Evaluation of a PCR melting profile technique for bacterial strain differentiation.
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 of Escherichia 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 concerning E. coli that may occur in a hospital.
Krawczyk
Beata
B
Gdańsk University of Technology, Department of Microbiology, and Department of Clinical Microbiology, Clinical Hospital No. 1, Gdańsk, Poland.
Samet
Alfred
A
Leibner
Justyna
J
Sledzińska
Anna
A
Kur
Józef
J
D000328:Adult; D015373:Bacterial Typing Techniques; D003428:Cross Infection / Q000382:microbiology; D016172:DNA Fingerprinting / Q000379:methods*; D004269:DNA, Bacterial / Q000032:analysis* / Q000235:genetics; D016521:Electrophoresis, Gel, Pulsed-Field; D016021:Epidemiologic Studies; D004926:Escherichia coli / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D004927:Escherichia coli Infections / Q000382:microbiology; D006761:Hospitals; D006801:Humans; D007297:Inpatients; D017720:Molecular Epidemiology / Q000379:methods; D011044:Poland; D016133:Polymerase Chain Reaction / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D044366:Transition Temperature
D004269:DNA, Bacterial