Dengue typing assay based on real-time PCR using SYBR green I

Typing of dengue virus is crucial for the epidemiology and pathogenesis of dengue virus infection. Hence, highly sensitive and accurate diagnostic tools are essential. The purpose of this study was to identify all four types of dengue virus based on the 3'-untranslated region of the virus by melting curve analysis and real-time PCR using SYBR Green I. The types obtained by this method were compared with the results of direct sequencing of 39 serum or plasma samples of patients with clinical dengue infection that included a positive tourniquet test, thrombocytopenia and positive dengue IgM antibody. The accuracy of typing by melting curve analysis was 97.4%. In conclusion, real-time PCR and melting curve analysis using one single-primer pair were shown to be highly efficient for clear detection and typing of dengue virus in clinical specimens. This method there for represents a simple, sensitive, specific, rapid and economic method, which will be essential for epidemiological studies of dengue virus infection. (c) 2005 Elsevier B.V. All rights reserved.