Identification of the main quinolone resistance determinant in Campylobacter jejuni and Campylobacter coli by MAMA-DEG PCR

Among zoonotic diseases, campylobacteriosis stands out as the major bacterial infection producing human gastroenteritis. Antimicrobial therapy, only recommended in critical cases, is challenged by resistance mechanisms that should be unambiguously detected for achievement of effective treatments. Quinolone (ciprofloxacin) resistance of Campylobacter jejuni and Campylobacter coli, the2main Campylobacter detected in humans, is conferred by the mutation gyrA C-257-T, which can be genotyped by several methods that require a previous identification of the pathogen species to circumvent the sequence polymorphism of the gene. A multiplex PCR, based on degenerated oligonucleotides, has been designed for unambiguous identification of the quinolone resistance determinant in Campylobacter spp. isolates. The method was verified with 249 Campylobacter strains isolated from humans (141 isolates) and from the 3 most important animal sources for this zoonosis: poultry (34 isolates), swine (38 isolates), and cattle (36 isolates). High resistance to ciprofloxacin, MIC above 4 μg/mL, linked to the mutated genotype predicted by MAMA-DEG PCR (mismatch amplification mutation assay PCR with degenerated primers) was found frequently among isolates from the different hosts.

Keywords

MAMA PCR Degenerated primers gyrA Fluoroquinolones Ciprofloxacin-resistance Campylobacter jejuni Campylobacter coli

URI

http://hdl.handle.net/10400.8/13640

Citation

Lorena Hormeño, Gonzalo Palomo, María Ugarte-Ruiz, M. Concepción Porrero, Carmen Borge, Santiago Vadillo, Segundo Píriz, Lucas Domínguez, Maria J. Campos, Alberto Quesada, Identification of the main quinolone resistance determinant in Campylobacter jejuni and Campylobacter coli by MAMA-DEG PCR, Diagnostic Microbiology and Infectious Disease, Volume 84, Issue 3, 2016, Pages 236-239, ISSN 0732-8893, https://doi.org/10.1016/j.diagmicrobio.2015.11.002