Research Description

Molecular Diagnosis of Dried Blood Spots from the Horn of Africa and Yemen for HIV

My project is a molecular diagnosis of dried blood spots from both the Horn of Africa and Yemen for HIV-1. The first stage of the project is assessing the quality of the blood spots. The next part is determining how many of the samples are HIV-1 positive. Finally, a rate of infection will be determined, and HIV-1 positive samples will be sequenced for HIV variation across the sample population. There are approximately 93 Horn of Africa and 550 Yemen dried blood spot samples available in my laboratory for HIV testing. After purifying the DNA, nested Polymerase Chain Reaction (PCR) is employed to amplify a segment of the HIV genome to test for the presence of the virus. My plan is to then sequence the envelope gene of those samples found to be positive to investigate the genetic variability of the virus in the population. I am also testing the Yemen blood spots for the presence of HIV-1 as preliminary data for an NIH grant that Dr. Mulligan, my mentor, is submitting. I plan to test all 550 samples, as having a large sample size will increase the power of our ability to determine HIV-1 prevalence in this country. These samples were collected from diverse ethnic and geographic areas across the entire country, and therefore I will be able to establish a rate of infection in this region. Furthermore, I am conducting extensive experiments to determine the specificity of the assay, which will establish the reliability of my results. More specifically, these tests will allow those samples that did not amplify to be either definitely HIV-1 negative or contain fewer than a determined number of copies of the viral genome.