Pereira Diego Carlos Andrade, Teixeira-Neto Rafael Gonçalves, Lopes Valeriana Valadares, Pena Héber Paulino, Paz Gustavo Fontes, Custodio Carlos Henrique Xavier, Belo Vinícius Silva, da Fonseca Júnior Antônio Augusto, da Silva Eduardo Sérgio
Universidade Federal de São João Del Rei (UFSJ), Campus Centro-Oeste Dona Lindu, Divinópolis, Brazil.
Centro de Pesquisas René Rachou/CPqRR, Belo Horizonte, Brazil.
Mol Cell Biochem. 2023 Nov;478(11):2445-2450. doi: 10.1007/s11010-023-04672-9. Epub 2023 Feb 15.
Leishmaniasis is a zoonotic disease with worldwide distribution. In the Americas, the causative agent of the visceral form is the protozoa Leishmania (Leishmania) infantum. Transmission to the host or vertebrate reservoir occurs through the bite of infected arthropod females like Lutzomyia longipalpis. The epidemiological connection between the infection in dogs and humans generate constant studies about the relationship between the parasite and the canine host, including the development of methods and tests for the detection and quantification ofLeishmania (L.) infantum. Both conventional PCR (cPCR) and quantitative PCR (qPCR) can be used in the diagnosis of the parasite. Dropet Digital PCR (ddPCR) is another useful tool. Knowing the parasite load and its relationship with the clinical signs of naturally infected dogs is useful in research development and for establishing treatments that reduce the transmission of the disease. In this study, thirty-nine clinical samples of spleen from dogs naturaly infected by L. infantum were collected after necropsy. Two molecular tools were used to quantify the parasite load (qPCR and ddPCR) and there was 100% agreement in the results of the them. The tools developed in this work are important for the detection of L. infantum in dogs and humans. Droplet Digital PCR does not require a standard curve and is easy to standardize. In such manner, this new tool can generate more in-depth information in the broad debate about parasitic loads and the pathogenesis of leishmaniasis.
利什曼病是一种分布于全球的人畜共患病。在美洲，内脏型利什曼病的病原体是原生动物婴儿利什曼原虫（Leishmania (Leishmania) infantum）。感染通过受感染的节肢动物雌性（如长须罗蛉）叮咬传播给宿主或脊椎动物宿主。犬类和人类感染之间的流行病学联系促使人们不断研究寄生虫与犬类宿主之间的关系，包括开发检测和定量婴儿利什曼原虫（Leishmania (L.) infantum）的方法和测试。传统PCR（cPCR）和定量PCR（qPCR）均可用于该寄生虫的诊断。液滴数字PCR（ddPCR）是另一种有用的工具。了解寄生虫载量及其与自然感染犬类临床症状的关系，对研究发展以及制定减少疾病传播的治疗方法很有用。在本研究中，对39例经尸检确诊为自然感染婴儿利什曼原虫的犬类脾脏临床样本进行了分析。使用两种分子工具（qPCR和ddPCR）对寄生虫载量进行定量，结果显示二者完全一致。本研究开发的工具对检测犬类和人类中的婴儿利什曼原虫具有重要意义。液滴数字PCR不需要标准曲线，易于标准化。因此，这种新工具能够在关于寄生虫载量和利什曼病发病机制的广泛讨论中提供更深入的信息。
