Programa de Pós-Graduação em Biologia Molecular e Celular Aplicada à Saúde, Universidade Luterana do Brasil (ULBRA), Canoas, RS, Brazil.
Programa de Pós-Graduação em Patologia, Universidade Federal de Ciências da Saúde de Porto Alegre (UFCSPA), Porto Alegre, Rio Grande do Sul, 90050-170, Brazil.
Epidemiol Infect. 2018 Apr;146(5):571-576. doi: 10.1017/S0950268818000225. Epub 2018 Feb 26.
Although serological assays have been widely used for the diagnosis of canine visceral leishmaniasis (CVL), they present different performances depending on the clinical profile of the dogs. This study evaluated the accuracy of serological tests, immunochromatographic (Dual Path Platform: DPP®) and enzyme-linked immunosorbent (ELISA EIE®), for CVL in relation to the detection of Leishmania DNA through real-time polymerase chain reaction (real-time PCR) in samples from symptomatic and asymptomatic dogs from a non-endemic area in the state of Rio Grande do Sul, Southern Brazil. Serum from 140 dogs (39 symptomatic and 101 asymptomatic) was tested by DPP and ELISA followed by real-time PCR. From a total of 140 samples evaluated, Leishmania DNA was detected by real-time PCR in 41.4% (58/140). Moreover, 67.2% of samples positive in real-time PCR were positive in both DPP and ELISA (39/58), showing moderate agreement between methods. In the symptomatic group, one sample non-reactive in both serological assays was positive in real-time PCR, whereas in the asymptomatic group, 17.8% non-reactive or undetermined samples in serological assays were positive in the molecular method. Leishmania DNA was not detected in 17.9% reactive samples by serological assays from the symptomatic group, and in 3.9% from asymptomatic dogs. Real-time PCR demonstrated greater homogeneity between symptomatic and asymptomatic groups compared with DPP and ELISA. The molecular method can help to establish the correct CVL diagnosis, particularly in asymptomatic dogs, avoiding undesirable euthanasia.
尽管血清学检测已广泛用于犬内脏利什曼病(CVL)的诊断,但它们的表现因犬的临床特征而异。本研究评估了血清学检测(免疫层析法[双路径平台:DPP®]和酶联免疫吸附法[EIE®])在巴西南里奥格兰德州非流行地区的症状性和无症状性犬中针对 CVL 的准确性,同时检测了通过实时聚合酶链反应(real-time PCR)从这些犬中采集的样本中的利什曼原虫 DNA。使用 DPP 和 ELISA 对来自 140 只犬(39 只症状性和 101 只无症状性)的血清进行了检测,然后进行 real-time PCR。在总共评估的 140 个样本中,通过 real-time PCR 检测到 41.4%(58/140)的样本呈阳性。此外,在 real-time PCR 中呈阳性的样本中,有 67.2%(39/58)在 DPP 和 ELISA 中也呈阳性,表明两种方法之间具有中等一致性。在症状性组中,1 个在两种血清学检测中均呈阴性的样本在 real-time PCR 中呈阳性,而在无症状组中,17.8%的血清学检测呈阴性或不确定的样本在分子方法中呈阳性。在症状性组中,有 17.9%的反应性样本在血清学检测中未检测到利什曼原虫 DNA,而在无症状组中,有 3.9%的样本未检测到。与 DPP 和 ELISA 相比,real-time PCR 显示症状性组和无症状性组之间的一致性更高。该分子方法有助于确立正确的 CVL 诊断,特别是在无症状性犬中,避免不必要的安乐死。