A novel DANP-coupled hairpin RT-PCR for rapid detection of chikungunya virus
Autor(es): Chen Huixin, Takei Fumie, Koay Evelyn Siew-Chuan, Nakatani Kazuhiko, Chu Justin Jang Hann
Resumo: Chikungunya has re-emerged as an important arboviral infection of global health significance. Because of lack of a vaccine - 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 - cost-effective one-step RT-PCR assay, with high sensitivity - 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, - the specificity - sensitivity were 100% (95% CI, 80.0% to 100%) - 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.
Imprenta: The Journal of Molecular Diagnostics, v. 15, n. 2, p. 227-233, 2013
Identificador do objeto digital: 10.1016/j.jmoldx.2012.10.004
Descritores: Chikungunya virus - DNA ; Chikungunya virus - Genome ; Chikungunya virus - Molecular structure ; Chikungunya virus - Pathogenesis ; Chikungunya virus - RNA ; Chikungunya virus - Viral infections ; Chikungunya virus - Molecular methods ; Chikungunya virus - PCR detection ; Chikungunya virus - RT-PCR ; Chikungunya Virus - Virus ; Chikungunya virus - Molecular screening ; Chikungunya virus - Vaccine ; Chikungunya virus - Public health
Data de publicação: 2013
