Innovations in diagnosis and post-therapeutic monitoring of Chagas disease: Simultaneous flow cytometric detection of IgG1 antibodies anti-live amastigote, anti-live trypomastigote, and anti-fixed epimastigote forms of Trypanosoma cruzi
Innovations in diagnosis and post-therapeutic monitoring of Chagas disease: Simultaneous flow cytometric detection of IgG1 antibodies anti-live amastigote, anti-live trypomastigote, and anti-fixed epimastigote forms of Trypanosoma cruzi
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DOI：
10.1016/j.jim.2014.07.005
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发表时间：
2014-11-01
影响因子：
2.2
通讯作者：
de Lana, Marta
中科院分区：
文献类型：
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作者：
Alessio, Glaucia Diniz;Cortes, Denise Fonseca;de Lana, Marta
这项研究开发了一项显着的方法学创新 (FC-ATE)，能够使用低 (AMA)、中 (TRYPO) 和高 (EPI) 差异荧光染色同时检测克氏锥虫三种进化形式的特异性抗体：活无鞭毛体 (AMA)、活锥鞭毛体 (TRYPO) 和固定表鞭毛体 (EPI)。在区分恰加斯病 (CH) 和非恰加斯病 (NCH) 患者方面表现出色 (100%)。在 FC-ATE 在恰加斯病诊断中的应用中，100% 的 CH 样本的所有三种形式的克氏锥虫的阳性荧光寄生虫 (PPFP) 百分比呈阳性。此外，94% 的 NCH 样本在 AMA 和 TRYPO 中呈现 PPFP 负值，而在 EPI 中则有 88% 的样本呈现负值。 NCH 组中出现假阳性结果的样本属于利什曼病患者。考虑到该技术在恰加斯病治疗后监测中的适用性，100% 的未治疗 (NT) 和治疗未治愈 (TNC) 样本对三种克氏锥虫进化形式呈阳性，而治疗治愈 (TC) 患者样本中 100% 的 AMA 呈阴性，93% 的 TRYPO 呈阴性，96% 的 EPI 呈阴性。 FC-ATE 与使用相同患者 NT、TNC 和 TC 样本的其他两种流式细胞术测试之间的比较表明，尽管观察到方法之间存在分类相关性，但这三种技术呈现出不同的反应性。总而言之，新型 FC-ATE 方法获得的结果在恰加斯病的诊断和治疗后监测中显示出出色的性能。 (C) 2014 Elsevier B.V. 保留所有权利。
This study developed a remarkable methodological innovation (FC-ATE) which enables simultaneous detection of antibodies specific to the three evolutive forms of Trypanosoma cruzi: live amastigote (AMA), live trypomastigote (TRYPO), and fixed epimastigote (EPI) using a differential fluorescence staining as low (AMA), intermediate (TRYPO), and high (EPI). An outstanding performance (100%) was observed in the discrimination of the chagasic (CH) and non-chagasic (NCH) patients. In the applicability of FC-ATE in the diagnosis of Chagas disease, 100% of the CH samples presented positivity in the percentage of positive fluorescent parasites (PPFP) for all the three forms of T. cruzi. Moreover, 94% of the samples of NCH presented negative values of PPFP with AMA and TRYPO, and 88% with EPI. Samples from the NCH group with false-positive results were those belonging to the leishmaniasis patients. Considering the applicability of this technique in post-therapeutic monitoring of Chagas disease, 100% of non-treated (NT) and treated non-cured (TNC) samples were positive with the three T. cruzi evolutive forms, while a percentage of 100% from samples of the treated cured (TC) patients were negative with AMA, 93% with TRYPO and 96% with EPI. The comparison between FC-ATE and two other flow cytometric tests using the same samples of patients NT, TNC and TC showed that the three techniques presented different reactivities, although categorical correlation between the methodologies was observed. Taken together, the results obtained with the novel FC-ATE method have shown an outstanding performance in the diagnosis and post-therapeutic monitoring of Chagas disease. (C) 2014 Elsevier B.V. All rights reserved.