Genetic Analysis of Aedes aegypti using Random Amplified Polymorphic DNA (RAPD) Markers from Dengue Outbreaks in Pakistan

Abstract

Background: Keeping in view the havoc situation of dengue fever in Pakistan, the current study was designed to demon­strate the genetic variations, gene flow and rate of migration from Lahore and Faisalabad. Methods: The larvae were collected from both natural and artificial breeding places from each collection site. The adult mosquitoes were collected by means of sweep net and battery-operated aspirator. DNA extraction was per­formed using TNE buffer method. Ten GeneLink-A series RAPD primers were used for PCR amplification and the data was analyzed through POPGENE. Results: The number of amplification products produced per primer varied from 8-12, ranging from 200 to 2000 bp with an average of 10.0 bands per primer. The percentage of polymorphic loci amplified by each primer varied from 22.5 to 51%. The UPGMA dendrogram demonstrates two distinct groups from Faisalabad and Lahore populations. The genetic diversity ranged from 0.260 in Faisalabad to 0.294 in Lahore with a total heterozygosity of 0.379. The GST value for nine populations within Lahore was 0.131 (Nm= 3.317), whereas for nine populations in Faisalabad GST value was 0.117 (Nm= 3.773). The overall genetic variation among eighteen populations showed GST= 0.341 and Nm= 1.966. Conclusion: The genetic relatedness and Nm value show that Ae. aegypti populations exhibit intra-population gene flow both in Faisalabad and Lahore. Although, both cities show a distinct pattern of genetic structure; however, few areas from both the cities show genetic similarity. The gene flow and the genetic relatedness in few populations of Lahore and Faisalabad cities need further investigation.

Keywords

• Aedes aegypti
• Dengue
• Genetic diversity
• RAPD markers