J Vet Sci.  2005 Mar;6(1):7-19.

Isolation and identification of Escherichia coli O157:H7 using different detection methods and molecular determination by multiplex PCR and RAPD

• Kim JY
• Kim SH
• Kwon NH
• Bae WK
• Lim JY
• Koo HC
• Kim JM
• Noh KM
• Jung WK
• Park KT
• Park YH

Abstract

Escherichia coli O157:H7 is recognized as a significant food-borne pathogen, so rapid identification is important for food hygiene management and prompt epidemiological investigations. The limited prevalence data on Shiga toxin-producing E. coli (STEC) and E. coli O157:H7 in foods and animals in Korea made an assessment of the risks difficult, and the options for management and control unclear. The prevalence of the organisms was examined by newly developed kit-E. coli O157:H7 Rapid kit. For the isolation of E. coli O157:H7, conventional culture, immunomagnetic separation, and E. coli O157:H7 Rapid kit were applied, and multiplex PCR and randomly amplified polymorphic DNA (RAPD) were performed for the molecular determination. There was high molecular relatedness among 11 Korean isolates and 17 U.S. strains at 63% level. Additionally, distinct differentiation between pig and cattle isolates was determined. It implied that RAPD had a capacity to distinguish strains with different sources, however it could not discriminate among isolates according to their differences in the degree of virulence. In antimicrobial susceptibility tests, 45.5% of isolates showed antibiotic resistance to two or more antibiotics. Unlike the isolates from other countries, domestic isolates of E. coli O157:H7 was mainly resistant to ampicillin and tetracylines. In summary, the application of E. coli O157:H7 Rapid kit may be useful to detect E. coli O157:H7 due to its sensitivity and convenience. Moreover, combinational analysis of multiplex PCR together with RAPD can aid to survey the characteristics of isolates.

Keyword

Escherichia coli O157:H7; multiplex PCR; RAPD

MeSH Terms

Abattoirs
Adhesins, Bacterial/genetics
Animals
Cattle
Cercopithecus aethiops
Chickens
Escherichia coli O157/genetics/*isolation&purification
Escherichia coli Proteins/genetics
Feces/microbiology
Food Microbiology
Hemolysin Proteins/genetics
Korea
Meat/*microbiology
Phylogeny
Polymerase Chain Reaction/*methods
Random Amplified Polymorphic DNA Technique/*methods
*Reagent Kits, Diagnostic
Shiga-Like Toxin I/genetics
Shiga-Like Toxin II/genetics
Swine
United States
Vero Cells