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

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.