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ArtigoDireito Autoral
Acesso restrito
Data de embargo
2030-01-01
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- IOC - Artigos de Periódicos [12642]
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THE APPLICABILITY OF REAL-TIME PCR IN THE DIAGNOSTIC OF CUTANEOUS LEISHMANIASIS AND PARASITE QUANTIFICATION FOR CLINICAL MANAGEMENT: CURRENT STATUS AND PERSPECTIVES
Leishmaniose cutânea
Diagnóstico
Reação em Cadeia da Polimerase em Tempo Real
Detecção
Quantificação
identificação de espécies
diagnóstico molecular
estandardização
Cutaneous leishmaniasis
diagnostic
PCR-based methods
PCR assays
real-time PCR
detection
quantification
species identification
molecular diagnosis
standardization
Afiliação
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Biologia Molecular e Doenças Endêmicas. Rio de Janeiro, RJ. Brasil.
World Health Organization. Pan-American Health Organization. Rio de Janeiro, RJ, Brasil
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisas em Leishmanioses. Rio de Janeiro, RJ. Brasil.
World Health Organization. Pan-American Health Organization. Rio de Janeiro, RJ, Brasil
Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Pesquisas em Leishmanioses. Rio de Janeiro, RJ. Brasil.
Resumo em Inglês
Cutaneous leishmaniasis (CL) is spread worldwide and is the most common manifestation of leishmaniasis. Diagnosis is performed by combining clinical and epidemiological features, and through the detection of Leishmania parasites (or DNA) in tissue specimens or trough parasite isolation in culture medium. Diagnosis of CL is challenging, reflecting the pleomorphic clinical manifestations of this disease. Skin lesions vary in severity, clinical appearance, and duration, and in some cases, they can be indistinguishable from lesions related to other diseases. Over the past few decades, PCR-based methods, including real-time PCR assays, have been developed for Leishmania detection, quantification and species identification, improving the molecular diagnosis of CL. This review provides an overview of many real-time PCR methods reported for the diagnostic evaluation of CL and some recommendations for the application of these methods for quantification purposes for clinical management and epidemiological studies. Furthermore, the use of real-time PCR for Leishmania species identification is also presented. The advantages of real-time PCR protocols are numerous, including increased sensitivity and specificity and simpler standardization of diagnostic procedures. However, despite the numerous assays described, there is still no consensus regarding the methods employed. Furthermore, the analytical and clinical validation of CL molecular diagnosis has not followed international guidelines so far. A consensus methodology comprising a DNA extraction protocol with an exogenous quality control and an internal reference to normalize parasite load is still needed. In addition, the analytical and clinical performance of any consensus methodology must be accurately assessed. This review shows that a standardization initiative is essential to guide researchers and clinical laboratories towards the achievement of a robust and reproducible methodology, which will permit further evaluation of parasite load as a surrogate marker of prognosis and monitoring of aetiological treatment, particularly in multi-centric observational studies and clinical trials.
Palavras-chave
LeishmanioseLeishmaniose cutânea
Diagnóstico
Reação em Cadeia da Polimerase em Tempo Real
Detecção
Quantificação
identificação de espécies
diagnóstico molecular
estandardização
Palavras-chave em inglês
LeishmaniasisCutaneous leishmaniasis
diagnostic
PCR-based methods
PCR assays
real-time PCR
detection
quantification
species identification
molecular diagnosis
standardization
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