RAPD (arbitrary primer) PCR is more sensitive than multilocus enzyme electrophoresis for distinguishing related bacterial strains.
AUTOR(ES)
Wang, G
RESUMO
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.
ACESSO AO ARTIGO
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=310476Documentos Relacionados
- CODEHOP (COnsensus-DEgenerate Hybrid Oligonucleotide Primer) PCR primer design
- Methods of multilocus enzyme electrophoresis for bacterial population genetics and systematics.
- Screening for overlapping bacterial artificial chromosome clones by PCR analysis with an arbitrary primer
- Piliated Bacteroides fragilis strains adhere to epithelial cells and are more sensitive to phagocytosis by human neutrophils than nonpiliated strains.
- Arbitrary primer polymerase chain reaction, a powerful method to identify Bacillus thuringiensis serovars and strains.