Genetic and biological differentiation of dengue 3 isolates obtained from clinical cases in Java, Indonesia, 1976?1978

Abstract

Previous epidemiological, virological and clinical studies have documented a series of outbreaks of dengue fever and dengue haemorrhagic fever/dengue shock syndrome which occurred in Java, Indonesia in 1976-1978. In the current study we compare growth characteristics in cell culture, and nucleotide sequence data for the viral prM and E genes, of five low passage DEN-3 isolates obtained during these epidemics from clinically defined cases. All isolates had the same passage history: human sera were passed twice in mosquitoes and three times in a mosquito cell line (Aedes albopictus, C 6/36 cells). Growth differences were observed between individual isolates in Vero cells; growth differences were not observed in C 6/36 cells. Nucleotide sequencing of the prM and E gene region indicated that no two isolates were identical (sequence divergence ranged from 0.4 to 1.6% in pairwise comparisons) but that they were closely enough related to present a single genetic type. There were one or two differences in deduced amino acid sequence in E between isolates. Differences were at residues 65, 187, 298 or 443. One isolate differed from all others at residue 16 in the M protein. No relationship was apparent between the amino acid sequence of M or E and the nature of the disease profile, the year of isolation or the geographic region of isolation. The isolates showed 3.5 to 4.4% nucleotide sequence divergence from the highly-adapted H 87 prototype, isolated in the Philippines in 1956. The isolates showed a total of twelve common amino acid differences in prM and E proteins from H 87. Ten of these twelve residues were at positions which differed between the four dengue serotypes. Two differences (at residues 37 in M and 293 in E) were at positions which are conserved in sequence between the four dengue serotypes. The data are discussed in relation to the dengue outbreaks in Java in the period 1976-1978.

Clinical and virological characteristics of dengue in Surabaya, Indonesia

“…The AA substitutions were shared by isolates causing both DF and DHF (Fig 6). Previous studies have also observed similar findings and have found no reproducible genetic differences related to disease severity [69–72]. However, we are aware that only E protein AA sequences were compared in this study.…”

Comparative Analysis of NS3 Sequences of Temporally Separated Dengue 3 Virus Strains Isolated from Southeast Asia

“…This pic ture concurs with the data of other workers who studied other dengue 3 viral genes [16][17][18]. 256 Chow/Seah/Chan NS3 Sequences of Dengue 3 Virus Strains Interestingly, the strains from Malaysia and Singapore isolated in the more recent outbreaks of the late 1980s and early 1990s were more closely related to the Thai cluster, implying their evolution from the latter.…”

“…The correlation of some of our findings derived from the more 'stable' NS3 gene compared with those of work ers elsewhere who studied other relatively more 'variable' dengue 3 virus genes such as the envelope, premembrane and membrane sequences [16][17][18] verify the validity of our data. In addition, the useful interpretation of new evo lutionary and/or epidemiologic links exhibits the feasibili ty of exploiting NS3 base differences between strains for constructing informative phylogenetic trees.…”

Dengue virus infection rate in field populations of female Aedes aegypti and Aedes albopictus in Singapore

“…1999; Holmes & Burch 2000). Differences among dengue of the same type within the same geographical region was previously reported by Lee et al . (1993), Attatippaholkun et al .…”