Detection of Salmonella spp. using a generic and differential FRET-PCR.

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 deve...

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Autores principales:	Jilei Zhang, Lanjing Wei, Patrick Kelly, Mark Freeman, Kirsten Jaegerson, Jiansen Gong, Bu Xu, Zhiming Pan, Chuanling Xu, Chengming Wang
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/d2f8e76d5c7d46828c732387f55fa504
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spelling	oai:doaj.org-article:d2f8e76d5c7d46828c732387f55fa5042021-11-18T08:50:51ZDetection of Salmonella spp. using a generic and differential FRET-PCR.1932-620310.1371/journal.pone.0076053https://doaj.org/article/d2f8e76d5c7d46828c732387f55fa5042013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24146814/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203To 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 ~5 copies of target gene/per PCR reaction for S. enterica enterica to ~200 for S. bongori. Melting curve analysis demonstrated a T m of ~68 °C for S. enterica enterica, ~62.5 °C for S. enterica houtenae and S. enterica diarizonae, ~57 °C for S. enterica indica, and ~54 °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.Jilei ZhangLanjing WeiPatrick KellyMark FreemanKirsten JaegersonJiansen GongBu XuZhiming PanChuanling XuChengming WangPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 10, p e76053 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Jilei Zhang Lanjing Wei Patrick Kelly Mark Freeman Kirsten Jaegerson Jiansen Gong Bu Xu Zhiming Pan Chuanling Xu Chengming Wang Detection of Salmonella spp. using a generic and differential FRET-PCR.
description	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 ~5 copies of target gene/per PCR reaction for S. enterica enterica to ~200 for S. bongori. Melting curve analysis demonstrated a T m of ~68 °C for S. enterica enterica, ~62.5 °C for S. enterica houtenae and S. enterica diarizonae, ~57 °C for S. enterica indica, and ~54 °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.
format	article
author	Jilei Zhang Lanjing Wei Patrick Kelly Mark Freeman Kirsten Jaegerson Jiansen Gong Bu Xu Zhiming Pan Chuanling Xu Chengming Wang
author_facet	Jilei Zhang Lanjing Wei Patrick Kelly Mark Freeman Kirsten Jaegerson Jiansen Gong Bu Xu Zhiming Pan Chuanling Xu Chengming Wang
author_sort	Jilei Zhang
title	Detection of Salmonella spp. using a generic and differential FRET-PCR.
title_short	Detection of Salmonella spp. using a generic and differential FRET-PCR.
title_full	Detection of Salmonella spp. using a generic and differential FRET-PCR.
title_fullStr	Detection of Salmonella spp. using a generic and differential FRET-PCR.
title_full_unstemmed	Detection of Salmonella spp. using a generic and differential FRET-PCR.
title_sort	detection of salmonella spp. using a generic and differential fret-pcr.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/d2f8e76d5c7d46828c732387f55fa504
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Detection of Salmonella spp. using a generic and differential FRET-PCR.

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