Abstract
BackgroundDengue is emerging as a major public health concern in many parts of the world. The development of a one-step, single tube, rapid, and multiplex reverse transcription polymerase chain reaction (M-RT-PCR) for simultaneous detection and typing of dengue virus using serotype specific primers during acute phase of illness is reported.ResultsAn optimal assay condition with zero background was established having no cross-reaction with closely related members of flavivirus (Japanese encephalitis, West Nile, Yellow fever) and alphavirus (Chikungunya). The feasibility of M-RT-PCR assay for clinical diagnosis was validated with 620 acute phase dengue patient sera samples of recent epidemics in India. The comparative evaluation vis a vis conventional virus isolation revealed higher sensitivity. None of the forty healthy serum samples screened in the present study revealed any amplification, thereby establishing specificity of the reported assay for dengue virus only.ConclusionThese findings clearly suggested that M-RT-PCR assay reported in the present study is the rapid and cost-effective method for simultaneous detection as well as typing of the dengue virus in acute phase patient serum samples. Thus, the M-RT-PCR assay developed in this study will serve as a very useful tool for rapid diagnosis and typing of dengue infections in endemic areas.
Highlights
IntroductionThe development of a one-step, single tube, rapid, and multiplex reverse transcription polymerase chain reaction (M-Reverse transcription polymerase chain reaction (RT-PCR)) for simultaneous detection and typing of dengue virus using serotype specific primers during acute phase of illness is reported
Dengue is emerging as a major public health concern in many parts of the world
In dengue infection, serotyping is very important because of the fact that secondary infection with a heterologous serotype often leads to life threatening dengue haemorrhagic fever (DHF) and dengue shock syndrome (DSS) [4]
Summary
The development of a one-step, single tube, rapid, and multiplex reverse transcription polymerase chain reaction (M-RT-PCR) for simultaneous detection and typing of dengue virus using serotype specific primers during acute phase of illness is reported. Dengue is the most important mosquito borne viral infection and is prevalent in most parts of the tropics. One hundred million cases of dengue fever (DF) and 450,000 cases of dengue hemorrhagic fever/dengue shock syndrome (DHF/DSS) are reported annually [1,2,3]. The most challenging problem associated with dengue is patient management, which is possible through rapid diagnosis of early infection. In dengue infection, serotyping is very important because of the fact that secondary infection with a heterologous serotype often leads to life threatening dengue haemorrhagic fever (DHF) and dengue shock syndrome (DSS) [4]
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