Detection of Salmonella spp. Using a Generic and Differential FRET-PCR

文献类型： 外文期刊

第一作者： Zhang, Jilei

作者： Zhang, Jilei;Wei, Lanjing;Xu, Chuanling;Wang, Chengming;Zhang, Jilei;Wei, Lanjing;Gong, Jiansen;Xu, Bu;Pan, Zhiming;Xu, Chuanling;Wang, Chengming;Kelly, Patrick;Jaegerson, Kirsten;Freeman, Mark;Gong, Jiansen;Xu, Bu;Pan, Zhiming

作者机构：

期刊名称：PLOS ONE （ 影响因子：3.24； 五年影响因子：3.788 ）

ISSN： 1932-6203

年卷期： 2013 年 8 卷 10 期

页码：

收录情况： SCI

摘要： To facilitate the detection of Salmonella and to be able to rapidly and conveniently determine the species/subspecies present, we developed and tested a generic and differential FRET-PCR targeting their tetrathionate reductase response regulator gene. The differential pan-Salmonella FRET-PCR we developed successfully detected seven plasmids that contained partial sequences of S. bongori and the six S. enterica subspecies. The detection limit varied from similar to 5 copies of target gene/per PCR reaction for S. enterica enterica to similar to 200 for S. bongori. Melting curve analysis demonstrated a T-m of similar to 68 degrees C for S. enterica enterica, similar to 62.5 degrees C for S. enterica houtenae and S. enterica diarizonae, similar to 57 degrees C for S. enterica indica, and similar to 54 degrees C for S. bongori, S. enterica salamae and S. enterica arizonae. The differential pan-Salmonella FRET-PCR also detected and determined the subspecies of 4 reference strains and 47 Salmonella isolated from clinically ill birds or pigs. Finally, we found it could directly detect and differentiate Salmonella in feline (5/50 positive; 10%; one S. enterica salamae and 4 S. enterica enterica) and canine feces (15/114 positive; 13.2%; all S. enterica enterica). The differential pan-Salmonella FRET-PCR failed to react with 96 non-Salmonella bacterial strains. Our experiments show the differential pan-Salmonella FRET-PCR we developed is a rapid, sensitive and specific method to detect and differentiate Salmonella.

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