Method for rapidly detecting buffalo fasciola spp antibody

A detection method and buffalo technology, applied in the field of cattle blight detection, can solve the problems of high technical requirements for operation, unsuitable for popularization and use, time-consuming, etc., achieving good application prospects, promoting monitoring and purification work, and rapid detection.

Active Publication Date: 2019-03-22
GUANGXI UNIV
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Problems solved by technology

[0004] At present, for the detection of fascioliasis in buffaloes, the application methods mainly include polymerase chain reaction (PCR), egg detection method, indirect ELISA method, etc., but these detection methods have high requirements for personnel and special equipment. Equipment, and time-consuming and other reasons, are not suitable for promotion and use at the grassroots level

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  • Method for rapidly detecting buffalo fasciola spp antibody
  • Method for rapidly detecting buffalo fasciola spp antibody
  • Method for rapidly detecting buffalo fasciola spp antibody

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Embodiment

[0044] Early preparation:

[0045] Materials: bovine serum albumin (BSA), tetrachloroauric acid (Sigma), analytically pure trisodium citrate, goat anti-mouse IgG secondary antibody, 135 nitrocellulose membrane (NC membrane), glass fiber paper OB6, ordinary absorbent paper, PVC bottom plate.

[0046] 1. Anti-buffalo IgG monoclonal antibody 2G7

[0047] 1. Purification of anti-buffalo IgG monoclonal antibody 2G7, the specific steps are as follows:

[0048] 1) Take the ascites produced by injecting the hybridoma cell line 2G7 into BaLb / C mice, and centrifuge at 4°C and 12000rpm for 15min to remove impurities;

[0049]2) Mix acetate buffer solution with mouse ascites at a ratio of 2:1, stir at room temperature, and add n-octanoic acid dropwise (33 μL n-octanoic acid per ml of ascitic fluid), mix at room temperature for 30 minutes to obtain a mixture ;

[0050] 3) Let the mixed solution stand at 4°C for at least 2 hours to allow the mixed solution to fully precipitate;

[0051...

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Abstract

The invention discloses a method for rapidly detecting a buffalo fasciola spp antibody, and belongs to the technical field of cattle plague disease detection. The method comprises the following steps:S1, preparation of an anti-buffalo IgG (Immunoglobulin G) monoclonal antibody 2G7 golden standard antibody conjugate pad; S2, preparation of a detection band: spraying a goat anti-mouse IgG antibodyand a fasciola spp ES antigen on a nitrocellulose membrane respectively to serve as a line C and a line T; S3, assembly of a detection strip; and S4, detection and result judgement: judging whether the antibody is positive or negative through a color development reaction of the detection band. The method has the advantages of rapid detection, easiness and convenience in operation, easiness in judgement, high specificity and the like, and provides a practical and effective method for detecting the buffalo fasciola spp antibody. The method has low requirements on operation techniques of detection personnel, does not need any special instrument equipment, and is suitable for being used during popularization on grass roots.

Description

technical field [0001] The invention relates to the technical field of bovine epidemic disease detection, in particular to a rapid detection method for buffalo fasciola antibodies. Background technique [0002] Fasciola spp is one of the most important parasitic diseases infecting ruminants such as cattle and sheep, and it is also a foodborne zoonotic parasitic disease. The large fasciola that parasitizes in the liver bile duct of cattle can cause acute or chronic hepatitis and cholangitis, and secondary systemic poisoning and nutritional disorders, often causing a large number of calves to die. The fascioliasis of cattle is mostly chronic, and the clinical symptoms vary according to the infection intensity, age of cattle, feeding management conditions, resistance and other factors. In mild infections, except for calves (1.5 to 2 years old) who are symptomatic, affected animals are often asymptomatic, making the disease difficult to detect in adult herds. However, if a lar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/543G01N33/577G01N33/535
CPCG01N33/535G01N33/54326G01N33/577
Inventor 吴文德张为宇侯林静
Owner GUANGXI UNIV
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