Multiplex PCR assay simplifies serotyping and sequence typing of Listeria monocytogenes associated with human outbreaks.
Listeria monocytogenes serotypes 1/2a and 4b are responsible for the majority of cases of human listeriosis worldwide. In this study, a multiplex PCR assay was developed to allow rapid identification and easily interpretable differentiation of serotypes 1/2a and 4b from other serotypes of L. monocytogenes by simultaneously targeting two virulence genes (inlB and inlC) and two serotype-specific genes (ORF2372 and Imo0171). A subsequent gel extraction and sequence typing analysis of the highly polymorphic intragenic regions in inlB and inlC simplified a previously developed multi-virulence-locus sequence typing scheme and provided discriminatory power for subtyping L. monocytogenes similar to pulsed-field gel electrophoresis analysis.
Zhang
Wei
W
Department of Food Science, Pennsylvania State University, University Park, Pennsylvania 16802-2504, USA. wzhang@cdc.gov
Knabel
Stephen J
SJ
D015373:Bacterial Typing Techniques; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis*; D004196:Disease Outbreaks / Q000517:prevention & control; D005516:Food Microbiology; D006801:Humans; D008089:Listeria monocytogenes / Q000145:classification* / Q000302:isolation & purification*; D008088:Listeriosis / Q000453:epidemiology / Q000382:microbiology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D012703:Serotyping; D014774:Virulence / Q000235:genetics
D017931:DNA Primers; D004269:DNA, Bacterial