Diferenciação molecular de subgênero, complexo e espécies de Leishmania
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Data
2018-08-01
Autores
Trigo, Beatriz Batista
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
As diferentes espécies de Leishmania causam amplo espectro de manifestações clínicas sendo genericamente classificadas em leishmaniose cutanêa (LC), mucocutânea (LMC) e leishmaniose visceral (LV), acometendo tanto crianças como adultos, e várias espécies de mamíferos. Para a identificação da espécie faz-se necessário o isolamento do protozoário a partir de amostras biológicas e o diagnóstico em laboratórios de referência. Atualmente, o “padrão ouro” para identificação de cepas de Leishmania spp. em estudos epidemiológicos é o multilocus enzyme electrophoresis (MLEE), a qual é uma técnica laboriosa e de aplicação restrita à amostras em cultivo. As técnicas baseadas na Reação em Cadeia da Polimerase (PCR) têm sido bastante utilizadas para caracterizar e diferenciar as espécies de Leishmania. Tal diferenciação pode ser importante para a determinação do tratamento correto aos pacientes, diminuindo a letalidade, e para estudos epidemiológicos com o mapeamento do padrão de distribuição das leishmanioses, auxiliando assim na priorização das medidas de controle desta zoonose. A presente pesquisa teve como objetivo avaliar in silico os oligonucleotídeos iniciadores já utilizados no diagnóstico molecular e validar, por PCR em Tempo Real, sua especificidade em cepas de referência cultivadas in vitro.
Different species of Leishmania cause a wide spectrum of clinical manifestations which are generically classified as cutaneous leishmaniasis (CL), mucocutaneous (MCL) and visceral leishmaniasis (VL), affecting children and adults, as well as several species of mammals. Species identification requires isolating the protozoan from biological samples and the use of diagnostic techniques done in reference laboratories. Currently, the "gold standard" for identifying strains of Leishmania spp. in epidemiological studies is the multilocus enzyme electrophoresis (MLEE), which is a laborious technique and restricted used in cultured samples. Techniques based on the Polymerase Chain Reaction (PCR) are being more frequently used in order to characterize and differentiate Leishmania species. Such differentiation is important to indicate the adequate treatment for the patients minimizing lethality and for epidemiological purposes as mapping its distribution pattern, in order to help prioritizing the control measures for this zoonosis. The present research aimed to evaluate the currently used primers for molecular diagnosis and validate them by Real Time PCR for their specificity in reference strains cultivated in vitro.
Different species of Leishmania cause a wide spectrum of clinical manifestations which are generically classified as cutaneous leishmaniasis (CL), mucocutaneous (MCL) and visceral leishmaniasis (VL), affecting children and adults, as well as several species of mammals. Species identification requires isolating the protozoan from biological samples and the use of diagnostic techniques done in reference laboratories. Currently, the "gold standard" for identifying strains of Leishmania spp. in epidemiological studies is the multilocus enzyme electrophoresis (MLEE), which is a laborious technique and restricted used in cultured samples. Techniques based on the Polymerase Chain Reaction (PCR) are being more frequently used in order to characterize and differentiate Leishmania species. Such differentiation is important to indicate the adequate treatment for the patients minimizing lethality and for epidemiological purposes as mapping its distribution pattern, in order to help prioritizing the control measures for this zoonosis. The present research aimed to evaluate the currently used primers for molecular diagnosis and validate them by Real Time PCR for their specificity in reference strains cultivated in vitro.
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Palavras-chave
Diagnóstico diferencial, leishmaniose, Reação em cadeia da polimerase em Tempo Real, Differential diagnosis, leishmaniasis, Real-time polymerase chain reaction