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

Diagn Microbiol Infect Dis. 2016 Mar;84(3):236-9. doi: 10.1016/j.diagmicrobio.2015.11.002. Epub 2015 Nov 6.

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.

Keywords: Campylobacter coli; Campylobacter jejuni; Ciprofloxacin-resistance; Degenerated primers; Fluoroquinolones; MAMA PCR; gyrA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animal Diseases / epidemiology
  • Animal Diseases / microbiology
  • Animals
  • Anti-Bacterial Agents / pharmacology*
  • Bacterial Proteins / chemistry
  • Bacterial Proteins / genetics
  • Base Sequence
  • Campylobacter Infections / epidemiology
  • Campylobacter Infections / microbiology
  • Campylobacter coli / drug effects*
  • Campylobacter coli / genetics*
  • Campylobacter jejuni / drug effects*
  • Campylobacter jejuni / genetics*
  • DNA Gyrase / chemistry
  • DNA Gyrase / genetics
  • Drug Resistance, Bacterial*
  • Genotype
  • Humans
  • Molecular Sequence Data
  • Phenotype
  • Polymerase Chain Reaction* / methods
  • Polymorphism, Genetic
  • Prevalence
  • Public Health Surveillance
  • Quinolones / pharmacology*
  • Sequence Alignment
  • Spain / epidemiology

Substances

  • Anti-Bacterial Agents
  • Bacterial Proteins
  • Quinolones
  • DNA Gyrase