畜禽源3种革兰氏阴性杆菌多重PCR检测方法的建立与应用

doi:10.16431/j.cnki.1671-7236.2016.03.003

Abstract

Abstract: To establish a rapid and accurate multiplex PCR method for the clinical detection of three main Gram-negative bacilli from livestock and poultry in Guizhou province,three pairs of specific primers were designed and synthetized according to 23S rRNA gene of Escherichia coli(E.coli),KMT gene of Pasteurella and invA gene of Salmonella.In this study,A mutiplex PCR reaction system was established for detecting the three pathogens at the same time,and the reaction system was optimized and it's performance was evaluated.The results showed that the best concentration of primers was 1.0,1.5and 1.0 μmol/L,respectively,and the best annealing temperature was 56 ℃.The results of performance evaluation showed that the method had good specificity and sensitivity,the sensitivity of Salmonella could reach to 1.44 pg/μL,was the highest.70 Gram-negative clinical isolated strains were detected by the multiplex PCR and it was proofed that the method could identify the three pathogens rapidly and accurately,and it could provide effective technical for the rapid clinical diagnosis and epidemiological survey.

Key words: multiplex PCR; E.coli; Pasteurella; Salmonella

CLC Number:

S852.61

MA Guang-qiang, LIU Li-juan, LONG Dan-dan, YUE Jun, LI Yi, WEN Ming, CHENG Zhen-tao, ZHOU Bi-jun. Establishment and Application of Multiplex PCR Detection Method for Three Main Gram-negative Bacilli from Livestock and Poultry[J]. , 2016, 43(3): 577-584.

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Establishment and Application of Multiplex PCR Detection Method for Three Main Gram-negative Bacilli from Livestock and Poultry

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