title: Molecular and in vitro analysis of eight dengue type 2 viruses isolated from patients exhibiting different disease severities.
abstract: Potential genetic determinants of dengue virulence were studied by sequencing the entire genomes of eight dengue 2 virus strains isolated from patients exhibiting different disease severities during an epidemic season in northeastern Thailand in 1993. The isolates came from one dengue shock syndrome (ThNH-7/93), three dengue hemorrhagic fever, and four dengue fever patients. Phylogenetic analysis showed that the isolates belonged to the Southeast Asian genotype. The 3' noncoding regions showed distinctive secondary structures, with one specific structure for the isolate ThNH-7/93. Analysis of the predicted polyprotein showed several amino acid (aa) changes scattered mostly in the nonstructural region. Of 30 positions with aa changes, 7 were unique to the isolate ThNH-7/93 and 3 of those led to radical alterations in aa character. Several aa changes coincided with previous studies relating genome sequence and virulence. Minimal changes in computer-predicted protein secondary structures were observed. Infective particles in the inoculum for all isolates were approximately equal as measured by focus formation on BHK-21 cells, but this did not correlate with the number of plaques formed on LLC-MK2 cells. Isolates from patients that experienced secondary infection were shown to have significantly larger plaques than the isolates from primary infection patients.
authors.affiliation: Department of Virology, Nagasaki University, Japan. maloy@ep.tm.nagasaki-u.ac.jp
authors.forename: M N
authors.initials: MN
authors.lastname: Mangada
authors.forename: A
authors.initials: A
authors.lastname: Igarashi
mesh_terms: D000293:Adolescent; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D002648:Child; D006224:Cricetinae; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics; D003715:Dengue / Q000821:virology*; D003716:Dengue Virus / Q000145:classification / Q000235:genetics* / Q000472:pathogenicity; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D010641:Phenotype; D010802:Phylogeny; D010948:Viral Plaque Assay; D014774:Virulence
chemical_list: D004279:DNA, Viral