1887

Abstract

This study describes a multiplex PCR assay for the detection of antibiotic resistance genes and the SXT element in . Conditions were optimized to amplify fragments of (encoding sulfamethoxazole resistance), (O1-specific trimethoprim resistance), (O139-specific trimethoprim resistance), (streptomycin B resistance) and the SXT element simultaneously in one PCR. This multiplex PCR was evaluated on 142 isolates and the results correlated with the phenotypic antibiotic data obtained using a disc diffusion assay and a colony blot assay. Thus this one-step PCR can be used as a simple, rapid and accurate method for identification of antibiotic resistance profiles and could be used for the surveillance of the spread of antibiotic resistance determinants in epidemiological and environmental studies.

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2007-03-01
2024-12-06
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References

  1. Amita Choudhury., R S., Thungapathra M., Ramamurthy T., Nair G. B., Ghosh A. 2003; Class 1 integrons and SXT elements in El Tor strains isolated before, and after 1992 Vibrio cholerae outbreak, Calcutta, India. Emerg Infect Dis 9:500–502 [CrossRef]
    [Google Scholar]
  2. Bauer A. W., Kirby W. M. M., Sherris J. C., Truck M. 1966; Antibiotic susceptibility testing by a standard single diffusion method. Am J Clin Pathol 45:493–496
    [Google Scholar]
  3. Bhanumathi R., Sabeena F., Isac S. R., Shukla B. N., Singh D. V. 2003; Molecular characterization of Vibrio cholerae O139 Bengal isolated from water and the aquatic plant Eichhornia crassipes in the River Ganga, Varanasi, India. Appl Environ Microbiol 69:2389–2394 [CrossRef]
    [Google Scholar]
  4. Dalsgaard A., Forslund A., Petersen A., Brown D. J., Dias F., Monteiro S., Molbak K., Aaby P., Rodrigues A., Sandström A. 2000; Class 1 integron-borne, multiple-antibiotic resistance encoded by a 150-kilobase conjugative plasmid in epidemic Vibrio cholerae O1 strains isolated in Guinea-Bissau. J Clin Microbiol 38:3774–3779
    [Google Scholar]
  5. Dalsgaard A., Forslund A., Sandvang D., Arntzen L., Keddy K. 2001; Vibrio cholerae O1 outbreak isolates in Mozambique and South Africa in 1998 are multiple-drug resistant, contain the SXT element and the aadA2 gene located on class 1 integrons. J Antimicrob Chemother 48:827–838 [CrossRef]
    [Google Scholar]
  6. Falbo V., Carattoli A., Tosini F., Pezzella C., Dionisi A. M., Luzzi I. 1999; Antibiotic resistance conferred by a conjugative plasmid and a class 1 integron in Vibrio cholerae O1 El Tor strains isolated in Albania and Italy. Antimicrob Agents Chemother 43:693–696
    [Google Scholar]
  7. Feinberg A., Vogelstein B. 1984; A technique for rapid labeling of DNA restriction endonuclease fragments to high specific activity. Anal Biochem 137:266–267 [CrossRef]
    [Google Scholar]
  8. Glass R. I., Huq M. I., Lee J. V., Threlfall E. J., Khan M. R., Alim A. R., Rowe B., Gross R. J. 1983; Plasmid-borne multiple drug resistance in Vibrio cholerae serogroup O1, biotype El Tor: evidence for a point-source outbreak in Bangladesh. J Infect Dis 147:204–209 [CrossRef]
    [Google Scholar]
  9. Hochhut B., Waldor M. K. 1999; Site specific integration of the non-conjugal Vibrio cholerae SXT element into prfC . Mol Microbiol 32:99–110 [CrossRef]
    [Google Scholar]
  10. Hochhut B., Marrero J., Waldor M. K. 2000; Mobilization of plasmids and chromosomal DNA mediated by the SXT element, a constin found in Vibrio cholerae O139. J Bacteriol 182:2043–2047 [CrossRef]
    [Google Scholar]
  11. Hochhut B., Lotfi Y., Mazel D., Faruque S. M., Woodgate R., Waldor M. K. 2001; Molecular analysis of antibiotic resistance gene clusters in Vibrio cholerae O139 and O1 SXT constins. Antimicrob Agents Chemother 45:2991–3000 [CrossRef]
    [Google Scholar]
  12. Iwanaga M., Toma C., Miyazato T., Insisiengmay S., Nakasone N., Ehara M. 2004; Antibiotic resistance conferred by a class 1 integron and SXT constin in Vibrio cholerae O1 strains isolated in Laos. Antimicrob Agents Chemother 48:2364–2369 [CrossRef]
    [Google Scholar]
  13. Keasler S. P., Hall R. H. 1993; Detecting and biotyping V. cholerae O1 with multiplex polymerase chain reaction. Lancet 341:1661 [CrossRef]
    [Google Scholar]
  14. Lindenbaum J., Greenough W. B., Islam M. R. 1967; Antibiotic therapy of cholera. Bull World Health Organ 36:871–883
    [Google Scholar]
  15. Mitra R., Basu A., Datta D., Nair G. B., Takeda Y. 1996; Resurgence of Vibrio cholerae O139 Bengal with altered antibiogram in Calcutta, India. Lancet 348:1181
    [Google Scholar]
  16. Ramamurthy T., Garg S., Sharma R., Bhattacharya S. K., Nair G. B., Shimada T., Takeda T., Karasawa T., Kurazano H. other authors 1993; Emergence of a novel strain of Vibrio cholerae with epidemic potential in southern and eastern India. Lancet 341:703–704
    [Google Scholar]
  17. Sambrook J., Russell D. W. 2001 Molecular Cloning: a Laboratory Manual , 3rd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  18. Sinha S., Chakraborty R., De K., Khan A., Datta S., Ramamurthy T., Bhattacharya S. K., Takeda Y., Nair G. B. 2002; Escalating association of Vibrio cholerae O139 with cholera outbreaks in India. J Clin Microbiol 40:2635–2637 [CrossRef]
    [Google Scholar]
  19. Tabtieng R., Wattanasri S., Echeverria P., Seriwatana J., Bodhidatta L., Chatkaeomorakot A., Rowe B. 1989; An epidemic of Vibrio cholerae El Tor Inaba resistant to several antibiotics with a conjugative group C plasmid coding for type II dihydrofolate reductase in Thailand. Am J Trop Med Hyg 41:680–686
    [Google Scholar]
  20. Thungapathra M., Amita Sinha., K K, Choudhury S. R., Garg P., Ramamurthy T., Nair G. B., Ghosh A. 2002; Occurrence of antibiotic resistance gene cassettes aac ( 6 ′)- Ib , dfrA5 , dfrA12 , and ereA2 in class 1 integrons in non-O1, non-O139 Vibrio cholerae strains in India. Antimicrob Agents Chemother 46:2948–2955 [CrossRef]
    [Google Scholar]
  21. Waldor M. K., Tschape H., Mekalanos J. J. 1996; A new type of conjugative transposon encodes resistance to sulfamethoxazole, trimethoprim, and streptomycin in Vibrio cholerae O139. J Bacteriol 178:4157–4165
    [Google Scholar]
  22. World Health Organization 1987; Program for control of diarrhoeal diseases. In Manual for Laboratory Investigation of Acute Infection , CDD/83.3 Geneva: World Health Organization;
    [Google Scholar]
  23. World Health Organization 1993 Guidelines for Cholera Control Geneva: World Health Organization;
    [Google Scholar]
  24. Yam W.-C., Yuen K.-Y., Wong S.-S.-Y., Que T.-L. 1994; Vibrio cholerae O139 susceptible to vibriostatic agent O/129 and co-trimoxazole. Lancet 344:404–405 [CrossRef]
    [Google Scholar]
  25. Yamamoto T., Nair G. B., Albert M. J., Parodi C., Takeda Y. 1994; In vitro susceptibility of Vibrio cholerae O1 and O139 to antimicrobial agents and appearance of drug resistance plasmids in Vibrio cholerae . In Proceedings of the 30th US–Japan Joint Conference on Cholera pp. 52–57 US–Japan Cooperative Medical Science Program for Cholera and Related Diarrheal Diseases Panel; Fukuara, Japan:
    [Google Scholar]
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