Characterization of non-classical quinolone resistance in Salmonella enterica serovar Typhi: Report of a novel mutation in gyrB gene and diagnostic challenges

Abstract

Objective

To establish the relative importance of Salmonella enterica serovar Typhi with non-classical quinolone resistance.

Methods

Eight hundred and ninety-one isolates of S. Typhi, isolated between 2004 and 2011, were tested for antibiotic susceptibility determination using disc diffusion and E-test. The mechanisms of fluoroquinolone resistance were studied in a sub-set of the NAL^S (nalidixic acid susceptible) isolates by wave nucleic acid fragment analysis of PCR products from gyrA, gyrB, parC and parE and from the plasmid borne determinants: qnrA,B,S; aac(6′)-Ib-cr and qepA. To assess genetic relatedness multi-locus variable number tandem repeat analysis was carried out using five loci.

Results

Eighty isolates with a nalidixic acid MIC of <32 mg/L (NAL^S) and a ciprofloxacin MIC of >0.064 mg/L CIP^I (ciprofloxacin reduced susceptibility) were found. In 36 NAL^S CIP^I isolates two distinct genotypes were identified when compared with 16 susceptible controls: Group B (n = 34), mutation in gyrB at codon 464, NAL MIC of 3–12 mg/L and CIP MIC of 0.064–0.5 mg/L.; and Group C, mutation in gyrA at codon 83 (n = 2) NAL MIC of 16 mg/L and CIP MIC of 0.25–0.38 mg/L. Group B isolates were found in different strain backgrounds as defined by MLVA.

Conclusion

The use of nalidixic acid to screen for reduced susceptibility to fluoroquinolones in S. Typhi misses CIP^I-NAL^S isolates, an established phenotype in India.