Open AccessEvaluation of a PCR Melting Profile Technique for Bacterial Strain Differentiation
Author(s) -
Beata Krawczyk,
Alfred Samet,
Justyna Leibner,
Anna Śledzińska,
Józef Kur
Publication year - 2006
Publication title -
journal of clinical microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.349
H-Index - 255
eISSN - 1070-633X
pISSN - 0095-1137
DOI - 10.1128/jcm.00052-06
Subject(s) - typing , pulsed field gel electrophoresis , gel electrophoresis , polymerase chain reaction , biology , molecular epidemiology , microbiology and biotechnology , genotype , genetics , gene
In search of an effective DNA typing technique for hospital epidemiology use, the performance and convenience of a PCR melting profile (PCR MP) technique based on using low denaturation temperatures during ligation-mediated PCR (LM PCR) of bacterial DNA was tested. A number ofEscherichia coli isolates from patients of the Clinical Hospital in Gdańsk, Poland, were examined. We found that the PCR MP technique is a rapid method that offers good discriminatory power and excellent reproducibility and may be applied for epidemiological studies. The usefulness of the PCR MP for molecular typing was compared with the pulsed-field gel electrophoresis method, which is currently considered the gold standard for epidemiological studies of isolates recovered from patients and the environment. Clustering of PCR MP fingerprinting data matched pulsed-field gel electrophoresis data. The features of the PCR MP technique are discussed in comparison with conventional methods. Data presented here demonstrate the complexity of the epidemiological situation concerningE. coli that may occur in a hospital.