Bita Bakhshi
Tarbiat Modares University, Iran
Title: A rapid and reliable single step multiplex PCR assay for detection and differentiation of toxigenic Campylobacter jejuni and Campylobacter coli
Biography
Biography: Bita Bakhshi
Abstract
Background: Campylobacteriosis is a zoonotic infectious disease caused by C. jejuni and C. coli. cadF gene is considered as a genus-specific gene while other genes are mainly used for discrimination at species level.
Objectives: This study aimed to develop a duplex PCR assay for simultaneous detection of toxigenic C. coli and C. jejuni, the two commonly encountered campylobacter species.
Materials & Methods: A multiplex PCR optimized with specific primers for detection and differentiation of both species as well as Cytolethal Distending Toxin (CDT) gene was used. To evaluate specificity and sensitivity of test a panel of different Campylobacter spp., they were tested together with several other intestinal bacterial pathogens. The limit of detection of reaction determined using serial dilutions of standard genomes.
Results: The multiplex PCR designed in this study showed that it can simultaneously detect and differentiate both toxigenic and non-toxigenic C. jejuni and C. coli with different product sizes. This assay with 100% specificity and sensitivity has a Limit of Detection (LOD) of about 14 and 0.7 µg/ml for C. jejuni and C. coli, respectively.
Conclusions: Newly designed single-step multiplex-PCR assay could successfully detect and differentiate between toxigenic and non-toxigenic C. jejuni and C. coli with high specificity and sensitivity which can be used as a quick, reliable and powerful diagnostic tool for detection of campylobacteriosis in clinical settings.