沙门氏菌invH基因的序列分析与分子检测

DNA Sequence Analysis and Molecular Detection of invH Gene from Salmonella spp.

根据沙门氏菌的invH基因序列设计1对引物,应用聚合酶链反应技术,分别对3种沙门氏菌标准菌株和5种非沙门氏菌株进行PCR扩增,结果3种标准沙门氏菌株均扩增出479 bp的特异条带,非沙门氏菌皆无特异带扩增。对3种标准株的扩增片段进行克隆及序列分析,证实沙门氏菌的invH基因序列比较保守;对19种地方分离沙门氏菌进行PCR特异性检测,结果有16株扩出特异性条带,表明本研究建立的沙门氏菌检测方法具有较高的特异性。

A pair of oligonucleotide primers were synthesized according to the invH gene nucleotide sequence of Salmonella spp. and the invH gene was amplified from 3 strains of Salmonella and 5 strains of no - salmonella bacteria by PCR. The results showed that 3 strains of Salmonella yielded the 479 bp specific amplicon, but no specific product yielded in no - salmonella bacteria. The PCR products of 3 Salmonella spp. were cloned and sequenced. The result showed that the DNA sequences of invH gene from Salmonella spp. were consistent with the previous report. The invH gene - based PCR method for detecting Salmonella spp. was used to detect Salmonella spp. of 19 districts. 16 specific strains were amplified. It is concluded that the invH gene -based PCR method is highly specific.