Development of a Novel Multiplex Immunoassay Multi-cruzi for the Serological Confirmation of Chagas Disease
Development of a Novel Multiplex Immunoassay Multi-cruzi for the Serological Confirmation of Chagas Disease
复制标题
DOI：
10.1371/journal.pntd.0004596
复制
发表时间：
2016-04-01
影响因子：
3.8
通讯作者：
Zrein, Maan
中科院分区：
文献类型：
--
作者：
Granjon, Elodie;Dichtel-Danjoy, Marie-Laure;Zrein, Maan
背景查加斯病是由寄生虫克氏锥虫（Trypanosomacruzi）引起的，克氏锥虫是一种原生生物，通过锥虫传播。这种疾病是拉丁美洲的地方病，被卫生组织视为世界上17种被忽视的疾病之一。在欧洲和北美，由于人口向流行地区以外的地区迁移，也发现了输入性病例。T.克氏锥虫感染通常是通过检测T. Cruzi抗原在初始诊断评价或筛选试验（合格或丢弃献血）后，对初始反应性样本进行确认试验。在这项研究中提出的测试的目的是在人类血液样品的感染状态的确认/反驳，并将允许采取适当的临床measures.Methodology/Principal FindingsWe设计了一个新的阵列的12个抗原和打印这些抗原到96孔板。对248份阳性样品进行了T. cruzi，非疫区未筛查献血员94份，阴性献血员49份，假阳性7份，可疑3份。对观察到的反应性进行分析，提出一种决策树算法，该算法可以正确地对所有样品进行分类，并有可能区分假阳性结果和粘性样品。我们观察到，抗体水平（所有抗原的总和）是显着高于PCR阳性的PCR阴性样品在所有研究组与Multi-cruzi.Conclusion/SignificanceThe结果在本研究中描述的表明，Multi-cruzi提高了血清学确认的南美锥虫病。此外，Multi-cruzi检测的“所有抗原之和”可以像PCR信号一样反映患者的寄生虫血症水平，并可以作为纵向随访中寄生虫清除的指标。该检测方法的验证仍然需要在独立的大量充分表征的样品（包括典型的假反应样品，如利什曼病）上进行。
BackgroundChagas disease is due to the parasite Trypanosoma cruzi, a protist disseminated by a Triatome vector. This disease is endemic to Latin America and considered by WHO as one of the 17 world's neglected diseases. In Europe and in North America, imported cases are also detected, due to migration of population outside of the endemic region. Diagnosis of T. cruzi infection is usually made indirectly by the detection of specific antibodies to T. cruzi antigens. Following initial diagnostic evaluation or screening test (qualifying or discarding blood donation), a confirmation test is performed for samples initially reactive. The test presented in this study aims at the confirmation/refutation of the infectious status of human blood samples and will permit taking appropriate clinical measures.Methodology/Principal FindingsWe designed a novel array of twelve antigens and printed these antigens onto 96-well plates. We tested 248 positive samples T. cruzi, 94 unscreened blood donors' samples from non-endemic area, 49 seronegative blood donors, 7 false-positive and 3 doubtful samples. The observed reactivities were analyzed to propose a decision-tree algorithm that correctly classifies all the samples, with the potential to discriminate false-positive results and sticky samples. We observed that antibodies levels (Sum of all antigens) was significantly higher for PCR positive than for PCR negative samples in all studied groups with Multi-cruzi.Conclusion/SignificanceThe results described in this study indicate that the Multi-cruzi improves the serological confirmation of Chagas disease. Moreover the "sum of all antigens" detected by Multi-cruzi could reflect parasitemia level in patients-like PCR signals does-and could serve as an indicator of parasite clearance in longitudinal follow-ups. Validation of this assay is still required on an independent large collection of well characterized samples including typical false-reactive samples such as Leishmaniasis.