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Comparison of Techniques for Extracting Viral RNA in Plasma for Dengue Diagnosis by Reverse Transcriptase-PCR

1, 3, 4

  1. 1Department of Medical Technology, Far Eastern University, Manila, PHILIPPINES.
  2. 2The Graduate School, University of Santo Tomas, Manila, Philippines 3Research & Biotechnology Division, St. Luke’s Medical Center, Quezon City, PHILIPPINES.
  3. 3The Graduate School, University of Santo Tomas, Manila, Philippines
  4. 4Research & Biotechnology Division, St. Luke’s Medical Center, Quezon City, PHILIPPINES.

CORRESPONDENCE

Aaron Jan S. Palmares

Department of Medical Technology, Far Eastern University, Manila, PHILIPPINES.

apalmares@feu.edu.ph

Received: 13-06-2017; Accepted: 29-10-2017.

Volume 6, Issue 3 · pp. 396-400 · 2017 · DOI:

ABSTRACT

This study aims to determine if the rate of RT-PCR detection of Dengue virus from clinical plasma would vary between the RNA extracts of QIAamp® Viral RNA and TRIzol® LS in the first week of Dengue infection. Plasma samples from 31 individuals clinically suspected of being infected with dengue virus in day 1 to 7 of fever onset were extracted for RNA using the protocol of QIAamp® Viral RNA and TRIzol® LS. The paired RNA extracts of plasma samples were analyzed for the presence of Dengue virus RNA using RT-PCR. Out of 31 samples, a significantly higher rate of RT-PCR detection was obtained with QIAamp® than TRIzol® (74% vs. 48%, p=0.039). In comparison to Dengue NS1 antigen positivity, a significantly lower rates of RT-PCR detection was obtained with TRIzol® (77% vs. 48%, p=0.035), while there was no significant difference with QIAamp® (77% vs. 74%, p=1.000). In comparison to Dengue IgM/IgG antibody positivity, the rates of RT-PCR detection with QIAamp® was significantly higher (42% vs. 74%, p=0.031) while there was no significant difference with TRIzol® (42% vs. 48%, p=0.832).These results suggest that RNA extraction using QIAamp® Viral RNA provides more sufficiently pure RNA template for a conventional RT-PCR than TRIzol® LS.

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    Palmares, A. J. S., Chan, V. F., & Baclig, M. O. (2017). Comparison of Techniques for Extracting Viral RNA in Plasma for Dengue Diagnosis by Reverse Transcriptase-PCR. Asian Journal of Biological and Life Sciences, 6(3), 396-400.

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    © 2017 Author(s)

    This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International License, which permits unrestricted use, distribution and reproduction in any medium, provided the original author and source are credited. CC BY 4.0