Evaluation of pre-analytical variables in the quantification of dengue virus by real-time polymerase chain reaction.

ABSTRACT: An accurate molecular diagnosis for viral pathogens is highly dependent on pre-analytical procedures. The efficiencies of two viral RNA extraction methods (liquid phase partition and silica-based adsorption chromatography) and the effects of handling and storage on the stability of RNA isolated from dengue virus (DENV) were studied. Viral RNA extracted from spiked sera or clinical samples characterized with DENV infection were quantified by TaqMan real-time PCR. The presence of high serum proteins severely affected the recovery of DENV RNA by the liquid phase partition, but not the silica-based method. The recovery with Trizol liquid phase partition method was significantly improved by a concomitant addition of a co-precipitant and the reduction of sera proteins, resulting in recoveries si

SUBMITTER: Anwar A 

PROVIDER: S-EPMC2765752 | biostudies-literature | 2009 Nov

REPOSITORIES: biostudies-literature