Laboratory of Hematological Research of the Triângulo Mineiro Federal University and Uberaba Regional Blood Center - Hemominas Fundation, Uberaba, Minas Gerais, Brazil.
Laboratory of Immunology, Triângulo Mineiro Federal University, Uberaba, Minas Gerais, Brazil.
Parasitology. 2021 Jan;148(1):110-114. doi: 10.1017/S0031182020001936. Epub 2020 Nov 4.
Visceral leishmaniasis is an endemic protozoonosis observed in over 60 countries, with over 500 000 new cases recorded annually. Although the diagnostic procedure of its symptomatic forms is well established, for asymptomatic patients, who represent about 85% of those infected, there is no consensus on the best method for its identification. Recent studies have presented molecular techniques as viable identification methods, with good sensitivity and specificity indices in asymptomatic individuals. Therefore, we aimed to use molecular methods to assess their effectiveness in identifying the presence of asymptomatic infection by Leishmania infantum (L. infantum) individuals from endemic regions of Brazil. Screening was performed by real-time polymerase chain reaction (qPCR) and confirmed by sequencing the cytochrome B gene. Of the 127 samples [from 608 blood donors who had participated in a previous study, of which 34 were positive by the enzyme-linked immunosorbent assay (ELISA) rK39] tested by qPCR, 31 (24.4%) were positive. In the sequencing of 10 qPCR-positive samples, five were identified as L. infantum. Complimentary samples of the ELISA rK39 and conventional PCR showed only reasonable and low agreement with qPCR, respectively. The qPCR confirmed the presence of infection in five of the 10 sequenced samples, ELISA confirmed three, and the conventional PCR confirmed none.
内脏利什曼病是一种在超过 60 个国家流行的原生动物病,每年记录的新发病例超过 50 万例。虽然其症状形式的诊断程序已经确立,但对于无症状患者(约占感染者的 85%),其最佳识别方法尚无共识。最近的研究提出了分子技术作为可行的识别方法,在无症状个体中具有良好的敏感性和特异性指数。因此,我们旨在使用分子方法评估其在识别巴西流行地区利什曼原虫(L. infantum)无症状感染存在方面的有效性。通过实时聚合酶链反应(qPCR)进行筛查,并通过测序细胞色素 B 基因进行确认。在通过 qPCR 检测的 127 个样本[来自之前一项研究的 608 名献血者,其中 34 名通过酶联免疫吸附试验(ELISA)rK39 呈阳性]中,有 31 个(24.4%)呈阳性。在对 10 个 qPCR 阳性样本的测序中,有 5 个被鉴定为 L. infantum。ELISA rK39 和常规 PCR 的补充样本与 qPCR 的一致性分别为合理和低。qPCR 确认了 5 个测序样本中的感染,ELISA 确认了 3 个,常规 PCR 则未确认。