Department of Parasitology, Faculty of Science, Charles University, Prague, Czech Republic.
Coris BioConcept, Crealys Park, Gembloux, Belgium.
PLoS Negl Trop Dis. 2018 Aug 2;12(8):e0006607. doi: 10.1371/journal.pntd.0006607. eCollection 2018 Aug.
Canine leishmaniasis (CanL) is a zoonotic disease, caused by Leishmania infantum and transmitted by Phlebotomus perniciosus in the Mediterranean basin. Previously, an ELISA based on the P. perniciosus salivary protein SP03B was proposed as a valid tool to screen for canine exposure to sand fly bites across regions endemic for CanL. Although this approach is useful in laboratory settings, a practical tool for immediate application in the field is needed. In this study we propose the rSP03B sero-strip, the first immunochromatographic test (ICT) in the field of vector exposure able to rapidly screen dogs living in endemic areas for the presence of P. perniciosus and to aid in the evaluation of vector control programs.
METHODOLOGY/PRINCIPAL FINDINGS: The ICT was prepared using the bacterially expressed recombinant protein rSP03B as antigen. For test optimization, pre-immune sera from non-bitten laboratory-bred Beagles were used as negative controls. In order to validate the test, sera from laboratory-bred Beagles experimentally exposed to P. perniciosus bites were used as positive controls. Additionally, all samples were tested by ELISA using whole salivary gland homogenate (SGH) and the rSP03B protein as antigen. An almost perfect degree of agreement was found between the ICT and the SGH-ELISA. Furthermore, the newly proposed rSP03B sero-strip showed a sensitivity of 100% and a specificity of 86.79%.
CONCLUSIONS/SIGNIFICANCE: We developed a simple and rapid ICT based on the P. perniciosus rSP03B salivary protein, able to replace the standard ELISA used in previous studies. Our rSP03B sero-strip showed to be highly sensitive and specific in the detection of antibodies (IgG) against P. perniciosus saliva. In the future, this test can be employed during large-scale epidemiological studies of CanL in the Mediterranean area to evaluate the efficacy of vector control programs.
犬利什曼病（CanL）是一种人畜共患病，由利什曼原虫引起，通过在地中海地区的白蛉传播。此前，一种基于白蛉唾液蛋白 SP03B 的 ELISA 被提议作为一种有效的工具，用于筛选犬类是否接触过白蛉叮咬，从而检测利什曼病的流行地区。尽管这种方法在实验室环境中很有用，但需要一种实用的工具，以便在现场立即应用。在这项研究中，我们提出了 rSP03B 血清条，这是第一个能够快速筛选生活在流行地区的犬类是否存在白蛉的免疫层析检测（ICT），并有助于评估媒介控制计划。
方法/主要发现：该 ICT 采用细菌表达的重组蛋白 rSP03B 作为抗原制备。为了优化测试，使用未被白蛉叮咬的实验室饲养比格犬的预免疫血清作为阴性对照。为了验证该测试，使用实验室饲养的比格犬进行了白蛉叮咬的实验暴露，作为阳性对照。此外，所有样本均采用全唾液腺匀浆（SGH）和 rSP03B 蛋白作为抗原的 ELISA 进行检测。ICT 与 SGH-ELISA 之间存在近乎完美的一致性。此外，新提出的 rSP03B 血清条显示出 100%的敏感性和 86.79%的特异性。
结论/意义：我们开发了一种基于白蛉 rSP03B 唾液蛋白的简单快速的 ICT，能够替代以前研究中使用的标准 ELISA。我们的 rSP03B 血清条在检测针对白蛉唾液的抗体（IgG）方面表现出高度的敏感性和特异性。在未来，这种测试可以在地中海地区犬利什曼病的大规模流行病学研究中使用，以评估媒介控制计划的效果。
