TY - JOUR

T1 - Reverse transcriptase real-time PCR for detection and quantification of viable Campylobacter jejuni directly from poultry faecal samples

A1 - Bui,Thanh Xuan

A1 - Wolff,Anders

A1 - Madsen,Mogens

A1 - Bang,Dang Duong

AU - Bui,Thanh Xuan

AU - Wolff,Anders

AU - Madsen,Mogens

AU - Bang,Dang Duong

PB - Elsevier France Editions Scientifiques et Medicales

PY - 2012

Y1 - 2012

N2 - Campylobacter spp. is the most common cause of bacterial diarrhoea in humans worldwide. Therefore, rapid and reliable methods fordetection and quantification of this pathogen are required. In this study, we have developed a reverse transcription quantitative real-time PCR(RT-qPCR) for detection and quantification of viable Campylobacter jejuni directly from chicken faecal samples. The results of this method anda DNA-based quantitative real-time PCR (qPCR) method were compared with those of a bacterial culture method. Using bacterial culture andRT-qPCR methods, viable C. jejuni cells could be detected for up to 5 days in both the C. jejuni spiked and the naturally contaminated faecalsamples. We found that no RT-qPCR signals were obtained when viable C. jejuni cells could not be counted by the culture method. In contrast,using a DNA-based qPCR method, dead or non-viable Campylobacter cells were detected, and all tested samples were positive, even after 20days of storage. The developed method for detection and quantification of viable C. jejuni cells directly from chicken faecal samples can be usedfor further research on the survival of Campylobacter in the environment.

AB - Campylobacter spp. is the most common cause of bacterial diarrhoea in humans worldwide. Therefore, rapid and reliable methods fordetection and quantification of this pathogen are required. In this study, we have developed a reverse transcription quantitative real-time PCR(RT-qPCR) for detection and quantification of viable Campylobacter jejuni directly from chicken faecal samples. The results of this method anda DNA-based quantitative real-time PCR (qPCR) method were compared with those of a bacterial culture method. Using bacterial culture andRT-qPCR methods, viable C. jejuni cells could be detected for up to 5 days in both the C. jejuni spiked and the naturally contaminated faecalsamples. We found that no RT-qPCR signals were obtained when viable C. jejuni cells could not be counted by the culture method. In contrast,using a DNA-based qPCR method, dead or non-viable Campylobacter cells were detected, and all tested samples were positive, even after 20days of storage. The developed method for detection and quantification of viable C. jejuni cells directly from chicken faecal samples can be usedfor further research on the survival of Campylobacter in the environment.

KW - RT-qPCR

KW - Campylobacter jejuni

KW - Chicken faeces

KW - Campylobacter survival

KW - mRNA

U2 - 10.1016/j.resmic.2011.10.007

DO - 10.1016/j.resmic.2011.10.007

JO - Research in Microbiology

JF - Research in Microbiology

SN - 0923-2508

IS - 1

VL - 163

SP - 64

EP - 72

ER -