Rapid and Real-time Assays for Detection and Quantification of Chikungunya Virus

MM Parida; SR Santhosh; PK Dash; PV Lakshmana Rao

Disclosures

Future Virology. 2008;3(2):179-192. 

In This Article

Real-time Assays

Recent advances in PCR amplification methods, which hold a great deal of promise for highly specific and sensitive virus detection assays, combine PCR amplification with fluorescently-labeled virus-specific probes able to detect amplified DNA during the amplification reaction. The fluorescent chemistry coupled with advanced optical detectors renders such techniques more sensitive than conventional gel-based PCR. Over the past 2 years, several real-time PCR assays have been developed to address the requirement of reliable detection systems for early detection as well as quantification of viral load in the acute phase of illness. The scope of the present review is to appraise the recent advances in the field of molecular diagnosis of CHIKV with reference to real-time assay systems for rapid and reliable detection in addition to quantification of viral load in the early stage of infection.

Real-time assays have many advantages over conventional PCR methods, including rapidity, quantitative measurement, lower contamination rate, higher sensitivity, higher specificity and ease of standardization.[23] The development of real-time PCR has transferred quantitation of target nucleic acids from the pure research laboratory to the clinical diagnostic laboratory. In addition, real-time PCR automates the laborious process of amplification by quantifying reaction products for each sample in every cycle. Data analysis, including standard curve generation and copy number calculation, is performed automatically. Real-time RT-PCR assays targeting the E1 immunodominant gene of the CHIKV employing both TaqMan® and SYBR® Green I chemistry have been reported.

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