Novel < i > rpsK < /i > / < i > rpsD < /i > primer-probe assay improves detection of < i > Campylobacter jejuni < /i > and < i > Campylobacter coli < /i > in human stool

by Francesca Schiaffino, Craig T. Parker, Paul F. Garcia Bardales, Steven Huynh, Katia Manzanares Villanueva, Evangelos Mourkas, Ben Pascoe, Pablo Pe ñataro Yori, Maribel Paredes Olortegui, Eric R. Houpt, Jie Liu, Kerry K. Cooper, Margaret N. KosekCampylobacter causes bacterial enteritis, dysentery, and growth faltering in children in low- and middle-income countries (LMICs).Campylobacter spp. are fastidious organisms, and their detection often relies on culture independent diagnostic technologies, especially in LMICs.Campylobacter jejuni andCampylobacter coli are most often the infectious agents and in high income settings together account for 95% ofCampylobacter infections. Several otherCampylobacter species have been detected in LMIC children at an increased prevalence relative to high income settings. After doing extensive whole genome sequencing of isolates ofC.jejuni andC.coli in Peru, we observed heterogeneity in the binding sites for the main species-specific PCR assay(cadF) and designed an alternativerpsKD-based qPCR assay to detect bothC.jejuni andC.coli. TherpsKD-based qPCR assay identified 23% moreC.jejuni/ C.coli samples than thecadF assay among 47Campylobacter genus positivecadF negative samples verified to haveC.jejuni and orC.coli with shotgun metagenomics. This assay can be expected to be useful in diagnostic studies of enteric infectious diseases and be useful in revising the attribution estimates ofCampylobacter in LMICs.
Source: PLoS Neglected Tropical Diseases - Category: Tropical Medicine Authors: Source Type: research