A simplified non radioactive DNA probe technique for the field detection of sibling species A of the Anopheles culicifacies (Diptera: Culicidae) complex

Authors

  • B.G.D.N.K. De Silva Department of Zoology, University of Sri Jayewardenepura, Nugegoda, Sri Lanka
  • G. P. G. M. D. Hapugoda Department of Molecular Medicine, Faculty of Medicine,University of Kelaniya, Kelaniya, Sri Lanka
  • E. H. Karuuanayake Institute of Biochemistry, Molecular Biology and Biotechnology,of Colombo, Colombo 7, Sri Lanka

DOI:

https://doi.org/10.31357/vjs.v15i0.280

Abstract

Three cloned highly repetitive DNA sequences Rp36, Rp217 and Rp234 isolated from An.culicifacies Giles, sensu lato were developed as non radioactive DNA probes by using a biotinylatedlabeling and colorimetric detection system. These non radioactive DNA probes distinguish siblingspecies A from species B and C of the An. culicifacies complex in a dot blot hybridization assayusing single mosquito DNA extracts diluted 50 fold. The biotinylated Rp217 probe was furtherassayed in a more simple procedure which involves the hybridization of blots prepared fromsquashed mosquito heads. This technique avoids the separate extraction of mosquito DNA andfacilitates a number of samples to be processed rapidly while also allowing several field analyses tobe carried out on one mosquito specimen.

Key words : Anopheles culicifacies, non radioactive DNA probes

Author Biographies

B.G.D.N.K. De Silva, Department of Zoology, University of Sri Jayewardenepura, Nugegoda, Sri Lanka

Department of Zoology, University of Sri Jayewardenepura, Nugegoda, Sri Lanka

G. P. G. M. D. Hapugoda, Department of Molecular Medicine, Faculty of Medicine,University of Kelaniya, Kelaniya, Sri Lanka

Department of Molecular Medicine, Faculty of Medicine,University of Kelaniya, Kelaniya, Sri Lanka

E. H. Karuuanayake, Institute of Biochemistry, Molecular Biology and Biotechnology,of Colombo, Colombo 7, Sri Lanka

Institute of Biochemistry, Molecular Biology and Biotechnology,of Colombo, Colombo 7, Sri Lanka

Published

2012-02-16

Issue

Vol. 15 (2010)

Section

Articles