Detection of eae, bfpA, espA Genes on Diarrhoeagenic Strains of Escherichia coli Isolates

Agnes Sri Harti; Susi Iravati; Widya Asmara

doi:10.22146/ijbiotech.7563

Detection of eae, bfpA, espA Genes on Diarrhoeagenic Strains of Escherichia coli Isolates

https://doi.org/10.22146/ijbiotech.7563

Agnes Sri Harti^(1*), Susi Iravati⁽²⁾, Widya Asmara⁽³⁾

(1) Research Center for Biotechnology, Gadjah Mada University,Yogyakarta 55281, Indonesia.Faculty of Biology, Setia Budi University, Surakarta, Indonesia
(2) Department of Microbiology, Gadjah Mada University School of Medicine, Yogyakarta 55281,Indonesia
(3) Department of Microbiology, Faculty of Veterinary Medicine, Gadjah Mada University, Yogyakarta55281, Indonesia
(*) Corresponding Author

Abstract

The Enteropathogenic Escherichia coli (EPEC) is one of pathogenic strain of diarrheagenic E. coli group in children and infant that occurs in developing countries. The significant virulence factors in pathogenic EPEC are eaeA (E. coli attaching- effacing ), bfpA (bundle-forming pilus A) and espA (encoding secreted protein A) genes. The use of DNA probes to detect the virulence genes in E. coli in Indonesia is not common yet. In this experiment the gene fragments of eae, bfpA, and espA were used as probes to detect the EPEC among E. coli isolates from stool specimensin of diarrheic children attending Public Health Centers in Yogyakarta. The DNA samples were isolated from 49 diarrheagenic E. coli isolates. The DNA probes of eae, bfpA and espA were obtained by amplification of DNA fragment of EPEC O126 using PCR technique. Furthermore, those probes were used to identify the presence of those genes among E. coli isolates using hybridization technique. The results showed that 42 (85.7%) isolates were espA+, 25 isolates (51%) were eaeA+ (EPEC strains). Therefore among 25 isolates of EPEC, 20 isolates (80 %) among EPEC were bfpA+ (typical EPEC strains).

Keywords

DNA probe, eae, bfpA, espA, EPEC

Full Text:

PDF

References

Afset, J.E., Bergh, K., and Bevanger, L., 2003. H i g h p r e v a l e n c e o f a t y p i c a l enteropatogenic Escherichia coli (EPEC) in Norwegian children with diarrhoea. J. Med. Microbiol., 52, 1015–1019.

Aranda, K.R.S., Fagundes-Neto, U., and Scaletsky, I.C.A., 2004. Evaluation of multiplex PCRs for diagnosis of i n f e c t i o n w i t h d i a r r h e a g e n i c Escherichia coli and Shigella spp. J. Clin. Microbiol., 42 (12), 5849–5853.

Bekal, S., Brousseau, R., Masson, L., Prefontaine, G., Fairbrother, J., and Harel, J., 2003. Rapid identification of Escherichia coli pathotypes by virulence gene detection with DNA microarrays. J. Clin. Microbiol., 41 (5), 2113–2125.

Cleary, J., Lai, L-C., Shaw, R.K., Straatman- Iwanowska, A., Donneberg, M.S., Frankel, G., and Knutton, S., 2004. Enteropathogenic Escherichia coli (EPEC) adhesion to intestinal epitel cells: role of BFP, EspA filaments and Intimin. Microbiology, 150, 527–538.

Kuhne, S.A., Hawes, W.S., La Ragione, R.M., Woodward, M.J., Whitelam, G.C., and Gough, K.C., 2004. Isolation of recombinant antibodies against EspA and Intimin of Escherichia coli O157 : H7. J. Clin. Microbiol., 42 (7), 2966–2976.

Knutton, S., Rosenshine, I., Pallen, M.J., Nisan, I., Neves, B.C. Bain, C., Wolf, C., Dougan, G. and Frankel, G., 1998. A novel Esp A- associated surface organelle of enteropathogenic Escherichia coli involved in protein translocation into epithelial cells. EMBO J., 17, 2166–2176.

Madigan, M.T., Martinko, J.M. and Parker, J., 2002. Brock biology of microorganism. 9th ed. Prentice Hall International, Inc.

Nataro, J.P. and Kaper, J.B., 1998. Diarrheagenic Escherichia coli. Clin. Microbiol. Rev., 11 (1), 142–201.

Shaw, R.K., Danielli, S., Ebel, F., Frankel, G., and Knutton, S., 2001. EspA filament – mediated protein translocation into red blood cells. Cell. Microbiol., 3, 213- 222.

Trabulsi, L.R., Keller, R., and Gomes, T.A.T., 2 0 0 2 . T y p i c a l a n d a t y p i c a l enteropathogenic Escherichia coli. Synopsis. J. Emerg. Infect. Dis., 8 5, 224-227.

DOI: https://doi.org/10.22146/ijbiotech.7563