Rapid detection of chikungunya virus in laboratory infected Aedes aegypti by Reverse-Transcriptase- Polymerase Chain Reaction (RT-PCR)

Trop Biomed. 2005 Dec;22(2):149-54.

Abstract

A study of chikungunya virus was carried out to establish Reverse Transcriptase- Polymerase Chain Reaction (RT-PCR) as a rapid detection technique of the virus. The susceptibility of lab-colonized Aedes aegypti to chikungunya virus was also determined. Artificial membrane feeding technique was used to orally feed the mosquitoes with a human isolate of chikungunya virus. A total of 100 fully engorged female Ae. aegypti were obtained and maintained for 7 days. Seventy of them survived and then pooled at 10 individuals per pool. Total RNA was extracted from the samples and RT-PCR amplifications were carried out. Five out of 7 pools showed positive PCR band at 350-bp, indicating Ae. aegypti is a potential vector of chikungunya virus. The minimum infection rate (MIR) was 71% within these laboratory colonies. RT-PCR is a sensitive technique that is useful in detecting infected mosquitoes in epidemic areas. This technique can de used as a rapid detection method and provide an early virologic surveillance systems of chikungunya virus infected mosquitoes.

Publication types

  • Evaluation Study

MeSH terms

  • Aedes / virology*
  • Animals
  • Cell Line
  • Chikungunya virus / genetics
  • Chikungunya virus / isolation & purification*
  • Electrophoresis, Agar Gel / methods
  • Female
  • Humans
  • Insect Vectors / virology*
  • Laboratories
  • RNA, Viral* / analysis
  • RNA, Viral* / genetics
  • RNA, Viral* / isolation & purification
  • Reverse Transcriptase Polymerase Chain Reaction / methods*
  • Time Factors

Substances

  • RNA, Viral