Method for quickly detecting pathogenic bacteria in cow endometritis

A technology for endometritis and pathogenic bacteria, applied in the field of microbial detection, can solve the problems of difficult detection of microorganisms and easy co-migration, and achieve the effects of sensitive and accurate results, fast detection and low cost

Active Publication Date: 2019-05-03
NORTHWEST A & F UNIV
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  • Claims
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Problems solved by technology

PCR-DGGE technology can analyze a large number of samples at the same time, quickly and accurately identify individual microorganisms, and has the advantages of high resolution and small sample demand, but this technology a

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  • Method for quickly detecting pathogenic bacteria in cow endometritis
  • Method for quickly detecting pathogenic bacteria in cow endometritis
  • Method for quickly detecting pathogenic bacteria in cow endometritis

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Embodiment 1

[0027] Embodiment 1 A method for rapidly detecting pathogenic bacteria in cow endometritis, specifically comprising the following steps:

[0028] 1) Extraction of bacterial genomic DNA: Escherichia coli (E.coli), Staphylococcus aureus (S.aureus), Cryptobacter pyogenes (T.pyogenes), DNA from S. uberis, P. mirabilis, P. aeruginosa.

[0029] 2) Multiplex PCR: In the reaction system containing the above-mentioned bacterial DNA, a PCR primer system capable of simultaneously detecting 6 kinds of bacterial specific sequences is added for amplification.

[0030] According to the ycjM gene of Escherichia coli, the thermostable nuclease gene nuc of Staphylococcus aureus, the hemolysin plo gene of pathogenic Cryptobacter pyogenes, the plasminogen kinase pauA gene of Streptococcus uberis, and the urease synthesis of Proteus mirabilis Primers were designed for the positive regulator R gene (ureR) and the O antigen acetyltransferase gene of Pseudomonas aeruginosa.

[0031] The designed 6 ...

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Abstract

The invention discloses a method for quickly detecting pathogenic bacteria in cow endometritis. The method comprises the following steps of (1) extracting DNA of 6 common pathogenic bacteria which arecultured over night, in womb secretion by a reagent kit method; (2) in a reaction system containing the DNA of the bacteria, adding a PCR primer system which can detect the specificity sequence of the 6 kinds of bacteria at the same time, and performing amplification; and (3) preparing 2% of agarose gel, performing gelatin electrophoresis, wherein the voltage is 120 V and the time is 45 min, andthen performing electrophoresis ultraviolet imaging. Based on a multiple polymerase chain reaction technique, the method disclosed by the invention can realize detection of 6 kinds of the bacteria, including E.coli, S.aureus, T.pyogenes, S.uberis, P.mirabilis and P.aeruginosa. Compared with sequencing, techniques of substance PCR and the like are simpler and more convenient to operate, lower in cost, higher in sensitivity and specificity. The detection and identification period for clinical bacterium detection are selected.

Description

technical field [0001] The invention belongs to the field of microorganism detection, in particular to a method for rapidly detecting pathogenic bacteria in cow endometritis. Background technique [0002] Cow endometritis (endometritis) is a disease of cows caused by pathogenic microorganism infection and other factors to cause inflammation of the endometrium, resulting in pathological changes in the endometrium layer. Most of the disease occurs during parturition or postpartum of dairy cows, and it is the most common reproductive disorder of dairy cows. Infertility caused by endometritis can account for 60.0%-70.0% of disease-induced infertility. The disease can lead to repeated infertility and even death of affected cows, which has a long-term irreversible impact on the reproductive system of dairy cows, resulting in increased feeding costs, decreased milk production, and even increased culling rates, which have brought serious problems to dairy farms. serious economic lo...

Claims

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Application Information

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IPC IPC(8): C12Q1/689C12Q1/14C12Q1/10C12Q1/04C12R1/19C12R1/445C12R1/465C12R1/385C12R1/37C12R1/01
Inventor 李勤凡王宁宁王晶范玉堂许信刚
Owner NORTHWEST A & F UNIV
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