Identification of leishmania by kinetoplast DNA minicircle and cloning of minicircle.
To develop a specific and sensitive method for the identification of Leishmania species and strains in China, Kinetoplast DNA (kDNAs) from 10 different isolates of Leishmania donovani and other species were analyzed by Southern and Dot hybridization. Our results showed significant diversities within different species and strains of Leishmania, which may be a useful criterion for taxonomic analysis. By Southern blot under different stringency conditions, we estimated that there is greater sequence heterogeneity between L. donovani isolated from dog and from human being in the same endemic focus than within the same isolate. Our Dot blotting assay with kDNA probe labelled by photobiotin differentiated species and isolates of Leishmania with as little as 10(4) parasites; so it is useful for field identification. The heterogeneity between isolates of L. donovani from patients in hilly and plain areas may have practical implications for disclosing the difference in endemicity. For the purpose of obtaining a large quantity of homogeneous minicircle, it was cloned and produced by recombinant pUC18.
Lu
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HG
Laboratory of Parasitology, West-China University of Medical Sciences, Chengdu.
Hu
X S
XS
D000818:Animals; D015139:Blotting, Southern; D003001:Cloning, Molecular; D015342:DNA Probes; D004270:DNA, Circular / Q000032:analysis*; D018105:DNA, Kinetoplast; D006801:Humans; D007893:Leishmania donovani / Q000145:classification* / Q000302:isolation & purification
D015342:DNA Probes; D004270:DNA, Circular; D018105:DNA, Kinetoplast