Rihs José Bryan, Vilela Mariana Teixeira, Dos Santos Janete Soares Coelho, Caldas Sérgio, Leite Rodrigo Souza, Mol Marcos Paulo Gomes
Ezequiel Dias Foundation, Research and Development Department, Rua Conde Pereira Carneiro, 80 - Gameleira, Belo Horizonte - MG, 30510-010, Brazil.
Laboratory of Immunobiology and Control of Parasites, Postgraduate Program in Parasitology, Institute of Biological Sciences, Federal University of Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
Parasitol Res. 2025 May 21;124(5):54. doi: 10.1007/s00436-025-08503-2.
Real-time PCR (qPCR) has emerged as a promising alternative to be used in the diagnosis of leishmaniasis. However, a consensus on the standardized approach for this purpose is yet to be established. This systematic review aims to highlight relevant aspects for the development of possible consensus protocols. The study included publications from January 2011 to July 2023 using different databases. Studies focusing on qPCR standardization for the diagnosis of leishmaniasis were included. The collected data encompassed study group details, geographical locations, sample types and quantities, molecular targets, and qPCR methodologies, which were subsequently pooled and analyzed. Forty-three studies conducted in diverse regions of the world were included. The majority were visceral leishmaniasis, followed by cutaneous leishmaniasis and post-kala-azar dermal leishmaniasis. Overall, invasive samples demonstrated higher sensitivity (> 90%) compared to non-invasive samples (< 90%), but both exhibited high specificity (> 85%). Furthermore, the kDNA minicircle was identified as the most promising molecular target, with a discreet preference for the use of TaqMan probes over SYBR Green. Various qPCR approaches have exhibited promising results in the diagnosis of leishmaniasis. However, further studies exploring non-invasive sampling techniques in conjunction with appropriate molecular targets are essential to establish efficient and reliable diagnostic techniques.
实时荧光定量聚合酶链反应（qPCR）已成为用于利什曼病诊断的一种有前景的替代方法。然而，针对此目的的标准化方法尚未达成共识。本系统评价旨在突出可能达成共识方案的相关方面。该研究纳入了2011年1月至2023年7月期间使用不同数据库的出版物。纳入了专注于利什曼病诊断的qPCR标准化的研究。收集的数据包括研究组详细信息、地理位置、样本类型和数量、分子靶点以及qPCR方法，随后进行汇总和分析。纳入了在世界不同地区进行的43项研究。大多数是内脏利什曼病，其次是皮肤利什曼病和黑热病后皮肤利什曼病。总体而言，侵入性样本的敏感性（>90%）高于非侵入性样本（<90%），但两者均表现出高特异性（>85%）。此外，kDNA小环被确定为最有前景的分子靶点，与SYBR Green相比，对TaqMan探针的使用有明显偏好。各种qPCR方法在利什曼病诊断中均显示出有前景的结果。然而，进一步研究探索结合适当分子靶点的非侵入性采样技术对于建立高效可靠的诊断技术至关重要。
