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巴西皮奥伊州无症状感染利什曼原虫个体检测方法的评估。

Evaluation of methods for detection of asymptomatic individuals infected with Leishmania infantum in the state of Piauí, Brazil.

机构信息

Department of Biochemistry and Immunology, Ribeirão Preto School of Medicine, University of São Paulo, Ribeirão Preto, SP, Brazil.

Institute of Tropical Diseases Natan Portela, Federal University of Piauí, Brazil.

出版信息

PLoS Negl Trop Dis. 2019 Jul 1;13(7):e0007493. doi: 10.1371/journal.pntd.0007493. eCollection 2019 Jul.

Abstract

BACKGROUND

Visceral Leishmaniasis in humans presents with fever, anemia, and splenomegaly and can be lethal if not treated. Nevertheless, the majority of Leishmania infantum-infected individuals does not manifest symptoms and remain so provided they are not immunosuppressed. In this work, the performance of different tests was evaluated to detect asymptomatic individuals who were living in Teresina, Piauí state, Brazil, an endemic area for VL.

METHODOLOGY

L. infantum-specific antibodies were detected by ELISA and two different rapid immunochromatographic (IC) diagnostic tests, Kalazar Detect and OnSite, and parasitic loads were detected by real time PCR [qPCR]. Additionally, we measured levels of the biomarkers monokine induced by IFN-γ (MIG) and IFN-γ-induced protein 10 (IP-10) before and after stimulation of whole blood with soluble Leishmania antigen [SLA].

PRINCIPAL FINDINGS

Kalazar Detect and OnSite detected, respectively, 76% and 64% of patients presenting with active Visceral Leishmaniasis; 50% and 57% of patients remained positive in these tests, respectively, after treatment. Of the healthy participants in the study who were living in the endemic area, only 1.7% were positive with both of the IC tests. On the other hand, reactivity in ELISA tests revealed that 13% of these individuals presented asymptomatic infections; among VL patients, 84% presenting with active disease were reactive in ELISA, and after treatment, 55.5% were seropositive. L. infantum DNA was present in the blood of 37.9% of infected individuals living in the endemic area, while IP-10 and MIG biomarkers were detected in 26.7% of them. The greatest concordance of positivity occurred between ELISA and qPCR.

CONCLUSION

The association of different techniques can detect asymptomatic infections, however, more research is necessary to develop ideal biomarkers that are simple to use in the clinic and in field studies in areas endemic for Visceral Leishmaniasis.

摘要

背景

人类内脏利什曼病的表现为发热、贫血和脾肿大,如果不治疗可能致命。然而,大多数感染利什曼原虫的人没有表现出症状,如果他们没有免疫抑制,就会保持这种状态。在这项工作中,评估了不同检测方法的性能,以检测生活在巴西皮奥伊州特雷西纳的无症状个体,该地区是内脏利什曼病的流行区。

方法

通过 ELISA 和两种不同的快速免疫层析(IC)诊断检测试剂盒,即 Kalazar Detect 和 OnSite,检测利什曼原虫特异性抗体,并通过实时 PCR [qPCR]检测寄生虫载量。此外,我们在刺激全血与可溶性利什曼抗原 [SLA] 前后,测量了单核细胞诱导的 IFN-γ(MIG)和 IFN-γ诱导蛋白 10(IP-10)的生物标志物水平。

主要发现

Kalazar Detect 和 OnSite 分别检测到活动性内脏利什曼病患者的 76%和 64%;分别有 50%和 57%的患者在治疗后仍在这些检测中呈阳性。在研究中居住在流行地区的健康参与者中,只有 1.7%的人同时通过两种 IC 检测呈阳性。另一方面,ELISA 检测的反应性表明,这些人中的 13%存在无症状感染;在 VL 患者中,84%患有活动性疾病的患者在 ELISA 中呈反应性,治疗后,55.5%呈血清阳性。在生活在流行地区的感染个体的血液中,有 37.9%存在利什曼原虫 DNA,而 26.7%的个体中检测到 IP-10 和 MIG 生物标志物。ELISA 和 qPCR 之间的阳性一致性最大。

结论

不同技术的联合应用可以检测无症状感染,但需要进一步研究以开发简单易用的理想生物标志物,用于内脏利什曼病流行地区的临床和现场研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4fc3/6625727/b0c1c976ce40/pntd.0007493.g001.jpg

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