Detection of Salmonella spp., Yersinia enterocolitica,Listeria monocytogenes and Campylobacter spp. by real‐time multiplex PCR using amplicon DNA melting analysis and probe‐based assay

Abstract

Syto9 and probe-based multiplex real-time PCR assays for simultaneous detection of a group of foodborne pathogens (named SYLC group), targeting Salmonella spp. (invA gene), Yersinia enterocolitica (ystA gene), Listeria monocytogenes (hly gene) and Campylobacter spp. (rrna gene), have been developed. The Syto9 assay generates amplicon DNA melting curve with four peaks of 86.5 AE 0.5, 84 AE 0.5, 81.5 AE 0.5 and 90.5 AE 0.5°C corresponding Salmonella spp., Y. enterocolitica, L. monocytogenes and Campylobacter spp. targets, respectively. The sensitivities of the Syto9 and TaqMan assays in artificially inoculated chicken wing rinses were in a range of 3.2 9 10 2 to 3.1 9 10 4 and 9.8 9 10 2 to 1.9 9 10 4 colony-forming units per millilitre, respectively, depending on the pathogen. All tested target strains (n = 100) were correctly detected by the both assays, whereas nontarget strains (n = 100) demonstrated no cross-reactivity representing 100% specificity. The assays are suitable for application in qualitative and quantitative detection of SYLC group pathogens in food matrices.

Reductionin Listeria monocytogenes, Salmonella enterica and Escherichia coli O157:H7 in vitro and on tomato by sophorolipid and sanitiser as affected by temperature and storage time

“…Among these, Escherichia coli O157:H7, Salmonella enterica serovars and Listeria monocytogenes have been the most frequently encountered bacterial pathogens in outbreaks on minimally processed produce and food (Phillips, 2016;Skerni skyt_ e et al, 2016). Among these, Escherichia coli O157:H7, Salmonella enterica serovars and Listeria monocytogenes have been the most frequently encountered bacterial pathogens in outbreaks on minimally processed produce and food (Phillips, 2016;Skerni skyt_ e et al, 2016).…”

“…Bacterial pathogens account for the vast majority of produce and food contamination. Among these, Escherichia coli O157:H7, Salmonella enterica serovars and Listeria monocytogenes have been the most frequently encountered bacterial pathogens in outbreaks on minimally processed produce and food (Phillips, 2016;Skerni skyt_ e et al, 2016). Therefore, inactivation of foodborne pathogens and their control to enhance the safety of produce and other foods is of immediate concern particularly on postharvest commodities (Olmez & Kretzschmar, 2009).…”

Detection of Listeria spp. and Listeria monocytogenes in biological samples by SYBR Green I and TaqMan probe-based real-time PCRs

“…( 2 ) and Skerniškyté et al . ( 23 ) achieved 100% specificity in TaqMan probe-based real-time PCR using hly gene. Our results also demonstrated specificity reaching 100% in all SYBR Green I and TaqMan probe-based real-time PCRs with the use 23S rDNA and hly A genes.…”

Rapid detection of Listeria monocytogenes using fluorescence immunochromatographic assay combined with immunomagnetic separation technique

“…In recent years, many new biotechnology‐based techniques have been studied to improve the LM detection efficiency. Molecular biology methods, such as nucleic acid probe and gene chip, have high sensitivity, but need expensive equipment and skilled operation (Feng et al ., ; Skerniškyt≐ et al ., ). Various types of immunological detection methods like enzyme‐linked immunosorbent assay [(ELISA) Mendonça et al ., ; Mizutani et al ., ;], colloidal gold immunochromatography (Shan et al ., ) and immunomagnetic separation (IMS) method (Uusitalo et al ., ) have widely been used in pathogens and toxins detection.…”