Premium
Detection of Salmonella spp., Y ersinia enterocolitica , L isteria monocytogenes and C ampylobacter spp. by real‐time multiplex PCR using amplicon DNA melting analysis and probe‐based assay
Author(s) -
Skerniškytė Jūratė,
Armalytė Julija,
Kvietkauskaitė Raimonda,
Šeputienė Vaida,
Povilonis Justas,
Sužiedėlienė Edita
Publication year - 2016
Publication title -
international journal of food science and technology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.831
H-Index - 96
eISSN - 1365-2621
pISSN - 0950-5423
DOI - 10.1111/ijfs.12990
Subject(s) - yersinia enterocolitica , listeria monocytogenes , salmonella , campylobacter , amplicon , microbiology and biotechnology , biology , multiplex , yersinia , taqman , multiplex polymerase chain reaction , real time polymerase chain reaction , bacteria , gene , polymerase chain reaction , genetics
Summary 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 ± 0.5, 84 ± 0.5, 81.5 ± 0.5 and 90.5 ± 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 × 10 2 to 3.1 × 10 4 and 9.8 × 10 2 to 1.9 × 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.