Performance of TcI/TcVI/TcII Chagas-Flow ATE-IgG2a for universal and genotype-specific serodiagnosis of Trypanosoma cruzi infection.
Performance of TcI/TcVI/TcII Chagas-Flow ATE-IgG2a for universal and genotype-specific serodiagnosis of Trypanosoma cruzi infection.
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DOI：
10.1371/journal.pntd.0005444
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发表时间：
2017-03
影响因子：
3.8
通讯作者：
de Lana M
中科院分区：
文献类型：
--
作者：
Alessio GD;de Araújo FF;Côrtes DF;Sales Júnior PA;Lima DC;Gomes MS;do Amaral LR;Xavier MA;Teixeira-Carvalho A;Martins-Filho OA;de Lana M
不同的克氏锥虫基因型被认为与恰加斯病的患者管理和治疗反应相关。然而，用于查加斯病的基因型特异性血清学诊断的分型策略仍然不可用，并且需要标准化以用于实际应用。本研究开发了一种新的TcI/TcVI/TcII查加斯流ATE-IgG 2a技术，可用于查加斯原虫的通用和基因型特异性诊断。克氏感染为此，使用无鞭毛体-AMA、锥鞭毛体-TRYPO和外鞭毛体-EPI，在平行批次的TcI、TcVI和TcII靶抗原中，测定了从TcI/Escherichana、TcVI/CL或TcII/Y菌株慢性感染小鼠以及未感染对照小鼠中获得的血清样品的反应性（阳性荧光寄生虫-PPFP的百分比）。结果表明，“α-TcII-TRYPO/1：500，cut-off/PPFP = 20%”是一种较好的T细胞诊断方法。克鲁兹感染（AUC = 1.0，Se和Sp = 100%）。“α-TcI-TRYPO/1：4，000，cut-off/PPFP = 50%"、“α-TcII-AMA/1：1，000，cut-off/PPFP = 40%”和“α-TcVI-EPI/1：1，000，cut-off/PPFP = 45%”的组合属性集显示出分离TcI/Escherichana、TcVI/CL或TcII/Y菌株感染的良好性能。总的来说，TcI/Tcana和TcII/Y株感染的宿主显示与“α-TcI TRYPO”和“α-TcII AMA”相反的反应模式。感染TcVI/CL株的蜱虫呈典型的交织分布。该方法在基因型特异性诊断方面表现良好，当人群/原型场景包括TcI，TcVI和TcII感染时，全局准确率为69%，当仅包括TcI和TcII感染时，全局准确率为94%。本研究还提出了一个受者操作反应性面板，提供了一个可行的工具，从不同的T感染宿主的血清样品进行分类。cruzi基因型，支持这种方法的潜力，通用和基因型特异性诊断T。克氏感染查加斯病仍然是一个重大的公共卫生问题，在全世界感染了600万至700万人。尽管有研究表明不同的临床结果可能与锥虫病的遗传多样性有关，但影响锥虫病临床异质性的因素尚未阐明。克鲁兹分离物。此外，不同T. Cruzi基因型也有报道。查加斯病基因型特异性诊断的分型策略，以确定T。已经开发了包括生物化学和分子方法的Cruzi离散分型单元（DTU），然而这些技术具有局限性。这些方法中的大多数不能直接在生物和临床样品中进行。此外，有人提出，血液中的寄生虫分离株可能不一定代表个体中当前的全套菌株，因为某些菌株可能局限于组织。改进了T.存在于给定宿主中的cruzi DTU可以为临床研究提供有用的工具。在本研究中，我们开发了一种创新的TcI/TcVI/TcII查加斯流ATE-IgG 2a技术，适用于通用和基因型特异性诊断T。cruzi感染，可能有助于增加未来的见解，基因型特异性诊断恰加斯病。
Distinct Trypanosoma cruzi genotypes have been considered relevant for patient management and therapeutic response of Chagas disease. However, typing strategies for genotype-specific serodiagnosis of Chagas disease are still unavailable and requires standardization for practical application. In this study, an innovative TcI/TcVI/TcII Chagas Flow ATE-IgG2a technique was developed with applicability for universal and genotype-specific diagnosis of T. cruzi infection. For this purpose, the reactivity of serum samples (percentage of positive fluorescent parasites-PPFP) obtained from mice chronically infected with TcI/Colombiana, TcVI/CL or TcII/Y strain as well as non-infected controls were determined using amastigote-AMA, trypomastigote-TRYPO and epimastigote-EPI in parallel batches of TcI, TcVI and TcII target antigens. Data demonstrated that “α-TcII-TRYPO/1:500, cut-off/PPFP = 20%” presented an excellent performance for universal diagnosis of T. cruzi infection (AUC = 1.0, Se and Sp = 100%). The combined set of attributes “α-TcI-TRYPO/1:4,000, cut-off/PPFP = 50%”, “α-TcII-AMA/1:1,000, cut-off/PPFP = 40%” and “α-TcVI-EPI/1:1,000, cut-off/PPFP = 45%” showed good performance to segregate infections with TcI/Colombiana, TcVI/CL or TcII/Y strain. Overall, hosts infected with TcI/Colombiana and TcII/Y strains displayed opposite patterns of reactivity with “α-TcI TRYPO” and “α-TcII AMA”. Hosts infected with TcVI/CL strain showed a typical interweaved distribution pattern. The method presented a good performance for genotype-specific diagnosis, with global accuracy of 69% when the population/prototype scenario include TcI, TcVI and TcII infections and 94% when comprise only TcI and TcII infections. This study also proposes a receiver operating reactivity panel, providing a feasible tool to classify serum samples from hosts infected with distinct T. cruzi genotypes, supporting the potential of this method for universal and genotype-specific diagnosis of T. cruzi infection. Chagas disease remains a significant public health issue infecting 6–7 million people worldwide. The factors influencing the clinical heterogeneity of Chagas disease have not been elucidated, although it has been suggested that different clinical outcome may be associated with the genetic diversity of T. cruzi isolates. Moreover, differences in therapeutic response of distinct T. cruzi genotypes have been also reported. Typing strategies for genotype-specific diagnosis of Chagas disease to identify the T. cruzi discrete typing units (DTU) have already been developed, including biochemical and molecular methods, however the techniques have limitations. The majority of these methods can not directly be performed in biological and clinical samples. In addition, it has been proposed that parasite isolates from blood may not necessarily represent the full set of strains current in the individual as some strains can be confined to tissues. The improvement of genotype-specific serology to identify the T. cruzi DTU(s) present in a given host may provide a useful tool for clinical studies. In the present investigation, we developed an innovative TcI/TcVI/TcII Chagas Flow ATE-IgG2a technique with applicability for universal and genotype-specific diagnosis of T. cruzi infection that may contribute to add future insights for genotype-specific diagnosis of Chagas disease.