Implication of Random Amplified Polymorphic DNA method for differentiating Anopheles culicifacies sibling species
Varun Tyagi, Ajay K. Sharma, Ruchi Yadav, T. Adak, D. Sukumaran, O. P. Agrawal, Vijay Veer
Efficient method for discriminating among members of Anopheline species complexes is of extremernsignificance for epidemiological studies and effective vector control programme. Anopheles culicifaciesrnis a principal malaria vector in rural, periurban and tribal settings and responsible for about 65- 70%rnmalaria cases in India. It is a complex of 5 isomorphic types A, B, C, D and E with varying biologicalrncharacteristics. Presently, cytogenetic methods involving polytene chromosome and mitoticrnkaryotyping are the only methods that could differentiate all the five sibling species but theserntechniques are time consuming and labour intensive. DNA based techniques using rDNA and COIIrnsuch as AS-PCR and gene specific PCR-RFLP seems to be good alternatives but these techniques toornhave limitations and until now none of the techniques have been able to differentiate all the five siblingrnspecies in one step. In the present study we have been able to develop Random Amplified PolymorphicrnDNA (RAPD) markers that could differentiate all the five sibling species of An. culicifacies in India. Arnnumber of RAPD primers were screened and genetic fingerprints for all the 5 sibling species of An.rnculicifacies were developed using a selected primer. A total of 34 loci representing these sibling speciesrnusing selected primer were polymorphic and able to differentiate the sibling species and showed 100%rnpolymorphism. A consensus tree was also generated indicating genetic distances among these siblingrnspecies. The findings will be useful in devising a new method for identification of sibling species whichrnwill be helpful to develop successful vector management strategies.
Varun Tyagi, Ajay K. Sharma, Ruchi Yadav, T. Adak, D. Sukumaran, O. P. Agrawal, Vijay Veer. Implication of Random Amplified Polymorphic DNA method for differentiating Anopheles culicifacies sibling species. European Journal of Biotechnology and Bioscience, Volume 3, Issue 8, 2015, Pages 47-54