Development of DNA Probe Test for Leishmania

Our research will develop reagents and procedures for detection of nucleic acids fromLeishmania. Leishmania are grouped broadly into the New and Old World species. Throughout tropicaland subtropical regions of the world, leishmaniasis is a serious public health concern. Effectivetreatments exist; yet, diagnosis is difficult, owing to low abundance of parasites in the peripheral blood.Consensus probes and primers will be designed, based upon published mini circle kinetoplast sequences.Primers will incorporate inosines and mixed base positions to support amplification of Old and New Worldspecies. Non-radioactive microliter plate assays have been developed for performing hybridizationanalysis of amplified DNA. These methods, together with simplified DNA extraction and amplificationtechniques will be adapted to a consensus DNA probe assay for Leishmania using laboratory strains.In Phase II, these procedures will be refined and extended to detection in clinical specimens.