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Detection and quantification of Campylobacter jejuni and Campylobacter coli using real‐time multiplex PCR
Author(s) -
Toplak N.,
Kovač M.,
Piskernik S.,
Možina S. Smole,
Jeršek B.
Publication year - 2012
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/j.1365-2672.2012.05235.x
Subject(s) - campylobacter jejuni , library science , campylobacter , clinical microbiology , campylobacter coli , multiplex , medicine , microbiology and biotechnology , biology , computer science , bioinformatics , bacteria , genetics
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.