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Antimicrobial Agents and Chemotherapy, September 2003, p. 2732-2739, Vol. 47, No. 9
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.9.2732-2739.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Molecular Analysis of incHI1 Antimicrobial Resistance Plasmids from Salmonella Serovar Typhi Strains Associated with Typhoid Fever

John Wain,1,2,3* L. T. Diem Nga,1 Claire Kidgell,2 Keith James,4 Sarah Fortune,1,{dagger} To Song Diep,5 Tahir Ali,2,{ddagger} Peadar Ó Gaora,2 Christopher Parry,1,3 Julian Parkhill,4 Jeremy Farrar,1,3 Nicholas J. White,1,3 and Gordon Dougan2

Wellcome Trust Clinical Research Unit,1 Centre for Tropical Diseases, Ho Chi Minh City, Vietnam,5 Centre for Molecular Microbiology and Infection, Department of Biological Sciences and Division of Investigative Sciences, Imperial College, London,2 Sanger Institute, Wellcome Genome Campus, Hinxton, Cambridge,4 Centre for Tropical Medicine, Nuffield Department of Clinical Medicine, John Radcliffe Hospital, Oxford University, Oxford, United Kingdom3

Received 16 December 2002/ Returned for modification 4 May 2003/ Accepted 4 June 2003

The first outbreak of multidrug-resistant (MDR) typhoid fever in Vietnam was in 1993, and by 1995 nearly 90% of cases were MDR. Plasmid HCM1, sequenced in full, is an incHI1 plasmid from Salmonella enterica serovar Typhi strain CT18, isolated in Vietnam in 1993. Restriction analysis shows that pHCM1 shares a restriction fragment length polymorphism (RFLP) pattern with plasmids isolated from the first outbreak and 10 of 17 MDR plasmids isolated from sporadic cases occurring at the same time in Vietnam. A core region of pHCM1 has significant DNA sequence similarity to plasmid R27, isolated in 1961 from S. enterica in the United Kingdom. There are five regions of DNA in pHCM1 which are not present in R27. Two of these are putative acquisition regions; the largest is 34.955 kbp in length and includes sequences of several antibiotic resistance genes and several insertion sequences. The borders of this region are defined by two identical IS10 left elements, associated with an inversion of DNA or with a truncated Tn10 element. The second, smaller region is 14.751 kbp and carries a trimethoprim resistance gene dfr14A cassette associated with a class 1 integrase. In 1993 to 1994, restriction analysis revealed some variations in the structures of Salmonella serovar Typhi MDR plasmids which were mapped to the two putative acquisition regions and three smaller variable regions. In 1996 a single RFLP type, RFLP7, was found to carry the dfrA7 and sul-1 genes, which were not present on R27 or pHCM1. This plasmid type appears to have a selective advantage over other plasmids with the same resistance phenotype.

* Corresponding author. Mailing address: Centre for Molecular Microbiology and Infection, 3rd Floor Flower's Building, Imperial College, Armstrong Rd., London SW7 2AZ, United Kingdom. Phone: 442075943199. Fax: 442075943069. E-mail: j.wain{at}ic.ac.uk.

{dagger} Present address: Division of Immunology and Infectious Diseases, Harvard School of Public Health, Boston, Mass.

{ddagger} Present address: Academic Pediatrics, St. Mary's Medical School, Imperial College, London, United Kingdom.

Antimicrobial Agents and Chemotherapy, September 2003, p. 2732-2739, Vol. 47, No. 9
0066-4804/03/$08.00+0     DOI: 10.1128/AAC.47.9.2732-2739.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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