AIMS: We describe a real-time quantitative multiplex polymerase chain reaction (qmPCR) assay to identify and discriminate between isolates of Campylobacter jejuni and Campylobacter coli. METHODS AND RESULTS: Two novel sets of primers and hydrolysis probes were designed to amplify the unique DNA sequences within the hipO, ccoN and cadF genes that are specific to Camp. jejuni and Camp. coli. Using the designed optimized qmPCR assay conditions, the amplification efficiency is in range from 108 to 116%. These qmPCR assays are highly specific for Camp. jejuni and Camp. coli, as seen through testing of 40 Campylobacter strains and 17 non-Campylobacter strains. In chicken juice and tap water models spiked with known quantities of Camp. jejuni, qmPCR detected 10(2) -10(3) CFU ml(-1) within 4 h. CONCLUSIONS: The qmPCR assays developed in this study provide reliable and simultaneous detection and quantification of Camp. jejuni and Camp. coli, with good amplification reaction parameters. SIGNIFICANCE AND IMPACT OF THE STUDY: Following further validation, the qmPCR assay reported here has the potential to be applied to various sample types as an alternative and rapid methodology.
AIMS: We describe a real-time quantitative multiplex polymerase chain reaction (qmPCR) assay to identify and discriminate between isolates of Campylobacter jejuni and Campylobacter coli. METHODS AND RESULTS: Two novel sets of primers and hydrolysis probes were designed to amplify the unique DNA sequences within the hipO, ccoN and cadF genes that are specific to Camp. jejuni and Camp. coli. Using the designed optimized qmPCR assay conditions, the amplification efficiency is in range from 108 to 116%. These qmPCR assays are highly specific for Camp. jejuni and Camp. coli, as seen through testing of 40 Campylobacter strains and 17 non-Campylobacter strains. In chicken juice and tap water models spiked with known quantities of Camp. jejuni, qmPCR detected 10(2) -10(3) CFU ml(-1) within 4 h. CONCLUSIONS: The qmPCR assays developed in this study provide reliable and simultaneous detection and quantification of Camp. jejuni and Camp. coli, with good amplification reaction parameters. SIGNIFICANCE AND IMPACT OF THE STUDY: Following further validation, the qmPCR assay reported here has the potential to be applied to various sample types as an alternative and rapid methodology.
Authors: Elizabeth Minogue; Kate Reddington; Siobhan Dorai-Raj; Nina Tuite; Eoin Clancy; Thomas Barry Journal: J Ind Microbiol Biotechnol Date: 2013-06-20 Impact factor: 3.346
Authors: G Di Martino; A Piccirillo; M Giacomelli; D Comin; A Gallina; K Capello; F Buniolo; C Montesissa; L Bonfanti Journal: Poult Sci Date: 2018-08-01 Impact factor: 3.352
Authors: Steven C Ricke; Kristina M Feye; W Evan Chaney; Zhaohao Shi; Hilary Pavlidis; Yichao Yang Journal: Front Microbiol Date: 2019-01-23 Impact factor: 5.640
Authors: Akram Sarabi Asiabar; Hamid Asadzadeh Aghdaei; Samin Zamani; Saied Bokaie; Mohammad Reza Zali; Mohammad Mehdi Feizabadi Journal: Med J Islam Repub Iran Date: 2019-07-29