title: Identification of the main quinolone resistance determinant in Campylobacter jejuni and Campylobacter coli by MAMA-DEG PCR.
abstract: 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, the 2 main 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 4μg/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.
authors.affiliation: Departamento de Sanidad Animal, Facultad de Veterinaria, Universidad de Extremadura, Cáceres, Spain.
authors.forename: Lorena
authors.initials: L
authors.lastname: Hormeño
authors.affiliation: Departamento de Sanidad Animal, Facultad de Veterinaria, Universidad de Extremadura, Cáceres, Spain.
authors.forename: Gonzalo
authors.initials: G
authors.lastname: Palomo
authors.affiliation: Centro de Vigilancia Sanitaria Veterinaria (VISAVET), Universidad Complutense Madrid, Madrid, Spain.
authors.forename: María
authors.initials: M
authors.lastname: Ugarte-Ruiz
authors.affiliation: Centro de Vigilancia Sanitaria Veterinaria (VISAVET), Universidad Complutense Madrid, Madrid, Spain.
authors.forename: M Concepción
authors.initials: MC
authors.lastname: Porrero
authors.affiliation: Departamento de Sanidad Animal, Facultad de Veterinaria, Universidad de Córdoba, Córdoba, Spain.
authors.forename: Carmen
authors.initials: C
authors.lastname: Borge
authors.affiliation: Departamento de Sanidad Animal, Facultad de Veterinaria, Universidad de Extremadura, Cáceres, Spain.
authors.forename: Santiago
authors.initials: S
authors.lastname: Vadillo
authors.affiliation: Departamento de Sanidad Animal, Facultad de Veterinaria, Universidad de Extremadura, Cáceres, Spain.
authors.forename: Segundo
authors.initials: S
authors.lastname: Píriz
authors.affiliation: Centro de Vigilancia Sanitaria Veterinaria (VISAVET), Universidad Complutense Madrid, Madrid, Spain.
authors.forename: Lucas
authors.initials: L
authors.lastname: Domínguez
authors.affiliation: MARE-Marine and Environmental Sciences Centre, ESTM, Instituto Politécnico de Leiria, 2520-641 Peniche, Portugal. Electronic address: mjgcampos@ipleiria.pt.
authors.forename: Maria J
authors.initials: MJ
authors.lastname: Campos
authors.affiliation: Departamento de Bioquímica, Biología Molecular y Genética, Facultad de Veterinaria, Universidad de Extremadura, Cáceres, Spain.
authors.forename: Alberto
authors.initials: A
authors.lastname: Quesada
mesh_terms: D000820:Animal Diseases / Q000453:epidemiology / Q000382:microbiology; D000818:Animals; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics; D001483:Base Sequence; D002169:Campylobacter Infections / Q000453:epidemiology / Q000382:microbiology; D017000:Campylobacter coli / Q000187:drug effects* / Q000235:genetics*; D016123:Campylobacter jejuni / Q000187:drug effects* / Q000235:genetics*; D027081:DNA Gyrase / Q000737:chemistry / Q000235:genetics; D024881:Drug Resistance, Bacterial; D005838:Genotype; D006801:Humans; D008969:Molecular Sequence Data; D010641:Phenotype; D016133:Polymerase Chain Reaction / Q000379:methods; D011110:Polymorphism, Genetic; D015995:Prevalence; D062486:Public Health Surveillance; D015363:Quinolones / Q000494:pharmacology*; D016415:Sequence Alignment; D013030:Spain / Q000453:epidemiology
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D015363:Quinolones; D027081:DNA Gyrase
keywords: Campylobacter coli; Campylobacter jejuni; Ciprofloxacin-resistance; Degenerated primers; Fluoroquinolones; MAMA PCR; gyrA