Geographical clusters of dengue virus type 2 isolates based on analysis of infected cell RNA by the chemical cleavage at mismatch method

B. Lin, R. G.H. Cotton, D. W. Trent, P. J. Wright

Research output: Contribution to journalArticle

5 Scopus citations

Abstract

Genetic variation in 12 strains of dengue virus type 2, isolated from several epidemic areas in different years, was studied by chemical cleavage at mismatched cytosine in DNA:RNA heteroduplexes. End-labelled cDNA probes derived from the E and NS2A genes of the New Guinea C strain were hybridized to total RNA extracted from cells infected by individual isolates. Following modification of mismatched cytosine by hydroxylamine and nucleic acid strand cleavage by piperidine, the resulting fragments of radiolabelled probe were analysed by electrophoresis and autoradiography. The patterns of bands generated corresponded to the geographical groupings of the isolates. Thus this method is suitable in epidemiological studies for rapidly surveying a large number of isolates for genetic variation in a particular gene of interest.

Original languageEnglish (US)
Pages (from-to)205-218
Number of pages14
JournalJournal of Virological Methods
Volume40
Issue number2
DOIs
StatePublished - Nov 1992
Externally publishedYes

Keywords

  • Dengue virus type 2
  • Hydroxylamine
  • Topotype

ASJC Scopus subject areas

  • Virology

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