Detection of mutations in the gyrA gene and class I integron from quinolone-resistant Salmonella enterica serovar Choleraesuis isolates in Taiwan

Abstract

The quinolone resistance-determining regions (QRDRs) of the gyrA gene of quinolone-resistant Salmonella enterica serovar Choleraesuis isolates were sequenced. Four types of point mutation, Ser-83-to-Phe (TCC→TTC), Ser-83-to-Tyr (TCC→TAC), Asp-87-to-Gly (GAC→GGC), and Asp-87-to-Asn (GAC→AAC), were found. PCR-RFLP and MAS-touch down PCR were performed on fifty swine clinical isolates of S. enterica serovar Choleraesuis (Nal R) collected during 1997–2002. The analysis indicated seven isolates with point mutations in codon 83, 13 with point mutations in codon 87, and 30 with double mutations in both codons 83 and 87. The MICs of enrofloxacin of the isolates with a single mutation in codon 83 or 87 were <2 μg/ml, while the MICs of the isolates with double mutations in both codon 83 and 87 ranged from 2 to 64 μg/ml. A class I integron comprised of dhfr, orfF and aad2 was also identified in both human and swine S. enterica serovar Choleraesuis isolates. These results indicate that PCR-RFLP and MAS-touchdown PCR assays can be used for surveillance of gyrA gene mutations, which are important for fluoroquinolone resistance in Salmonella. Isolates with double mutations in gyrA codons 83 and 87 are the major type of quinolone-resistant Salmonella isolated from swine in Taiwan. A surveillance system may be applied to the swine industry to monitor the emergence of fluoroquinolone and/or multi-drug-resistant S. enterica serovar Choleraesuis in Taiwan.