Diagnosis of clinical samples spotted on FTA cards using PCR-based methods.

摘要: The broad clinical presentation of Leishmaniasis makes the diagnosis current and past cases this disease rather difficult. Differential is important because diseases caused by other aetiologies a spectrum similar to that leishmaniasis (e.g. leprosy, skin cancers tuberculosis for CL; malaria schistosomiasis VL) are often present in endemic areas endemicity. Presently, variety methods have been developed tested aid identification Leishmania. advent PCR technology has opened new channels materials. simple, rapid procedure adapted leishmaniasis. A range tools currently available Leishmania species, respectively. However, none these diagnostic examined using samples spotted on FTA cards. Three different PCR-based approaches were including: kDNA minicircle, 18S rRNA gene PCR-RFLP Intergenic region ribosomal protein. primers designed sit within coding sequences genes (relatively well conserved) but which amplify across intervening intergenic sequence variable). These used reference isolates 10 most species: L. donovani, infantum, major & tropica. Digestion products with restriction enzymes produced species-specific patterns allowed discrimination isolates. minicircle highly sensitive both bone marrow smears from conserved smear less diagnosing smears. nested P5 P6 as P1 P2 newly showed high level reproducibility sensitivity. Though, it was than primers, easily discriminated between species.