African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12487

Full Length Research Paper

Multiplex polymerase chain reaction for detection and characterization of shiga toxigenic Escherichia coli (STEC)

Moussa, I. M.1*, Ashgan, M. H.2, Alwathnani, H. A.3, Mohamed, Kh. F.4 and Al-Doss, A. A.1
1Center of Excellence in Biotechnology, King Saud University, P. O. 2460 Riyadh, King Saudi Arabia, Saudi Arabia. 2College of Applied Studies and Community Services, King Saud University, Saudi Arabia. 3Department of Botany and Microbiology, College of Science, King Saud University, Saudi Arabia. 4Department of Microbiology, Faculty of Veterinary Medicine, Cairo University, Egypt.  
Email: [email protected], [email protected]

  •  Accepted: 21 June 2010
  •  Published: 31 July 2010

Abstract

Escherichia coli is ubiquitous in the cow's environment that is contaminated by feces, and it is also a frequent cause of bovine mastitis. Thus, the present study was targeted at the rapid detection and characterization of shiga toxigenic E. coli(STEC) in bovine fecal and milk samples. Twenty two strains of E. coli (39.29%) were isolated from 56 diarrheic calves, while only 5 strains (20.83%) were isolated from apparently normal contact calves. Moreover, 20 strains of E. coli (25%) were isolated from milk samples collected from 80 animals suffering from mastitis and subclinical mastitis. E. coli serovars yielded from bacteriologi­cal examination of milk samples were similar to that of fecal samples. Serogroup-specific multiplex polymerase chain reaction (PCR) assay could detect all the bacteriologically positive samples as well as 4 strains (7.98%), O157:H7 and 3 strains (5.36%), O111 from diarrheic calves and 2 strains (8.33%), O111 from normal calves. Such samples were proved to be negative by bacteriological examination. Multiplex PCR for detection of genes encoding accessory STEC virulence factors, such as shiga toxin type-2 (stx2) and intimin gene (eaeA) revealed the specificity of such gene to O157:H7 serovars and small number of other sero-groups.

 

Key words: Escherichia coli, shiga toxigenic, diarrheic calves, multiplex PCR, intimin gene, O157:H7 serovars.

Abbreviation

STEC, Shiga toxigenic Escherichia colistx2, shiga toxin type-2;PCR, polymerase chain reaction; HC, hemorrhagic colitis; HUS, haemolytic uraemic syndrome; TSB, tryptic soy broth; DDW, deionized distilled water.