Current detection schemes for Leishmania are inadequate because of lack of sensitive methods. Our goals to develop methodology for detection of sub- species of parasite in order to prescribe effective treatment regimen and to monitor blood products. Parasite DNA is an excellent tool to develop sensitive detection assays. kDNA is unique to trypanosomatids and we have taken the advantage of kDNA to develop PCR-based diagnostic strategy. We have cloned several kDNAs and determined their sequences from various isolates of Leishmania, identified conserved sequence elements and designed oligonucleotide primes to be used in PCR reaction. We are planning to develop PCR-linked, hybridization diagnostic reagents in the framework of an ELISA assay. Along with this assay we are developing Restriction Length Polymorphism (RELP) technique to analyze genomic differences between related organisms so as to distinguish sub-species of Leishmania for field clinics and to monitor blood products.
