Isolation And Identification Of Pathogenic Bacteria Of Mastitis And Establishment Detection Method Of Quadruple PCR By Buffalo

In this study,8 8 milk samples form Buffalo Institute in Guangxi wer e collected,isolation and identification the Pathogenic bacteria,a rapid quantit ativeanalytical method of quadruple PCR was established accoring to the m ain pathogenic bacteria species of buffalo mastitis,optimized and to evalua te its Clinical detection.Experiment content:1?112 strains of pathogenic bacteria were isolated from 56 milk samples by bacterial isolation and identification of 88 milk samples,Including 47 strains of Staphylococcus,41.9%;Streptococcus 25 strains,22.3%;21 strains of intestinal bacteria,18.8%;12fungal strains,10.8%;other bacteria 7 strains,6.2%.2?Staphylococcus aureus NUC ge ne,Escherichia coli rrrnb gene,Streptococcus lactis EF-Tu gene,Staphyloc occus epidermidis se2313 sequence Design and choose four pairs of specif ic primers,established and optimized a fast diagnostic method of quadru ple PCR for simultaneous detection the 4 pathogenic bacterias.Experimental results:1?The resulting in Buffalo mastitis main pathoge ns were Staphylococcus aureus,Staphylococcus aureus,Streptococcus lactis and Escherichia coli,and part of the fungal infection;mainly is given pr iority to mixed infection,in 55 samples of milk samples there are 40 hav e two and above variety of pathogenic bacteria mixed infection,accounted for 71.4%.2?PCR products were electrophoresis,the Escherichia coli,S taphylococcus aureus,Staphylococcus epidermidis,streptococcus agalactiae respectively in 722bp,279bp,581bp and 197bp have amplified correspond ing bands and no other non-specific products,experimental and clinical de tection results show that the method has high Sensitivity and specificity.Compared with the traditional detection method of bacteria,the result is a ccurate,and greatly shorten the detection time and improve the detection efficiency.