Development and evaluation of a multiplex real-time polymerase chain reaction procedure to clinically type prevalent Salmonella enterica serovars.

The Journal of Molecular Diagnostics : JMD
Nélida MúñozNora Cardona-Castro

Abstract

A multiplex real-time polymerase chain reaction procedure was developed to identify the most prevalent clinical isolates of Salmonella enterica subsp. enterica. Genes from the rfb, fliC, fljB, and viaB groups that encode the O, H, and Vi antigens were used to design 15 primer pairs and TaqMan probes specific for the genes rfbJ, wzx, fliC, fljB, wcdB, the sdf-l sequence, and invA, which was used as an internal amplification control. The primers and probes were variously combined into six sets. The first round of reactions used two of these sets to detect Salmonella O:4, O:9, O:7, O:8, and O:3,10 serogroups. Once the serogroups were identified, the results of a second round of reactions that used primers and probes for the flagellar antigen l genes, 1,2; e,h; g,m; d; e,n,x; and z(10), and the Vi gene were used to identify individual serovars. The procedure was standardized using 18 Salmonella reference strains and other enterobacteria. The procedure's reliability and sensitivity was evaluated using 267 randomly chosen serotyped Salmonella clinical isolates. The procedure had a sensitivity of 95.5% and was 100% specific. Thus, our technique is a quick, sensitive, reliable, and specific means of identifying S. enterica serovars and...Continue Reading

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Citations

Jun 24, 2011·Journal of Clinical Microbiology·Kristyn FranklinMohamed A Karmali
Sep 16, 2014·Diagnostic Microbiology and Infectious Disease·Catherine YoshidaRoderick M Card
May 21, 2013·Journal of Clinical Laboratory Analysis·José Luis Villarreal CamachoDaniel Antonio Villanueva Torregroza
Sep 29, 2012·Environmental Microbiology·Salim T Islam, Joseph S Lam
Nov 16, 2013·Critical Reviews in Microbiology·Chunlei ShiAndrea Isabel Moreno Switt
Mar 13, 2014·Journal of Microbiological Methods·Ligong ZhaiXiaomei Bie
May 14, 2017·Applied and Environmental Microbiology·Kuppuswamy N Kasturi, Tomas Drgon
Sep 6, 2018·Revista de salud pública·Elkin E Hernández-PorrasJaime Moreno
Dec 5, 2019·Frontiers in Cellular and Infection Microbiology·Minxu LiQinghua Hu
Mar 25, 2021·Biomédica : revista del Instituto Nacional de Salud·Nancy Yaneth FlórezJaime Enrique Moreno
Jul 4, 2020··Jingxuan Wei, Jingxuan Wei

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