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Title: A novel DANP-coupled hairpin RT-PCR for rapid detection of Chikungunya virus
Authors: Chen, H 
Takei, F
Koay, ESC 
Nakatani, K
Chu, JJH 
Keywords: Alphavirus Infections
Chikungunya virus
DNA Primers
Inverted Repeat Sequences
Reproducibility of Results
Reverse Transcriptase Polymerase Chain Reaction
Sensitivity and Specificity
Issue Date: 1-Mar-2013
Publisher: Elsevier BV
Citation: Chen, H, Takei, F, Koay, ESC, Nakatani, K, Chu, JJH (2013-03-01). A novel DANP-coupled hairpin RT-PCR for rapid detection of Chikungunya virus. Journal of Molecular Diagnostics 15 (2) : 227-233. ScholarBank@NUS Repository.
Abstract: Chikungunya has re-emerged as an important arboviral infection of global health significance. Because of lack of a vaccine and effective treatment, rapid diagnosis plays an important role in early clinical management of patients. In this study, we have developed a novel molecular diagnostic platform that ensures a rapid and cost-effective one-step RT-PCR assay, with high sensitivity and specificity, for the early detection of the Chikungunya virus (CHIKV). It uses 2,7-diamino-1,8-naphthyridine derivative (DANP)-labeled cytosine-bulge hairpin primers to amplify the nsP2 region of the CHIKV genome, followed by measurement of the fluorescence emitted from DANP-primer complexes after PCRs. The detection limit of our assay was 0.01 plaque-forming units per reaction of CHIKV. Furthermore, the HP-nsP2 primers were highly specific in detecting CHIKV, without any cross-reactivity with the panel of RNA viruses validated in this study. The feasibility of the DANP-coupled hairpin RT-PCR for clinical diagnosis was evaluated using clinical serum samples from CHIKV-infected patients, and the specificity and sensitivity were 100% (95% CI, 80.0% to 100%) and 95.5% (95% CI, 75.1% to 99.8%), respectively. These findings confirmed its potential as a point-of-care clinical molecular diagnostic assay for CHIKV in acute-phase patient serum samples. Copyright © 2013 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved.
Source Title: Journal of Molecular Diagnostics
ISSN: 15251578
DOI: 10.1016/j.jmoldx.2012.10.004
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