Comparison of traditional and molecular typing methods of Escherichia coli O157.
An account is given using typing methods and detection of virulence genes of different serotypes of Escherichia coli isolated in Hungary. By hybridization using SLT-I and SLT-II probes and PCR method using stx1-2, eae and ehx primers we could differentiate O157 strains of different serotypes into eight (stx, eae, ehxA positive; stx, eae positive; stx, ehxA positive; stx positive; eae, ehxA positive; eae positive; ehxA positive; stx, eae, ehxA negative) types. The discriminatory power of phage typing proves to be much higher than that of the plasmid profile. RAPD typing with different primers could confirm or exclude the subtypes identity of the isolated E. coli O157 serotypes. Escherichia coli O157:HNM isolates could be sorted in six different phage types and six different RAPD types with ERIC-1, in five RAPD types with ERIC-2 and in seven types with M13 primers. Escherichia coli O157:H7 showed six different phage types and three RAPD types with ERIC-1 and ERIC-2 and five types with M13 primers. According to our results the standard PFGE protocol [32] gives the opportunity to differentiate epidemiologically independent but evolutionary related or unrelated isolates, but the practical value of PFGE method for epidemiological purposes must be confirmed by other or more restriction enzymes or using an other protocol. Summarizing our results we suggest the use of phage and RAPD typing and in doubtful cases the PFGE method.
Milch
Hedda
H
B. Johan National Center for Epidemiology, Gyáli út 2-6, H-1097 Budapest, Hungary.
Pászti
Judit
J
Gadó
I
I
Glatz
Katalin
K
Jakab
Melinda
M
D015373:Bacterial Typing Techniques / Q000379:methods*; D001434:Bacteriophage Typing / Q000379:methods; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D016521:Electrophoresis, Gel, Pulsed-Field; D019453:Escherichia coli O157 / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification / Q000472:pathogenicity; D005798:Genes, Bacterial; D006801:Humans; D006814:Hungary; D019105:Random Amplified Polymorphic DNA Technique / Q000379:methods; D012703:Serotyping; D014774:Virulence / Q000235:genetics
D017931:DNA Primers; D004269:DNA, Bacterial