Rapid and real-time assays for detection and quantification of Chikungunya virus

Abstract

Chikungunya virus (CHIKV), a member of the Alphavirus genus, is a considerable public health concern in Southeast Asian and African countries. Despite the fact that CHIKV resurgence is associated with epidemics of unprecedented magnitude, only a few specific serological and molecular diagnostic tools are available. CHIKV diagnosis is essentially based on virus isolation, ELISA and reverse transcription (RT)-PCR assays. RT-PCR is the method of choice for the early detection and confirmation of virus in clinical samples. Further advancement in terms of rapid, reliable detection and quantification with improved sensitivity has been accomplished through development of both fully automated TaqMan® and SYBR® Green I-based real-time RT-PCR assays. In addition, another simple, rapid, novel and cost-effective isothermal gene amplification method known as RT loop-mediated isothermal amplification (RT-LAMP) has also been reported for the early detection and quantification of viral genomes in acute-phase patient serum samples. Of notable importance is the substantial reduction in time required for the confirmation of results by RT-LAMP assay (30 min) and monitoring of amplification by SYBR Green I dye-mediated naked-eye visualization. These findings demonstrate that the real-time RT-PCR and RT-LAMP assays have potential applications in clinical diagnostics owing to simultaneous detection and quantification of CHIKV in acute phase patient serum samples.