Comparison of real-time SYBR green dengue assay with real-time taqman RT-PCR dengue assay and the conventional nested PCR for diagnosis of primary and secondary dengue infection.

North American Journal of Medical Sciences
Damodar PaudelWatcharee Chokejindachai

Abstract

Dengue fever and dengue hemorrhagic fever are caused by dengue virus. Dengue infection remains a burning problem of many countries. To diagnose acute dengue in the early phase we improve the low cost, rapid SYBR green real time assay and compared the sensitivity and specificity with real time Taqman(®) assay and conventional nested PCR assay. To develop low cost, rapid and reliable real time SYBR green diagnostic dengue assay and compare with Taqman real-time assay and conventional nested PCR (modified Lanciotti). Eight cultured virus strains were diluted in tenth dilution down to undetectable level by the PCR to optimize the primer, temperature (annealing, and extension and to detect the limit of detection of the assay. Hundred and ninety three ELISA and PCR proved dengue clinical samples were tested with real time SYBR(®) Green assay, real time Taqman(®) assay to compare the sensitivity and specificity. Sensitivity and specificity of real time SYBR® green dengue assay (84% and 66%, respectively) was almost comparable to those (81% and 74%) of Taqman real time PCR dengue assay. Real time SYBR(®) green RT-PCR was equally sensitive in primary and secondary infection while real time Taqman was less sensitive in the secondary infe...Continue Reading

Citations

Feb 16, 2013·Food and Environmental Virology·Roberto A RodríguezCharles P Gerba
Apr 30, 2014·Sensors·Alvaro Díaz-BadilloFeliciano Sanchez-Sinencio
Oct 4, 2014·Tropical Animal Health and Production·Mohamed HarounNawal Mohamed Abdulla
Mar 15, 2015·Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology·Je-Hyoung KimHyun Park
Jan 21, 2015·Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology·Chonticha KlungthongStefan Fernandez
Dec 23, 2016·Expert Review of Molecular Diagnostics·Shamala Devi Sekaran, Hui Jen Soe
Dec 11, 2013·Arquivos De Gastroenterologia·Carlos Eduardo BeckerAna Beatriz Gorini da Veiga
Sep 30, 2015·The American Journal of Tropical Medicine and Hygiene·Yu YangRichard G Jarman
Nov 19, 2019·PLoS Neglected Tropical Diseases·Hideaki ShimizuTatsuo Shioda
Aug 9, 2020·Brazilian Journal of Microbiology : [publication of the Brazilian Society for Microbiology]·Erick Gustavo DorlassDanielle Bruna Leal Oliveira

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Methods Mentioned

BETA
enzyme-linked immunosorbent assay
PCR
ELISA
electrophoresis
Assay

Software Mentioned

SDS
Clustal X

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