A simple one‐step real‐time RT‐PCR for diagnosis of dengue virus infection

Abstract

Dengue is the most important arbovirus disease in tropical and sub‐tropical countries, and can be caused by infection with any of the four‐dengue virus (DENV) serotypes. Infection with DENV can lead to a broad clinical spectrum, ranging from sub‐clinical infection or an influenza‐like disease known as dengue fever (DF) to a severe, sometimes fatal, disease characterized by hemorrhage and plasma leakage that can lead to shock, known as dengue hemorrhagic fever/dengue shock syndrome (DHF/DSS). The diagnosis of dengue is routinely accomplished by serologic assays, such as IgM and IgG ELISAs, as well as HI tests, analyzing serum samples obtained from patients with at least 7 days of symptoms onset. These tests cannot be used for diagnosis during the early symptomatic phase. In addition, antibodies against dengue are broad reactive with other flaviviruses. Therefore, a specific diagnostic method for acute DENV infection is of great interest. In that sense, the real‐time RT‐PCR has become an important tool that can be used for early and specific detection of dengue virus genome in human serum samples. This study describes a simple, specific, and sensitive real‐time RT‐PCR for early diagnosis of dengue virus infection. J. Med. Virol. 80:1426–1433, 2008.