About four years ago is when I realized I wanted to do something that made a difference in the area of global health and shortly after I transferred to USF, I started volunteering at the global infectious disease research lab. Dr. Thomas Unnasch, chairman of the dept. of global health, is the lab director and has multiple projects involving research in river blindness, eastern equine encephalitis, and malaria. Sadly, malaria has plagued millions around the world and can be considered a pandemic; according to WHO, the 2015 World Malaria Report calculated a prevalence rate of 214 million and mortality rate of 438,000. The project I am currently working on pertains to identifying which mosquitoes collected from Uganda are positive for carrying the Plasmodium falciparum parasite, which causes the most malignant type of malaria in humans. Since these mosquito samples come from three separate villages in Uganda, this also helps to aid in the surveillance of the disease as well as surveillance of the mosquitoes themselves within each village.
However, to obtain these negative or positive results for P. falciparum, a protocol known as Mosquito DNA Isolation Using DNAzol must be performed. The above picture shows a crushed female Anopheles mosquito that is being prepared for the addition of ethanol, a preservative that also helps to concentrate the DNA. By performing several other steps in the protocol, the end result is to hopefully have a pure DNA sample resting invisibly at the bottom of the tube without any crushed mosquito parts; left over mosquito parts can cause a problem later on when performing a test to see visible results. After DNA extraction has been completed, the next step is to perform a PCR (polymerase chain reaction) in order to amplify the specific section of DNA needed; this takes about four hours to complete so fortunately, it is all done by a small machine.
The above photo illustrates results of another process known as electrophoresis, or the movement of molecules through an agarose gel stimulated by an electric current. To get a clear picture showing multiple mosquito samples positive for Plasmodium, indicated by the bright horizontal markings present at the same level, it is important to have minimal air bubbles in the gel, no mosquito parts in DNA samples, no contamination of DNA, and a proper loading dye to DNA sample ratio; regardless of the sample size, the ratio is most often 1 parts DNA and 5 parts loading dye since that dye is what anchors down the DNA for better, more visible end results. This is the final step in identifying which mosquitoes from what village are infected with malaria and once it is complete, the results are recorded in excel.