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Journal of Clinical Microbiology, May 2000, p. 1767-1771, Vol. 38, No. 5
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Identification of Enterotoxigenic Escherichia coli Harboring Longus Type IV Pilus Gene by DNA Amplification

Zita Gutiérrez-Cázarez,1 Firdausi Qadri,2 M. John Albert,2 and Jorge A. Girón1,*

Centro de Investigaciones en Ciencias Microbiológicas, Instituto de Ciencias, Benemérita Universidad Autónoma de Puebla, Puebla, México,1 and International Centre for Diarrhoeal Disease Research, Bangladesh (ICDDR,B), Dhaka-1000, Bangladesh2

Received 9 November 1999/Returned for modification 20 December 1999/Accepted 11 February 2000

DNA amplification of lngA, the structural gene of longus type IV pilus produced by human enterotoxigenic Escherichia coli (ETEC) was achieved by the use of specific oligonucleotide primers designed from the nucleotide sequence of lngA. A 630-bp fragment representing the entire lngA gene was amplified in eight prototype strains previously characterized as longus positive. Five ETEC strains producing colonization factor antigen III (CFA III) (also a type IV pilus) were also positive by PCR, confirming the DNA homology between CFA III and longus. None of the non-ETEC and non-E. coli enteropathogens studied showed the 0.63-kbp amplicon. The procedure thus detected only ETEC strains harboring type IV pili genes with or without other colonization factors. Except for five lngA PCR-positive, probe-positive strains, all lngA PCR-positive strains produced the pilin as demonstrated by immunoblotting. To test the amplification procedure in a clinical setting, a collection of 264 fresh clinical E. coli strains isolated from 88 Mexican children with diarrhea was screened by PCR. Among 82 ETEC isolates found, 30 (36.5%) were lngA PCR-positive. Twenty-seven percent of the children shed ETEC that possessed lngA. In parallel with DNA probes or PCR protocols to detect enterotoxin genes, the lngA PCR method may prove useful for detection of ETEC harboring type IV pilus genes in epidemiological studies.

* Corresponding author. Mailing address: Centro de Investigaciones en Ciencias Microbiológicas, Instituto de Ciencias, Benemérita Universidad Autónoma de Puebla, Edificio 76, 3r Piso, 14 Sur Ave., San Claudio, Ciudad Universitaria, C. P. 72570, Puebla, Puebla, México. Phone: (52 22) 33 20 10. Fax: (52 22) 44 45 18. E-mail: jgiron{at}siu.buap.mx.

Journal of Clinical Microbiology, May 2000, p. 1767-1771, Vol. 38, No. 5
0095-1137/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


This article has been cited by other articles:

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