Subdirección General de Sistemas Terrestres, Instituto Nacional de Técnica Aeroespacial (INTA)-Campus La Marañosa, Madrid, Spain.
Faculty of Health and Medical Sciences, School of Veterinary Medicine, University of Surrey, Guildford, UK.
Transbound Emerg Dis. 2019 Nov;66(6):2474-2481. doi: 10.1111/tbed.13305. Epub 2019 Aug 8.
In this study, we describe the pathology of Leishmania infantum infection in naturally infected wild Leporidae and compare diagnosis of infection using histopathology, direct fluorescent antibody (DFA) assay, immunofluorescence antibody test (IFAT) and quantitative real-time PCR (qPCR). Tissues were analysed from 52 European rabbits (Oryctolagus cuniculus) and 7 Iberian hares (Lepus granatensis) from the Community of Madrid (Spain). Our results show that L. infantum infection is associated with only minimal histopathological lesions and that L. infantum amastigotes can be detected by DFA assay in all tissues types tested, including skin. These results were confirmed by qPCR on fresh frozen tissues in 13% of rabbits and 100% of hares. However, L. infantum DNA could not be detected by qPCR on paraffin-embedded tissue obtained by laser capture microdissection. Using the DFA assay to diagnose L. infantum, infection may provide further insights into this disease in wild animals and may allow the precise tissue localization of L. infantum, thereby guiding follow-up tests with more accurate qPCR.
在这项研究中，我们描述了天然感染的野生兔科动物内脏利什曼原虫感染的病理学，并比较了使用组织病理学、直接荧光抗体（DFA）检测、免疫荧光抗体试验（IFAT）和实时定量 PCR（qPCR）诊断感染的方法。对来自马德里社区（西班牙）的 52 只欧洲兔（Oryctolagus cuniculus）和 7 只伊比利亚野兔（Lepus granatensis）的组织进行了分析。我们的结果表明，内脏利什曼原虫感染仅与最小的组织病理学病变有关，DFA 检测可在所有检测的组织类型（包括皮肤）中检测到内脏利什曼原虫无鞭毛体。qPCR 在 13%的兔子和 100%的野兔的新鲜冷冻组织中证实了这一结果。然而，qPCR 无法在通过激光捕获显微切割获得的石蜡包埋组织中检测到内脏利什曼原虫 DNA。使用 DFA 检测诊断内脏利什曼原虫感染可能会进一步了解野生动物中的这种疾病，并可能允许对内脏利什曼原虫进行更精确的组织定位，从而指导更准确的 qPCR 进行后续检测。
