Combined detection and genotyping of Chikungunya virus by a specific reverse transcription-polymerase chain reaction

J Med Virol. 2002 Jul;67(3):370-4. doi: 10.1002/jmv.10085.


A reverse transcription-polymerase chain reaction (RT-PCR) was developed for the detection of Chikungunya virus infection. Based on the nonstructural protein 1 (nsP1) and glycoprotein E1 (E1) genes of Chikungunya, two primer sets were designed. Total RNA were extracted from the cell culture fluid of Aedes albopictus C6/36 cells inoculated with the S27 prototype virus, isolated in Tanzania in 1953, and the Malaysian strains (MALh0198, MALh0298, and MALh0398), isolated in Malaysia in 1998. For both sets of RNA samples, the expected 354- and 294-base pair (bp) cDNA fragments were amplified effectively from the nsP1 and E1 genes, respectively. Phylogenetic analysis was conducted for the Malaysian strain and other virus strains isolated from different regions in the world endemic for Chikungunya, using partial E1 gene sequence data. The Malaysian strains isolated during the epidemics of 1998 fell into a cluster with other members of the Asian genotype.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aedes
  • Alphavirus Infections / diagnosis*
  • Alphavirus Infections / epidemiology*
  • Alphavirus Infections / virology
  • Animals
  • Cells, Cultured
  • Chikungunya virus / classification*
  • Chikungunya virus / genetics
  • Chikungunya virus / isolation & purification*
  • Disease Outbreaks*
  • Genotype
  • Humans
  • Malaysia / epidemiology
  • Molecular Sequence Data
  • Phylogeny
  • Reverse Transcriptase Polymerase Chain Reaction*
  • Sensitivity and Specificity
  • Sequence Analysis, DNA
  • Viral Envelope Proteins / chemistry
  • Viral Envelope Proteins / genetics
  • Viral Envelope Proteins / metabolism
  • Viral Nonstructural Proteins / chemistry
  • Viral Nonstructural Proteins / genetics
  • Viral Nonstructural Proteins / metabolism


  • Viral Envelope Proteins
  • Viral Nonstructural Proteins

Associated data

  • GENBANK/L37661
  • GENBANK/U94597