The dengue virus subgroup of the flavivirus family is comprised of 4 distinct serotypes (type 1 to type 4). Intratypic variations have been detected within several serotypes by oligonucleotide fingerprint analysis and by virus neutralization. Dengue type 4 variants from the Caribbean recovered during 1981-1982 appear to be different from the dengue 4 prototype (H241) isolated in the Phillipines in 1956 as well as recent isolates from Southeast Asia. During the past two years, dengue type 4 was the most frequent cause of dengue hemorrhagic shock in Bangkok, in contrast to the earlier epidemic pattern in which dengue type 2 viruses were most often implicated. For these reasons it is important to define in molecular terms: (a) genetic stability and diversity of dengue 4 viruses isolated over a 30 year interval and (b) the possible involvement of specific virus strains, i.e., """"""""virulent viruses"""""""", in severe dengue disease. Two dengue type 4 variants, the prototype strain H241 and strain 2123 isolated from a hemorrhagic shock patient, were chosen for cloning and DNA sequence analysis. Several specific oligonucleotide primers corresponding to the established dengue 4 sequence (Dominica strain) were tested for their ability to prime reverse-transcription. At least one oligonucleotide effectively primed each dengue genomic RNA and yielded cDNA 5,000 nucleotides in length. These cDNA products will then be cloned by the procedure established earlier. Cloned DNA will then be analyzed to determine the sequence coding for the two protective antigens, the envelope glycoprotein and the NS1 nonstructural protein. Finally, sequences will be compared in an attempt to identify hypervariable regions in the E and NS1 glycoproteins.
