ArticleThe Journal of molecular diagnostics : JMD2009
Evaluation of pre-analytical variables in the quantification of dengue virus by real-time polymerase chain reaction.
Article in The Journal of molecular diagnostics : JMD, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 9 papers.
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Who cites it
9 citing papers in PubMed.
- Assessment of Strategies for Preserving Swine Viral RNA Targets in Diagnostic Specimens.Microorganisms · 2024Review
- Effect of extrinsic factors on the detection of PRRSV and a porcine-specific internal sample control in serum, oral fluid, and fecal specimens tested by RT-rtPCR.Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc · 2023Article
- Urinary genome detection and tracking of Hantaan virus from hemorrhagic fever with renal syndrome patients using multiplex PCR-based next-generation sequencing.PLoS neglected tropical diseases · 2021Article
- Dengue diagnostics: serious inaccuracies are likely to occur if pre-analytical conditions are not strictly followed.Memorias do Instituto Oswaldo Cruz · 2021Article
- SARS-CoV-2 RNA detected in blood products from patients with COVID-19 is not associated with infectious virus.Wellcome open research · 2020Article
- Isolation and molecular characterization of dengue virus clinical isolates from pediatric patients in New Delhi.International journal of infectious diseases : IJID : official publication of the International Society for Infectious Diseases · 2019Article
- An RT-PCR panel for rapid serotyping of dengue virus serotypes 1 to 4 in human serum and mosquito on a field-deployable PCR system.PloS one · 2019Article
- Quality assurance of RNA expression profiling in clinical laboratories.The Journal of molecular diagnostics : JMD · 2012Review
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Authors and funding
5 authors.
Funding
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 similar to that of the silica-based methods. Repeated freeze-thaw cycles did not affect the recovery of viral RNA. While intact DENV was found to be stable in serum for up to 2 hour at 25 degrees C, recovery of viral RNA from sera stored in the lysis/binding buffer was stable for up to 5 days. These data indicate that the choice of viral RNA extraction methods, the conditions for handling, and storing of clinical sera critically affect the quantification of viral nucleic acid from clinical samples. This will impact the accuracy and reproducibility of DENV diagnosis by PCR-based assays.
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