鸡白痢沙门氏菌直接-多重PCR检测体系的建立

Establishment of a direct multiplex PCR detection system of Salmonella pullorum

摘要:

通过对鸡白痢沙门氏菌毒力质粒pSPUV上入侵质粒抗原J蛋白和质粒共转移调控因子的编码基因 ipaJ 和 traJ 的序列分析,在特异区域设计PCR引物,经沙门氏菌属不同种及常见肠道致病菌的交叉验证,筛选出三对鸡白痢沙门氏菌特异检测引物ipaJ-417、traJ-387、traJ-476. 并且调试出一种利用两种DNA聚合酶的直接三重PCR检测体系:invA-211/traJ-387/16S-495,可以对鸡白痢沙门氏菌进行快速准确的鉴定.

Abstract:

In this paper, we screened out three pairs of specific detection primers of Salmonella pullorum, ipaJ417, traJ-387, and traJ-476, by the blast analysis of the coding genes ipaJ and traJ which coded the invasion plasmid antigen protein J and plasmid co-transfer regulatory factor in Salmonella pullorum virulence plasmid Pspuv. And these PCR primers were designed in special regions and identificated in the different species of Salmonella and common intestinal pathogens. And a common reaction buffer of two kinds of DNA polymerase for the direct triple PCR system was also established (invA-211/traJ-387/16S-495). This direct triple PCR system can be used for fast and accurate identification of Salmonella pullorum.