Skip to main content

Molecular Strategies for the Detection, Identification, and Differentiation between Enteroinvasive Escherichia coli and Shigella spp.

Buy Article:

$37.00 plus tax (Refund Policy)

Abstract:

A strategy for the detection, identification, and differentiation of enteroinvasive Escherichia coli (EIEC) and Shigella spp. has been developed. The strategy includes (i) a multiplex PCR for the amplification of two virulence genes, i.e., iuc (222 bp) and ipaH (629 bp); (ii) amplification of the ial gene (a 1,038-bp amplicon) located within a large plasmid; and (iii) restriction fragment length polymorphism (RFLP) of the ial gene amplicon. The multiplex PCR provided three patterns. Pattern 1 (iuc /ipaH +) was found in 10 (67%) of 15 EIEC strains tested, pattern 2 (iuc +/ipaH ) in only 2 (4.4%) of 46 non-EIEC isolates, whereas pattern 3 (iuc +/ipaH +) was observed in all Shigella spp. and also in 5 (33%) of 15 EIEC strains tested. The pattern 3 EIEC strains were all positive for the ial gene. The PCR-RFLP of the ial gene amplicon using the endonuclease Ac/I was used to differentiate Shigella spp. from the EIEC strains that belonged to pattern 3. The ial gene was present in 21 (38%) of 56 and 6 (40%) of 15 Shigella spp. and EIEC strains tested, respectively. The PCR-RFLP of the ial gene amplicon divided the strains in two types. Type 1 did not contain the restriction enzyme site and was found in 6 (100%) of 6 EIEC strains, 4 (80%) of 5 Shigella boydii, and 4 (100%) of 4 Shigella dysenteriae strains tested. Type 2, which gave two fragments of 286 and 752 bp, was observed in 5 (83%) of 6 Shigella flexneri strains and 6 (100%) of 6 Shigella sonnei strains. Detection, identification, and differentiation of Shigella spp. and EIEC were achieved by analyses of the PCR patterns and RFLP types. To our knowledge, this is the first study to demonstrate a simple and rapid method for detecting, identifying, and differentiating, at the molecular level, Shigella spp. and EIEC strains. This method will have tremendous utility as an epidemiological tool and in helping to develop policies, risk assessments, and national and international methods for Shigella spp.

Document Type: Research Article

Affiliations: Health Products and Food Branch, Bureau of Microbial Hazards, Health Canada, Sir Frederick G. Banting Research Centre, Tunney's Pasture, P. L. 2204A2, Ottawa, Ontario, Canada K1A 0L2

Publication date: February 1, 2005

More about this publication?
  • IAFP Members with personal subscriptions to JFP Online: To access full-text JFP or JMFT articles, you must sign-in in the upper-right corner using your Ingenta sign-in details (your IAFP Member Login does not apply to this website).

    The Journal of Food Protection (JFP) is a refereed monthly publication. Each issue contains scientific research and authoritative review articles reporting on a variety of topics in food science pertaining to food safety and quality. The Journal is internationally recognized as the leading publication in the field of food microbiology with a readership exceeding 11,000 scientists from 70 countries. The Journal of Food Protection is indexed in Index Medicus, Current Contents, BIOSIS, PubMed, Medline, and many others.

    Print and online subscriptions are available to IAFP Members and institutional subscribers. IAFP Members with a subscription to JFP Online will have access to all available JFP and JMFT content. Online visitors who are not IAFP Members or journal subscribers will be charged on a pay-per-view basis. Membership and subscription information is available at www.foodprotection.org.
  • Information for Authors
  • Submit a Paper
  • Subscribe to this Title
  • Membership Information
  • Information for Advertisers
  • Ingenta Connect is not responsible for the content or availability of external websites

Access Key

Free Content
Free content
New Content
New content
Open Access Content
Open access content
Partial Open Access Content
Partial Open access content
Subscribed Content
Subscribed content
Free Trial Content
Free trial content
Cookie Policy
X
Cookie Policy
Ingenta Connect website makes use of cookies so as to keep track of data that you have filled in. I am Happy with this Find out more