Departament de Medicina i Cirurgia Animals, Facultat de Veterinària, Universitat Autònoma de Barcelona, Cerdanyola del Vallès, Barcelona, Spain.
Departament de Biologia, Sanitat i Medi Ambient, Facultat de Farmàcia i Ciències de l'Alimentació, Universitat de Barcelona, Barcelona, Barcelona, Spain.
Parasit Vectors. 2022 May 10;15(1):164. doi: 10.1186/s13071-022-05246-2.
Canine leishmaniosis caused by the protozoan Leishmania infantum is a complex infection due to its variable clinical signs and laboratory findings. Therefore, a broad range of techniques is available for diagnosis. Testing for specific antibodies in serum is the most commonly used technique, although the testing of other body fluids, such as oral transudate (OT), can be an alternative as its collection is non-invasive and testing can be performed by untrained personnel. The aim of this study was to assess and compare the detection of L. infantum-specific antibodies in paired samples of serum and OT collected from apparently healthy dogs and dogs with clinical leishmaniosis using an in-house enyzme-linked immunosorbent assay (ELISA).
Serum and OT were collected from 407 dogs, which varied in breed, sex, age, lifestyle and clinical status, by many practicing veterinarians in Spain. The main geographical areas of sampling included Barcelona (n = 110), Mallorca (n = 94), Cadiz (n = 54) and Asturias (n = 47). The majority of infected dogs were apparently healthy (89.9%) while 41 presented clinical signs and/or clinicopathological abnormalities compatible with L. infantum infection and subsequently diagnosed with leishmaniosis (10.1%). An in-house ELISA was performed to quantify the anti-Leishmania antibodies in serum and OT.
The L. infantum infection rate determined by the in-house ELISA was 37.1% in serum samples and 32.7% in OT samples. Serum and OT ELISA results showed a positive correlation (Spearman's correlation coefficient r = 0.6687, P < 0.0001). The percent agreement between the serum and OT ELISA results was 84%, while agreement according to Cohen's kappa statistic (κ) was substantial (0.66) when all samples were analyzed. The highest percent agreement (92.1%) between both tests was found in dogs from low endemicity regions and from sick dogs, with both groups presenting almost perfect agreement according to Cohen's κ agreement test (0.84). Few seronegative dogs (n = 23) tested positive by the OT ELISA. The agreement between serum and OT went from almost perfect to moderate when the geographical distribution and clinical status were analyzed.
The results of this study demonstrated an almost perfect to moderate agreement between OT and serum samples tested using the in-house ELISA. These results are particularly promising in sick dogs with high antibody levels while the results seem less optimal in apparently healthy dogs with low antibody levels.
由原生动物利什曼原虫引起的犬利什曼病是一种复杂的感染,因为其临床表现和实验室结果具有多变性。因此,有多种技术可用于诊断。检测血清中的特异性抗体是最常用的技术,尽管检测其他体液,如口腔渗出液(OT),也可以作为替代方法,因为其采集是非侵入性的,并且可以由未经培训的人员进行检测。本研究旨在评估和比较使用内部酶联免疫吸附试验(ELISA)检测来自貌似健康的犬和患有临床利什曼病的犬的配对血清和 OT 样本中利什曼原虫特异性抗体的检测。
由西班牙许多执业兽医采集 407 只犬的血清和 OT,这些犬的品种、性别、年龄、生活方式和临床状况各不相同。主要采样地理区域包括巴塞罗那(n=110)、马略卡岛(n=94)、加的斯(n=54)和阿斯图里亚斯(n=47)。大多数感染犬表现为貌似健康(89.9%),而 41 只犬出现与利什曼原虫感染相符的临床症状和/或临床病理异常,随后被诊断为利什曼病(10.1%)。使用内部 ELISA 定量检测血清和 OT 中的抗利什曼原虫抗体。
内部 ELISA 检测到的利什曼原虫感染率在血清样本中为 37.1%,在 OT 样本中为 32.7%。血清和 OT ELISA 结果呈正相关(Spearman 相关系数 r=0.6687,P<0.0001)。当分析所有样本时,血清和 OT ELISA 结果的一致性为 84%,而根据 Cohen 的 κ 统计(κ)的一致性为适度(0.66)。在低流行地区和患病犬中,两种检测方法的一致性最高(92.1%),根据 Cohen 的 κ 一致性检验,这两组的一致性几乎是完美的(0.84)。少数血清阴性犬(n=23)通过 OT ELISA 检测呈阳性。当分析地理分布和临床状况时,血清和 OT 之间的一致性从几乎完美到中度。
本研究结果表明,使用内部 ELISA 检测的 OT 和血清样本之间具有几乎完美到中度的一致性。这些结果在高抗体水平的患病犬中尤其有希望,而在低抗体水平的貌似健康的犬中,结果似乎不太理想。