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PMID: 8290354 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

RAPD (arbitrary primer) PCR is more sensitive than multilocus enzyme electrophoresis for distinguishing related bacterial strains.

Nucleic acids research ·Vol. 21 ·No. 25 ·1993-12-25 ·Pages 5930-3

Wang G, Whittam TS, Berg CM, Berg DE

Abstract

The RAPD (random amplified polymorphic DNA) fingerprinting method, which utilizes low stringency PCR amplification with single primers of arbitrary sequence to generate strain-specific arrays of anonymous DNA fragments, was calibrated relative to the widely used, protein-based multilocus enzyme electrophoretic (MLEE) typing method. RAPD fingerprinting was carried out on five isolates from each of 15 major groups of Escherichia coli strains that cause diarrheal disease worldwide (75 isolates in all). Each group consisted of isolates that were not distinguishable from one another by MLEE typing using 20 diagnostic enzyme markers. In our RAPD tests, three or more distinct subgroups in each MLEE group were distinguished with each of five primers, and 74 of the 75 isolates were distinguished when data obtained with five primers were combined. Thus, RAPD typing is far more sensitive than MLEE typing for discriminating among related strains of a species. Despite their different sensitivities, the same general relationships among strains were inferred from MLEE and RAPD data. Thus, our results recommend use of the RAPD method for studies of bacterial population genetic structure and evolution, as well as for epidemiology.

MeSH Terms
Bacteria/classification,enzymology,genetics Bacterial Typing Techniques DNA Fingerprinting DNA Primers Electrophoresis Enzymes/chemistry Escherichia coli/classification,enzymology,genetics Phylogeny Polymerase Chain Reaction/methods Sensitivity and Specificity
Chemicals
DNA Primers Enzymes
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Wang G
Department of Molecular Microbiology, Washington University Medical School, St Louis, MO 63110.
Whittam T S
Berg C M
Berg D E
References (11)
11 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
Published
1993-12-25
Pages
5930-3
Language
English
Region
England
NLM ID
0411011
PMCID
PMC310476
Subset
IM
Grants
NIAID NIH HHS · AI00964 · United States
NIAID NIH HHS · AI24566 · United States
NHGRI NIH HHS · HG00563 · United States
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