A 16-unit nucleic acid typing kit for bovine mastitis pathogenic bacteria and its detection method

Abstract

The invention provides a 16-membered bovine mastitis pathogenic bacterium nucleic acid typing kit and a detecting method thereof. the kit comprises a reagent A containing a 5*PCR buffer solution, BSA,DMSO, MgCl2, dNTP, dUTP, MgCl2, glycerol, primers and probes, a reagent B containing a UDG enzyme, an ExTaq enzyme and an enzyme storage solution, a reagent C containing deionized water, a positive quality control product which is a mixed liquid of recombinant plasmids of 15 pathogenic bacteria and a beta-lactamase amplification sequence, wherein the mixed liquid is obtained through uniformly mixing mother liquids of 16 detected objects through TE buffer, and a negative quality control product which is a TE buffer solution. The cow mastitis PCR detection method is based on a multiple TaqMan probe real-time fluorescent PCR technology, realizes parallel detection of 15 pathogenic bacteria and a beta-lactamase resistance gene, has operation time of 4h and solves the problem that the traditional cow pathogenic bacterium detection method has low specificity, low sensitivity, complex processes and large time consumption and the existing detection method of pathogenic bacteria is not comprehensive.

Description

technical field [0001] The invention belongs to the technical field of detection of cow mastitis pathogenic bacteria, and relates to a 16-unit bovine mastitis pathogenic bacteria nucleic acid typing kit and a detection method thereof. Background technique [0002] Dairy cow mastitis (mastitis) is the inflammation of dairy cow mammary glands caused by stimulation of physical, chemical, microbial and other factors. One of the four major diseases in China, seriously affecting the development of the dairy industry. [0003] According to the statistics of the World Dairy Cattle Association, there are currently about 220 million dairy cows in the world, of which about 30% suffer from various types of mastitis. According to whether there are clinically visible changes in the udder of affected dairy cows, the National Mastitis Committee (NMC) of the United States divides them into clinical mastitis (Clinical Mastitis) and subclinical mastitis (Subclinical Mastit.is). The incidence...

AI Technical Summary

Problems solved by technology

However, domestic related patents mainly detect several pathogenic bacteria that cause mastitis in dairy cows. There are too few types of detection, and only for pathogenic bacteria, the detection range is limited.

[0007] At present, the detection of the pathogenic bacteria of dairy cow mastitis and its drug resistance is mainly through bacterial isolation and drug sensitivity test, which takes a long time and the accuracy of the results is not high

However, domestic related patents mainly detect several pathogenic bacteria that cause mastitis in dairy cows. There are too few types of detection, and ordinary PCR methods are mostly used, and only for pathogenic bacteria. There is no method for simultaneous detection of pathogenic bacteria and resistance genes.

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