Development and Clinical Evaluation of A PCR Assay Targeting the Metalloprotease Gene (mprA) of B. pseudomallei

Abstract

A PCR assay targeting the metalloprotease gene (mprA) of Burkholderia pseudomallei was developed for the specific detection of this organism in pure cultures and clinical samples. All other closely related organisms including B. mallei the causative agent of glanders, and B. thailandensis tested negative. Burkholderia pseudomallei DNA was successfully amplified from paraffin-embedded lung tissue of a camel with a generalized B. pseudomallei infection. The developed PCR assay can be used as a simple tool for the specific and sensitive detection of B. pseudomallei.

Discrimination ofBurkholderia mallei/pseudomalleifromBurkholderia thailandensisby sequence comparison of a fragment of the ribosomal protein S21 (rpsU) gene

“…Due to the lacking reliability of phenotypic methods for the discrimination within the Burkholderia genus, various molecular methods have been developed, including conventional, real-time, and multiplex polymerase chain reactions (PCRs), loop-mediated isothermal amplification (LAMP) procedures, sequence-based approaches, DNA micro-arrays, and fluorescence in situ hybridization [28][29][30][31][32][33][34][35].…”

Comparison of TaqMan PCR Assays for Detection of the Melioidosis Agent Burkholderia pseudomallei in Clinical Specimens

“…These include DNA microarrays (25), gene sequencing (15,32), isothermal DNA amplification (7), and real-time PCR assays targeting specific regions of the B. pseudomallei genome (2,5,(19)(20)(21)(26)(27)(28)(29)(30)) (see Table S1 in the supplemental material).…”

Burkholderia mallei : Glanders

“…pseudomallei complex based on the 16S rRNA and the fliC gene [13][14][15][16][17][18][19]. Other methods targeting the flip gene [20,21], the metalloprotease gene [22], the type three secretion system [23], and the bimA ma gene [24] can be used to confirm the isolate to be B. mallei. A newly established microarray-based method allows the fast and accurate identification and differentiation of B. mallei and B. pseudomallei using at least four independent genetic markers including the16S rRNA gene, fliC -and motB gene [25].…”