Cloning and expression of the exposed portion of the envelop protein of the four dengue virus serotypes

Abstract

[Extract] Dengue fever is a growing public health concern affecting millions of people during annual epidemics in endemic areas. The symptoms of the disease include acute biphasic fever, rash and general malaise. However, sequential infection by different serotypes of the virus can sometimes result in dengue haemorrhagic fever or dengue shock syndrome, which can be fatal.\ud \ud In the absence of a cure for the disease, the control of dengue virus infection can only be achieved by effective public health measures. There is an urgent need for rapid, specific and easy-to-use dengue viral diagnosis tests in endemic areas. The diagnosis of dengue virus infection relies upon either the detection of viraemia during the first 3-5 days of the illness or by the detection of antibody thereafter. The detection of dengue infection by ELISA is preferred in most clinical laboratories because they are easy to do and large numbers of specimens can be processed. Until recently there has been no commercial ELISA. Most laboratories use in-house tests based on published protocols (Kuno, Gomez and Gubler 1991, Bundo and Igarashi 1985). These tests have used viral antigen derived from cell culture. With a view towards development of serotype specific ELISA tests, we wish to report the cloning of E genes encoding the estramembraneous portions of D1 to D4 and their high level expression using the T7 expression system (Studier et al. 1990). We also report the production of soluble domain of E protein from Dengue 2 in insect cells using recombinant Baculovirus

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