Instituto René Rachou, Fundação Oswaldo Cruz, Grupo de Pesquisa Clínica e Políticas Públicas em Doenças Infecciosas e Parasitárias, CEP: 30190-002, Belo Horizonte, Minas Gerais, Brazil.
PLoS One. 2024 Aug 22;19(8):e0306967. doi: 10.1371/journal.pone.0306967. eCollection 2024.
Cutaneous leishmaniasis (CL) is a global public health problem caused by species on the genus Leishmania and is the most prevalent clinical form of leishmaniasis. The aim of this study was to develop a new LAMP assay for Leishmania sp. based on HSP70 gene and evaluate it clinically for molecular diagnosis of CL. The study was carried out in the following stages: i) design of primers based on HSP70 gene of Leishmania sp.; ii) evaluation of detection limit and analytical specificity; iii) estimation of the accuracy of LAMP-Leish/HSP70 assay for diagnosing CL. A total of 100 skin biopsy samples from patients, comprising 60 CL cases and 40 non-cases, were analyzed in this study. One LAMP assay using HSP70 gene as molecular target were standardized, and the observed detection limit was 100fg of L. braziliensis purified DNA. The LAMP-Leish/HSP70 assay was specific for Leishmania spp. The LAMP-Leish/HSP70 assay showed an accuracy of 92%, and positivity rates were not affected by lesion onset time or parasite load. This novel LAMP assay targeting the HSP70 gene of Leishmania sp. has the potential to be a useful tool to integrate into routine diagnosis for suspected cases of CL.
皮肤利什曼病（CL）是一种由利什曼属物种引起的全球性公共卫生问题，是利什曼病最常见的临床形式。本研究旨在基于 HSP70 基因开发一种用于 Leishmania sp. 的新型 LAMP 检测方法，并评估其在 CL 分子诊断中的临床应用。该研究分以下阶段进行：i）基于 Leishmania sp. 的 HSP70 基因设计引物；ii）评估检测限和分析特异性；iii）评估 LAMP-Leish/HSP70 检测方法诊断 CL 的准确性。本研究共分析了 100 例来自患者的皮肤活检样本，包括 60 例 CL 病例和 40 例非病例。标准化了一种基于 HSP70 基因作为分子靶标的 LAMP 检测方法，观察到的检测限为 100fg 纯化的 L. braziliensis DNA。LAMP-Leish/HSP70 检测方法对 Leishmania spp. 具有特异性。LAMP-Leish/HSP70 检测方法的准确性为 92%，并且阳性率不受病变起始时间或寄生虫负荷的影响。这种针对 Leishmania sp. HSP70 基因的新型 LAMP 检测方法具有成为疑似 CL 常规诊断有用工具的潜力。
