Strategies for PCR based detection of Burkholderia pseudomallei DNA in paraffin wax embedded tissues.

ABSTRACT: Recently, several cases of melioidosis imported to Europe have been reported. The diagnosis of the acute or chronic infection remains challenging. This report describes an optimised protocol for fast and reliable DNA preparation for use in two different polymerase chain reaction (PCR) assays, namely: (1) a seminested PCR assay targeting a genus specific sequence of the ribosomal protein subunit 21 (rpsU) gene and (2) a nested PCR assay targeting the gene encoding the filament forming flagellin (fliC). Various strains of Burkholderia spp, strains of closely related genera, and spleen tissue samples of experimentally infected mice were investigated. The combination of PCR and sequencing of the amplicons resulted in high sensitivity and specificity. These procedures may allow rapid, sensitive

SUBMITTER: Hagen RM 

PROVIDER: S-EPMC1187279 | biostudies-literature | 2002 Dec

REPOSITORIES: biostudies-literature