TGMACK, avaliação de um novo teste molecular para o diagnóstico da leishmaniose visceral
Data
2017-08-15
Autores
Silva, Amanda Evelyn da [UNESP]
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Editor
Universidade Estadual Paulista (Unesp)
Resumo
Leishmania spp. são protozoários pertencentes à família Trypanosomatidae e agentes etiológicos das leishmanioses. Estima-se que estão em risco de contrair leishmanioses 350 milhões de pessoas em 98 países em diferentes continentes. Dentre as manifestações clínicas existentes, leishmaniose cutânea, mucocutânea e visceral, esta última é a mais severa e apresenta o maior número de óbitos. A Leishmania infantum é o agente causador da leishmaniose visceral nas Américas, na China e na bacia do Mediterrâneo, apresentando o cão como reservatório de importância epidemiológica. A Leishmania é um modelo experimental biológico que vem sendo explorado por vários pesquisadores ao redor do mundo, e isso tem proporcionado estudos por meio de tecnologias genômicas, proteômicas e metabolômicas. Muitas das sequências estudadas dentro desse grupo de parasitos são conservadas e são gênero ou, até mesmo, espécie específicas e têm sido utilizadas para diagnósticos moleculares. Para esse último são requeridos testes de alta sensibilidade e especificidade. De modo a proporcionar maior sensibilidade aos testes, novas ferramentas baseadas em métodos moleculares estão sendo desenvolvidas. Dentro desse contexto, este estudo visa avaliar a aplicação de um gene específico para L. infantum, denominado TGMACK na utilização em diagnóstico molecular de leishmaniose visceral canina, por meio de técnicas de PCR (Reação em Cadeia da Polimerase) convencional e em tempo real. Este trabalho revelou que o gene em estudo é específico para a espécie L. infantum, não havendo reação cruzada com outros parasitos do mesmo gênero. Além disso, foi possível realizar a quantificação da carga parasitária com os oligonucleotídeos 18S dos cães naturalmente infectados com L. infantum e com Lv112 e correlacionar com dados clínicos e de diagnósticos sorológico, reforçando o fato que o diagnóstico molecular apresenta grande importância no conjunto de uma análise de uma doença, principalmente pelo fato de quantificar os parasitos presente nos cães, podendo prevenir a disseminação da doença. Com base nestes estudos, pode-se concluir que o gene TGMACK é um potente candidato para ser utilizado no diagnóstico de LV por PCR em tem real.
Leishmania spp. are protozoa belonging to the family Trypanosomatidae and etiological agents of leishmaniasis. It is estimated that 350 million people are at risk of contracting leishmaniasis in 98 countries on different continents. Among the existing clinical manifestations, cutaneous, mucocutaneous and visceral leishmaniasis, the latter is the most severe and presents the highest number of deaths. Leishmania infantum is the causative agent of visceral leishmaniasis in the Americas, China and the Mediterranean Basin, presents the dog as a reservoir of epidemiological importance. Leishmania is an experimental biological model that has been explored by several researchers around the world, and this is an example for the genome, proteomics and metabolomics technologies. Many of the sequences studied within the parasite group are conserved and are genus or even specific species and have been difficult for molecular diagnostics. For the latter, tests of high sensitivity and specificity are required. In order to provide more information on testing, new tools based on molecular methods are being developed. In the context, this study aims to evaluate an application of a specific gene for L. infantum, denominated TGMACK in the use in molecular diagnosis of canine / human visceral leishmaniasis, by means of PCR (Polymerase Chain Reaction) techniques. This work revealed that the gene under study is specific especies for the L. infantum, and there is no cross-reaction with other parasites of the same genus. In addition, it was possible to quantify the parasite load with 18S oligonucleotides from dogs naturally infected with L. infantum and with Lv112 and to correlate with clinical and serological diagnostic data, reinforcing the fact that molecular diagnosis has great importance in the set of analysis of a disease, mainly by the fact of quantifying the parasites present in the dogs, being able to prevent the spread of the disease. Based on these studies, it can be concluded that the TGMACK gene is a potent candidate for use in the diagnosis of VL by real-time PCR.
Leishmania spp. are protozoa belonging to the family Trypanosomatidae and etiological agents of leishmaniasis. It is estimated that 350 million people are at risk of contracting leishmaniasis in 98 countries on different continents. Among the existing clinical manifestations, cutaneous, mucocutaneous and visceral leishmaniasis, the latter is the most severe and presents the highest number of deaths. Leishmania infantum is the causative agent of visceral leishmaniasis in the Americas, China and the Mediterranean Basin, presents the dog as a reservoir of epidemiological importance. Leishmania is an experimental biological model that has been explored by several researchers around the world, and this is an example for the genome, proteomics and metabolomics technologies. Many of the sequences studied within the parasite group are conserved and are genus or even specific species and have been difficult for molecular diagnostics. For the latter, tests of high sensitivity and specificity are required. In order to provide more information on testing, new tools based on molecular methods are being developed. In the context, this study aims to evaluate an application of a specific gene for L. infantum, denominated TGMACK in the use in molecular diagnosis of canine / human visceral leishmaniasis, by means of PCR (Polymerase Chain Reaction) techniques. This work revealed that the gene under study is specific especies for the L. infantum, and there is no cross-reaction with other parasites of the same genus. In addition, it was possible to quantify the parasite load with 18S oligonucleotides from dogs naturally infected with L. infantum and with Lv112 and to correlate with clinical and serological diagnostic data, reinforcing the fact that molecular diagnosis has great importance in the set of analysis of a disease, mainly by the fact of quantifying the parasites present in the dogs, being able to prevent the spread of the disease. Based on these studies, it can be concluded that the TGMACK gene is a potent candidate for use in the diagnosis of VL by real-time PCR.
Descrição
Palavras-chave
Leishmania infantum, Diagnóstico molecular, PCR convencional, PCR em tempo real, Leishmaniose visceral