A capillary polymerase chain reaction for Salmonella detection from poultry meat

Abstract

Aims: In this study, a capillary polymerase chain reaction (cPCR) was applied for Salmonella detection from poultry meat. Methods and Results: Salmonella detection limits of the optimized cPCR were determined with DNA templates from the samples of tetrathionate broth (TTB), Rappaport Vassiliadis broth (RVB) and selenite cystine broth (SCB) artificially contaminated with 10‐fold dilutions of 6 × 108 CFU ml−1 of pure Salmonella enterica ssp. enterica serovar Enteritidis 64K stock culture. Detection limits of cPCR from TTB, RVB and SCB were found as 6, 6 × 101 and 6 × 104 CFU ml−1, respectively. In addition, detection limits of bacteriology were also determined as 6 CFU ml−1 with TTB and SCB, and 6 × 101 CFU ml−1 with RVB. A total of 200 samples, consisting of 100 chicken and 100 turkey meat samples, were tested with optimized cPCR and bacteriology. Eight and six per cent of the chicken meat samples were found to harbour Salmonella by cPCR and standard bacteriology, respectively. Of six Salmonella isolates, four belonged to serogroup D, two to serogroup B. Conclusions: The TTB cultures of both artificially and naturally contaminated samples were found to be superior to those of RVB and SCB cultures in their cPCR results. This cPCR, utilizing template from 18‐h TTB primary enrichment broth culture, takes approximately 40 min in the successful detection of Salmonella from poultry meat. Significance and Impact of the Study: This study shows that cPCR from TTB enrichment culture of poultry meat would enable rapid detection of Salmonella in laboratories with low sample throughput and limited budget.

A comparative study on the use of real time polymerase chain reaction (RT-PCR) and standard isolation techniques for the detection of Salmonellae in broiler chicks

“…In addition, human gastroenteritis as a result of infection with poultry-associated Salmonellae is a well-known food-borne zoonosis and of health burden [16]. Previous researchers have shown that conventional methods should be carried out along with molecular techniques using RT-PCR [17]. Accordingly, this study has been done to compare conventional cultural isolation methods and RT-PCR technique for detection of Salmonellae in broiler chicks.…”

Rapid and accurate detection of Escherichia coli growth by fluorescent pH-sensitive organic nanoparticles for high-throughput screening applications

“…Although PCR and its variants can detect bacteria within a relatively short time (Gopinath et al, 2014;Noble and Weisberg, 2005), issues like primer design and PCRinhibitory effect of complex food matrices, in addition to its high cost, make PCR detection difficult to be a routine procedure (Gunaydin et al, 2007). Another effective method is enzyme-linked immunoabsorbent assay (ELISA).…”

Monitoring of Pollution of Salmonella sp. in Raw Milk Using Virulence Gen Marker

“…Molecular testing was carried out by detecting the presence of virulence genes using S139-S141 specific primers with nucleotide sequences based on inva gene markers 5'GTG AAA TTA TCG CCA CGT TCG GGC AA-3 'and 5' TCA TCG CAC CGT CAA AGG AAC C-3 '. The condition of PCR uses 35 cycles of amplification with the pre-denaturation stage using a temperature of 95°C for 5 minutes, denaturation at 94°C for 30 seconds, annealing at 60°C for 1 minute, extension at 72°C for 30 seconds, and final extension at 72°C for 10 minutes (De clercq, 2007;Rahn et al, 1992;Gunaydin, 2007;Malomy, et al, 2003).…”