantine. Therefore, the transit quarantine of schistosom...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/531
Inventor 卢福庄张雪娟顾昀付媛季敬余冯尚连俞国乔杨玉焕周煜阳爱国董国栋郭莉毛光琼石团员
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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