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Separation and identification method of cow mastitis pathogens

A cow mastitis and identification method technology, which is applied in the field of dairy cow mastitis prevention and treatment, can solve the problems of poor polymorphism of SSCP spectrum and the difference of other pathogenic bacteria, and achieve the effect of simple and convenient identification process, easy to master and simple operation

Inactive Publication Date: 2014-03-19
ANHUI NORMAL UNIV
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Problems solved by technology

However, some pathogenic bacteria have poor polymorphisms in the SSCP spectrum, making it difficult to distinguish them from other pathogenic bacteria.

Method used

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  • Separation and identification method of cow mastitis pathogens
  • Separation and identification method of cow mastitis pathogens
  • Separation and identification method of cow mastitis pathogens

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Experimental program
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Embodiment Construction

[0052] The present invention will be described in detail below according to the accompanying drawings, which is a preferred embodiment among various implementations of the present invention.

[0053] (1) Milk sample collection: 40 milk samples were collected from 40 dairy cows with mastitis in Weigang Dairy Farm, Wuhu, Anhui.

[0054] (2) Isolation of pathogenic bacteria: the collected milk samples were inoculated into ordinary medium, MacConkey agar medium, and beef extract peptone medium respectively. Cultivate at 37°C for 24-48 hours, and perform purification according to the colony morphology. Smear, Gram stain, and microscopic examination were performed according to conventional methods. Preliminary identification of pathogenic bacteria species.

[0055](3) DNA extraction of pathogenic bacterial colonies: Pick an appropriate amount of colonies into a centrifuge tube filled with 1000 μl sterile water, and centrifuge at 12000 r / min for 5 min. The water was blotted off to...

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Abstract

The invention relates to a separation and identification method of cow mastitis pathogens. Separation and identification on the Chinese Holstein cow mastitis pathogens are carried out by adopting a traditional bacterial culture method and a sequence analysis method and a double-primer polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) technology on the basis of pathogen 16S rRNA (ribosomal Ribonucleic Acid) genes. The identification result of the double-primer PCR-SSCP technology is basically the same as that of the sequence analysis method. The result of the double-primer PCR-SSCP technology is good in polymorphism, and is similar to the sequence analysis result. The separation and identification method is an efficient, simple and economic method for identifying the pathogens, and a theoretical foundation is provided for rapid identification of the Chinese Holstein cow mastitis pathogens. Thus, a theoretical basis is provided for diagnosis and treatment of cow mastitis.

Description

technical field [0001] The invention relates to the field of prevention and treatment of dairy cow mastitis, in particular to using double primer PCR-SSCP of 16S rRNA gene as an efficient, simple and economical method to identify the pathogenic bacteria of cow mastitis. Background technique [0002] Cow mastitis is one of the most serious diseases in dairy farming worldwide. Mastitis can have a serious impact on milk production and quality, consuming an estimated 38% of all disease economies. Under the stimulation of physical and chemical factors, breast tissue will produce inflammation, but it is mainly caused by one or more microorganisms infecting the mammary gland. According to reports, mastitis is mainly due to bacterial infection, and there are a few cases due to fungal infection. Staphylococcus aureus, for example, often causes gangrenous mastitis in dairy cows. At present, there are many ways to prevent and treat mastitis in dairy cows, mainly through optimized ma...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04C12R1/01
CPCC12Q1/6869C12Q1/686C12Q2531/113C12Q2565/131C12Q2537/143
Inventor 杨兰英蔡亚非刘再群
Owner ANHUI NORMAL UNIV
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