Genotyping of clinical Serratia marcescens isolates: a comparison of PCR-based methods.
The polymerase chain reaction (PCR)-based procedures of randomly amplified polymorphic DNA (RAPD) and repetitive element (RE)-based PCR were used to amplify total DNA prepared from each of 62 clinical Serratia marcescens isolates. Three different random primers, designated 1060, 1254 and 1283, were used individually in RAPD-PCR. Primers representing enterobacterial repetitive intergenic consensus (ERIC) sequences, extragenic palindromic (REP) elements, and polymorphic GC-rich repetitive sequences (PGRS) constituted the repetitive element-PCR. We were able to generate 40, 40 and 58 genotypic groupings using the 1060, 1254 and 1283 RAPD primers, respectively. Using the ERIC, REP and PGRS primers, 19, 54 and 60 unique genotypic profiles were yielded, respectively. The PGRS primers, which were developed to amplify GC-rich repetitive sequences in the genome of Mycobacteria, were the most discriminatory. These data indicate that both of these PCR-based approaches are a valid means of discriminating strain differences among isolates of S. marcescens and the amount of differentiation depends on the primer used. These techniques should prove useful for routine surveillance or in examining outbreaks of S. marcescens in clinical settings.
Patton
T G
TG
Department of Biological Sciences, 1200 Commercial, Emporia State University, Emporia, KS 66801, USA.
Katz
S
S
Sobieski
R J
RJ
Crupper
S S
SS
D015373:Bacterial Typing Techniques; D004269:DNA, Bacterial / Q000032:analysis; D005838:Genotype; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D019105:Random Amplified Polymorphic DNA Technique; D016868:Serratia Infections / Q000382:microbiology*; D012706:Serratia marcescens / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification
D004269:DNA, Bacterial