Leishmania sp. identification by PCR associatedwith sequencing of target SSU rDNA in paraffin-embeddedskin samples stored for more than 30 years
Lima, Ana Carolina S. de; Zampieri, Ricardo A; Tomokane, Thaíse Y; Laurenti, Márcia D; Silveira, Fernando T; Corbett, Carlos E. P; Floeter-Winter, Lucile M; Gomes, Cláudia M. C.
Parasitol. res
; 108: 1525-1531, 2011. tab, ilus, graf
Artículo
en Inglés
| BVSDIP, FIOCRUZ | ID: dip-3122
Paraffin-embedded samples commonly stored ateducational and research institutions constitute tissues banksfor follow-up or epidemiological studies; however, theparaffin inclusion process involves the use of substances thatcan cause DNA degradation. In this study, a PCR protocolwasapplied to identify Leishmania strains in 33 paraffinembeddedskin samples of patients with American cutaneousleishmaniasis. DNA was obtained by the phenol-chloroformprotocol following paraffin removal and then used in PCR ornested PCR based on the nucleotide sequence of the smallsubunit ribosomal RNA (SSU rDNA). The ampliconsobtained were cloned and sequenced to determine the singlenucleotide polymorphism that distinguishes between differentLeishmania species or groups. This assay allowed todistinguish organisms belonging to the subgenus Viannia andidentify L. (Leishmania) amazonensis and L. (L.) chagasi ofthe Leishmania subgenus. Of the 33 samples, PCR andnested PCR identified 91 percent of samples. After sequencing thePCR product of 26 samples, 16 were identified as L. (L.) (AU)
Biblioteca responsable:
BR275.1
Ubicación: BR275.1; PCIEC2011
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