JAC Advance Access originally published online on June 9, 2006
Journal of Antimicrobial Chemotherapy 2006 58(2):288-296; doi:10.1093/jac/dkl228
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Molecular characteristics of class 1 and class 2 integrons and their relationships to antibiotic resistance in clinical isolates of Shigella sonnei and Shigella flexneri
1 Microbiology Laboratory, Hangzhou Center for Disease Control and Prevention Hangzhou 310006, People's Republic of China 2 Institute for Infectious Disease, Zhejiang University Hangzhou 310003, People's Republic of China
Received 9 February 2006; returned 21 March 2006; revised 29 April 2006; accepted 5 May 2006
*Correspondence address: Microbiology Laboratory, Hangzhou Center for Disease Control and Prevention, Hangzhou, 310006, People's Republic of China. Tel: +86-571-85177696; Fax: +86-571-85165007; E-mail: jingcaopan{at}sina.com
Objectives: To analyse the gene cassettes and determine the roles of class 1 and class 2 integrons in antibiotic-resistant strains of Shigella sonnei (n = 31) and Shigella flexneri (n = 33).
Methods: Various molecular techniques, including PCR and Southern-blotting analysis, were used to analyse various markers of class 1 and class 2 integrons in these 64 S. sonnei and S. flexneri isolates collected in Hangzhou, China. The gene cassette arrays in integrons were identified by DNA sequencing and/or restriction fragment length polymorphism. Two genomic DNA fragments, one containing intI1 from a S. flexneri isolate that contains intI1 but lacks 3'-conserved region and another containing intI2 from a S. sonnei isolate, were cloned into pUC19 vectors and sequenced. The links between integron gene cassette arrays and antibiotic resistance were analysed.
Results: Class 2 integrons were present in 80.6% (25/31) of the S. sonnei isolates and 87.9% (29/33) of the S. flexneri isolates. All of these integron 2-positive isolates contained constant gene cassette arrays of dfrA1 + sat1 + aadA1 which confer resistance to trimethoprim and streptomycin. It was demonstrated that the class 2 integron was located in the Tn7 region inside the attTn7 locus downstream of glmS in Shigella. Class 1 integrons were found in 9.4% (6/64) of Shigella spp. isolates. An atypical class 1 integron without a 3'-conserved segment on the Shigella chromosome, termed Shigella atypical class 1 integron (SAI), was present in 84.9% (28/33) of S. flexneri isolates. The SAI contained two gene cassettes, blaOXA30 and aadA1; however, the SAI conferred resistance to ampicillin, but not to streptomycin, in Escherichia coli host. The blaOXA30 and aadA1 cassettes of the SAI seemed to be always coordinately excised or integrated.
Conclusions: Multiple and complex mechanisms involving mobile genetic elements in class 1 and class 2 integrons and antibiotic resistance have been developed in the evolution of Shigella strains.
Keywords: Tn7 , blaOXA30 , dfrA1 , aadA1
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