Malaysian Applied Biology Journal

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26-1-04

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Malays. Appl. Biol. (1997) 26(1): 19-23

DETECTION OF ENTEROTOXIGENIC ESCHERICHIA COLI ASSOCIATED WITH DIARRHOEA BY BIOTINYLATED HEAT-STABLE (ST) DNA PROBE

SON RADU*1, RUSUL, G.2, KARIM, M. I. A.1 and YUSOFF, K.3

1 Department of Biotechnology, 2 Department of Food Science, 3 Department of Biochemistry and Microbiology, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia

ABSTRACT

Several studies have demonstrated the usefulness of recombinant DNA probes for identifying enterotoxigenic Escherichia coli (ETEC). In this study, an enzyme-labelled oligonucleotide probe for an ETEC heat-stable toxin (ST) gene was compared with the standard suckling mouse assay for their ability to identify ETEC in a group of adults experiencing acute episode of diarrhoea. The ST probe hybridized with 48% (10 of 21) of the E. coli colonies tested, whereas 38% (8 of 21) were positive by the suckling mouse bioassay. For the patient samples tested, three ST-probe positive were negative for suckling mouse bioassay. Plasmid DNA among the 10 ST-probe positive E. coli strains ranging in size between 1.6 kb to 35 kb were shown to hybridize with the biotinylated ST DNA probe. These results indicate that the biotinylated enterotoxin probe is useful in the diagnosis of enterotoxigenic E. coli strains by colony hybridization, and that ETEC play a modest role in the aetiology of diarrhoea among the group of patients examined.


ABSTRAK

Beberapa kajian telah menunjukkan penggunaan prob DNA untuk mengenalpasti Escherichia coli enterotoksigenik (ETEC). Dalam kajian ini, prob oligonukleotid yang dilabel dengan enzim untuk gen toksin tahan haba ETEC (ST) dibandingkan dengan 'suckling mouse assay' bagi melihat keberkesanan mereka mengesan ETEC dalam sekumpulan orang dewasa yang mengalami cirit birit. Prob ST berhibrid dengan 48% (10 dari 21) koloni E. coli yang diuji, manakala 38% (8 dari 21) adalah positif dengan cara 'suckling mouse assay'. Dari sampel yang diuji, tiga yang prob-positif adalah negatif dengan 'suckling mouse bioassay'. Plasmid DNA dikalangan 10 asingan E. coli yang prob-positif bersaiz antara 1.6 kb dan 35 kb menghibrid kepada prob ST DNA yang dilabelkan dengan biotin. Keputusan yang diperolehi menunjukkan bahawa prob snterotoksin yang dilabelkan dengan biotin amat berguna dalam pengesanan asingan E. coli dengan cara hibridisasi koloni, dan juga ETEC memainkan peranan dalam kejadian cirit birit dikalangan pesakit yang diuji.

Key words: Biotinylated DNA probe, enterotoxigenic E. coli, heat-stable enterotoxin, suckling mouse assay

REFERRENCES

Birnboim, H. C. and Doly, J. 1979. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Research, 7: 1513-1523.

Moseley, S. L., Echeverria, P., Seriwatana, J., Tirapat, C., Chaicumpa, W., Sakuldaipeara, T. and Falkow, S. 1982. Identification of enterotoxigenic Escherichia coli by colony hybridization using three enterotoxin gene probes. Journal of Infectious Disease, 145: 863-869.

Murray, B. E., Mathewson, J. J., DuPont, H. L. and Hill, W. E. 1987. Utility of oligodeoxyribo-nucleotide probes for detecting enterotoxigenic Escherichia coli. Journal of Infectious Disease, 155: 809-811.

Sack, D. A. and Sack, R. B. 1975. Test for enterotoxigenic Escherichia coli using Y-l adrenal cells in miniculture. Infection and Immunity, 11: 334-336.

Smith, N. W. and Gyles, C. L. 1970. The relationship between two apparently different enterotoxins produced by enteropathogenic Escherichia coli strains of porcine origin.

Southern, E. M. 1975. Detection of specific sequences among DNA fragments by gel electrophoresis. Journal of Molecular Biology, 98: 503-517.

Taylor, D. N. and Echeverria, P. 1986. Etiology and epidemiology of traveller's diarrhea in Asia. Review of Infectious Disease, 8 Suppl. 2: SI 36-S141.

Thompson, M. R., Brandwein, H., LaBine-Racke, M and Gianella, R. A. 1984. Simple and reliable enzyme-linked immunosorbant assay with monoclonal antibodies for detection ol Escherichia coli heat-stable enterotoxins. Journal of Clinical Microbiology, 20: 59-64.

Yolken, R. H., Greenberg, H. B., Merson, M. H. Sack, R. B. and Kapikian, A. Z. 1977. Enzyme-linked immunosorbant assay for detection ol Escherichia coli heat-labile enterotoxin. Journal of Clinical Microbiology, 6: 439-444.
 

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