Real-Time PCR Method for Detection of Salmonella spp. in Environmental Samples

Applied and Environmental Microbiology
Kuppuswamy N Kasturi, Tomas Drgon

Abstract

The methods currently used for detecting Salmonella in environmental samples require 2 days to produce results and have limited sensitivity. Here, we describe the development and validation of a real-time PCR Salmonella screening method that produces results in 18 to 24 h. Primers and probes specific to the gene invA, group D, and Salmonella enterica serovar Enteritidis organisms were designed and evaluated for inclusivity and exclusivity using a panel of 329 Salmonella isolates representing 126 serovars and 22 non-Salmonella organisms. The invA- and group D-specific sets identified all the isolates accurately. The PCR method had 100% inclusivity and detected 1 to 2 copies of Salmonella DNA per reaction. Primers specific for Salmonella-differentiating fragment 1 (Sdf-1) in conjunction with the group D set had 100% inclusivity for 32 S Enteritidis isolates and 100% exclusivity for the 297 non-Enteritidis Salmonella isolates. Single-laboratory validation performed on 1,741 environmental samples demonstrated that the PCR method detected 55% more positives than the Vitek immunodiagnostic assay system (VIDAS) method. The PCR results correlated well with the culture results, and the method did not report any false-negative results. T...Continue Reading

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Citations

Aug 1, 2020·Frontiers in Microbiology·Carolina VillamilJohn Emerson Leguizamon
Oct 31, 2020·Brazilian Journal of Microbiology : [publication of the Brazilian Society for Microbiology]·Negar NarimisaFaramarz Masjedian Jazi
Dec 18, 2020·Frontiers in Microbiology·Luping XuArun K Bhunia
Jan 5, 2021·Biosensors & Bioelectronics·Elisa SantovitoDmitri B Papkovsky
Dec 22, 2020·Molecular and Cellular Probes·Carolina Villamil VallejoJohn Emerson Leguizamon
Dec 19, 2020·Comprehensive Reviews in Food Science and Food Safety·Carolina Ripolles-AvilaJosé-Juan Rodríguez-Jerez
Mar 30, 2021·Avian Pathology : Journal of the W.V.P.A·Mms SaraivaA Berchieri
May 2, 2021·Journal of Food Science·Song GaoJufang Wang
May 29, 2021·Analytical and Bioanalytical Chemistry·Nadja E Solis-MarcanoCarlos R Cabrera
Jun 23, 2021·Applied Microbiology and Biotechnology·Xuanqiang GuGuoqiang Zhu
Jun 27, 2021·Archives of Microbiology·Jingxuan ShiJinhai Huang
Jul 24, 2021·Biotechnology and Applied Biochemistry·Sebnem SeherlerIlke Anac Sakir
Aug 5, 2021·Electrophoresis·Nur Nasyifa Mohd MaidinMohd Ambri Mohamed

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