Efficiency of rLb6H recombinant protein from Leishmania (Viannia) braziliensis for the detection of canine visceral leishmaniasis

Ingrid Estevam Pereira, Erick Esteves Oliveira, Carolina Martins Moreira Elias, Alexandre Barbosa Reis, Malcolm Scott Duthie, Henrique C. Teixeira
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Abstract

Introduction: Visceral leishmaniasis (VL) is a serious endemic disease in many tropical and subtropical countries, with a strong incidence in Brazil. The disease is transmitted by the bite of infected female phlebotomine sandflies, with dogs being the main urban reservoirs of the parasite. The diverse clinical profile and the long incubation period are challenges for the diagnosis of canine visceral leishmaniasis (CVL). Recombinant proteins from Leishmania spp. have been studied as antigens that can increase the accuracy of serological tests. Objective: To evaluate the diagnostic performance of the recombinant protein rLb6H, from Leishmania braziliensis, in comparison to the reference antigens rK39 and rK28, from L. donovani, prioritizing the identification of subclinical infected dogs. Material and Methods: Serum IgG reactivity to rLb6H, rK28, and rK39 recombinant proteins was assessed in dogs with previously parasitological confirmation of CVL, subdivided according to their clinical status, using immunoenzymatic assay (ELISA). Diagnostic accuracy of each ELISA was evaluated by receiver operating characteristic (ROC) curve analysis. Results: While all antigens showed a better performance in detecting CVL in symptomatic dogs (SD), detection of CVL in the oligosymptomatic (OD) and asymptomatic (AD) groups was lower, but rLb6H achieved high sensitivity for asymptomatic CVL. Interestingly, the most reactive CVL samples to rK28 were barely detected by rLb6H, while the less reactive to rK28, mostly from the AD group, presented higher reactivity to rLb6H. Conclusion: The recombinant protein rLb6H showed utility in the detection of asymptomatic CVL, displaying a complementary reactivity to rK39 and rK28. Thus, these results suggest that rLb6H could be incorporated into multi-antigen strategies, to increase diagnostic accuracy of CVL.
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巴西利什曼病(Viannia)rLb6H 重组蛋白检测犬内脏利什曼病的效率
导言:内脏利什曼病(VL)是许多热带和亚热带国家的一种严重地方病,在巴西的发病率很高。该病通过受感染的雌性血吸虫沙蝇叮咬传播,而狗是寄生虫的主要城市贮藏地。犬内脏利什曼病(CVL)的临床表现多种多样,潜伏期较长,这给诊断工作带来了挑战。研究人员已将利什曼原虫的重组蛋白作为抗原,以提高血清学检测的准确性。研究目的评估来自巴西利什曼原虫的重组蛋白 rLb6H 与来自多诺万利什曼原虫的参考抗原 rK39 和 rK28 相比的诊断性能,优先识别亚临床感染的狗。材料与方法使用免疫酶联免疫吸附试验(ELISA),对之前经寄生虫学确诊为 CVL 的犬只血清 IgG 与 rLb6H、rK28 和 rK39 重组蛋白的反应性进行评估。通过接收器操作特征曲线(ROC)分析评估了每种 ELISA 的诊断准确性。结果显示虽然所有抗原在检测有症状犬(SD)的CVL方面都有较好的表现,但在少症状犬(OD)和无症状犬(AD)组中,CVL的检测率较低,但rLb6H对无症状CVL具有较高的灵敏度。有趣的是,rLb6H 几乎检测不到对 rK28 反应性最强的 CVL 样品,而对 rK28 反应性较低的 CVL 样品(大部分来自 AD 组)对 rLb6H 的反应性较高。结论重组蛋白 rLb6H 在无症状 CVL 的检测中显示出实用性,与 rK39 和 rK28 具有互补反应性。因此,这些结果表明,可将 rLb6H 纳入多抗原策略,以提高 CVL 诊断的准确性。
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