Establishment of one-step SYBR green-based real time-PCR assay for rapid detection and quantification of chikungunya virus infection | Ho et al. Virology Journal 2010 7 13 http content 7 1 13 VIROLOGY JOURNAL SHORT REPORT Open Access Establishment of one-step SYBR green-based real time-PCR assay for rapid detection and quantification of chikungunya virus infection Phui San Ho1 Mary Mah Lee Ng2 Justin Jang Hann Chu2 Abstract Chikungunya virus CHIKV is a mosquito-borne alphavirus and one of the prevalent re-emerging arbovirus in tropical and subtropical regions of Asia Africa and Central and South America. It produces a spectrum of illness ranging from inapparent infection to moderate febrile illness as well as severe arthralgia or arthritis affecting multiple joints. In this study a quantitative one-step real-time SYBR Green-based RT-PCR system for the non-structural protein 2 nsP2 of CHIKV that can quantify a wide range of viral RNA concentrations was developed. Comparisons between the conventional semi-quantitative RT-PCR assay immunofluorescence detection method and the one-step SYBR Green-based RT-PCR assay in the detection of CHIKV infection revealed much rapid and increase sensitivity of the latter method. Furthermore this newly developed assay was validated by in vitro experiments in which ribavirin a well-known RNA virus inhibitor showed a dose-dependent inhibition of virus replication on cells that was assessed by viral infectivity and viral RNA production. Our results demonstrate the potential of this newly developed one-step SYBR Green I-based RT-PCR assay may be a useful tool in rapid detection of CHIKV and monitoring the extent of viral replication possibly in patients samples. Findings Chikungunya virus disease or chikungunya fever is a viral disease transmitted to humans by the bite of infected Aedes_aegypti yellow fever mosquito and Aedes albopictus tiger mosquito 1 . Chikungunya virus CHIKV is a member of the genus alphavirus in the family Togaviridae 2 . CHIKV contains a non-seg-mented monomeric positive-sense RNA genome that is approximately 12 Kb. CHIKV genome
