A rapid detection method for antibodies against Fasciola macula in buffalo
A detection method and buffalo technology, applied in the field of cattle blight detection, can solve problems such as time-consuming, unsuitable for popularization and use, and high operational technical requirements, and achieve the effects of rapid detection, promotion of monitoring and purification work, and good application prospects.
Active Publication Date: 2022-02-01
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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[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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The invention discloses a rapid detection method for buffalo fasciola antibodies, and belongs to the technical field of bovine epidemic detection. The invention comprises the following steps: S1, preparation of anti-buffalo IgG monoclonal antibody 2G7 gold-labeled antibody binding pad; S2, preparation of detection zone, spraying sheep anti-mouse IgG antibody and Fasciola ES antigen on the nitrocellulose membrane respectively , as C line and T line; S3, assembly of the test strip; S4, detection and result judgment, judged as positive or negative by the color reaction of the test strip. The invention has the advantages of rapid detection, simple and convenient operation, easy determination, strong specificity, etc., and can provide a practical and effective method for the detection of buffalo fasciola. This method does not require high technical requirements for testing personnel, and does not require special instruments and equipment, and is suitable for popularization and use at the grassroots level.
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...
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IPC IPC(8): G01N33/543G01N33/577G01N33/535
CPCG01N33/535G01N33/54326G01N33/577
Inventor 吴文德张为宇侯林静
Owner GUANGXI UNIV