Accomplishing the genotype-specific serodiagnosis of single and dual Trypanosoma cruzi infections by flow cytometry Chagas-Flow ATE-IgG2a.
Accomplishing the genotype-specific serodiagnosis of single and dual Trypanosoma cruzi infections by flow cytometry Chagas-Flow ATE-IgG2a.
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DOI：
10.1371/journal.pntd.0006140
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发表时间：
2018-03
影响因子：
3.8
通讯作者：
Martins-Filho OA
中科院分区：
文献类型：
--
作者：
Alessio GD;de Araújo FF;Sales Júnior PA;Gomes MS;Amaral LRD;Pascoal Xavier MA;Teixeira-Carvalho A;de Lana M;Martins-Filho OA
目前可用于克氏锥虫感染基因型特异性诊断的方法仍然存在相关局限性，特别是在识别混合感染方面。在本研究中，我们评估了 Chagas-Flow ATE-IgG2a 测试对单基因型和双基因型特异性克氏锥虫感染的早期和晚期鉴别诊断的性能。对克氏锥虫感染早期和晚期瑞士小鼠的血清样本进行平行批次分析，以进行单一（TcI、TcVI 或 TcII）和双重（TcI+TcVI、TcVI+TcII 或 TcII+TcI）感染的基因型特异性诊断。在特定反向血清稀释度（500 至 64,000）下，对 TcI、TcVI 和 TcII 靶抗原（包括无鞭毛体 (AI/AVI/AII)、锥鞭毛体 (TI/TVI/TII) 和表鞭毛体 (EI/EVI/EII)）的内在反应性，用于为“早期”与“晚期”、“单与“双”和结果表明，选择性属性集“EII 500/EI 2,000/AII 500”能够提供高质量的准确度（81%）来分离克氏锥虫感染的早期和晚期阶段。“TI 2,000/AI 1,000/EII 1,000”和“TI 8,000/AII 32,000”。分别提出了区分早期（85%）和晚期（84%）克氏锥虫单一感染和双重感染的表达分数。此外，属性“TI 4,000/TVI 500/TII 1,000”、“TI 16,000/EI 2,000/EII 2,000/AI 500/TVI 500”在早期阶段的基因型特异性诊断中表现出良好的性能。此外，属性“TI 4,000/AII 1,000/EVI 1,000”、“TI 64,000/AVI 500/AI 2,000/AII 1,000/EII 4,000”在单次感染（69%）的晚期阶段表现出中等的基因型特异性诊断性能。这些结果为恰加斯流 ATE-IgG2a 方法在人类基因型特异性诊断中的应用提供了新的视角，为流行病学调查以及克氏锥虫病的临床和治疗后监测做出了相关贡献。显示出巨大的遗传多样性，并被细分为六个 DTU（离散分型单位），称为 TcI-TcVI，这种遗传和生物学变异，加上宿主的自然再感染，可能在该疾病的临床和流行病学特征中发挥重要作用。此外，感染不同克氏锥虫基因型的宿主表现出不同的治疗反应，因此，开发针对克氏锥虫感染的基因型特异性诊断的新方法对于临床和流行病学非常重要。生化和分子方法用于此目的，但是，这些技术具有方法学局限性，此外，用于恰加斯病基因型特异性诊断的标准化血清学方法存在抗原局限性，并且也不能评估混合感染的血清样本的反应性。根据我们之前的结果，在本次研究中，我们评估了 Chagas-Flow ATE-IgG2a 在单次和双次克氏锥虫感染早期和晚期基因型特异性诊断中的适用性。
The methods currently available for genotype-specific diagnosis of T. cruzi infection still present relevant limitations, especially to identify mixed infection. In the present investigation, we have evaluated the performance of Chagas-Flow ATE-IgG2a test for early and late differential diagnosis of single and dual genotype-specific T. cruzi infections. Serum samples from Swiss mice at early and late stages of T. cruzi infection were assayed in parallel batches for genotype-specific diagnosis of single (TcI, TcVI or TcII) and dual (TcI+TcVI, TcVI+TcII or TcII+TcI) infections. The intrinsic reactivity to TcI, TcVI and TcII target antigens, including amastigote (AI/AVI/AII), trypomastigote-(TI/TVI/TII) and epimastigote (EI/EVI/EII), at specific reverse of serum dilutions (500 to 64,000), was employed to provide reliable decision-trees for “early” vs “late”, “single vs “dual” and “genotype-specific” serology. The results demonstrated that selective set of attributes “EII 500/EI 2,000/AII 500” were able to provide high-quality accuracy (81%) to segregate early and late stages of T. cruzi infection. The sets “TI 2,000/AI 1,000/EII 1,000” and “TI 8,000/AII 32,000” presented expressive scores to discriminate single from dual T. cruzi infections at early (85%) and late stages (84%), respectively. Moreover, the attributes “TI 4,000/TVI 500/TII 1,000”, “TI 16,000/EI 2,000/EII 2,000/AI 500/TVI 500” showed good performance for genotype-specific diagnosis at early stage of single (72%) and dual (80%) T. cruzi infections, respectively. In addition, the attributes “TI 4,000/AII 1,000/EVI 1,000”, “TI 64,000/AVI 500/AI 2,000/AII 1,000/EII 4,000” showed moderate performance for genotype-specific diagnosis at late stage of single (69%) and dual (76%) T. cruzi infections, respectively. The sets of decision-trees were assembled to construct a sequential algorithm with expressive accuracy (81%) for serological diagnosis of T. cruzi infection. These findings engender new perspectives for the application of Chagas-Flow ATE-IgG2a method for genotype-specific diagnosis in humans, with relevant contributions for epidemiological surveys as well as clinical and post-therapeutic monitoring of Chagas disease. Trypanosoma cruzi shows great genetic diversity, and was subdivided into six DTUs (Discrete Typing Units), named TcI-TcVI. This genetic and biological variability, coupled with natural reinfection of hosts, may play an important role in the clinical and epidemiological features of the disease. Furthermore, hosts infected with different T. cruzi genotypes demonstrated distinct therapeutic response. Thus, the development of new methods for genotype-specific diagnosis of T. cruzi infection is very important for clinical and epidemiological studies and for post-therapeutic monitoring of patients treated. Biochemical and molecular methods are used for the purpose, however, these techniques have methodological limitations. In addition, the standardized serological methods for genotype-specific diagnosis of Chagas disease present antigenic limitations and also do not evaluate the reactivity of serum samples from mixed infections. In order to overcome these challenges, our group developed the Chagas-Flow ATE-IgG2a technique with good performance for universal and genotype-specific diagnosis of single T. cruzi infection in the chronic phase. Based on our previous results, in the present investigation, we evaluated the applicability of Chagas-Flow ATE-IgG2a in the genotype-specific diagnosis at early and late stages for single and dual T. cruzi infections.
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影响因子：
3.8
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
通讯作者：
de Lana M
DOI：
10.1590/s0074-02762011000800009
发表时间：
2011-12-01
期刊：
Memórias do Instituto Oswaldo Cruz
影响因子：
--
作者：
Andrade, Sonia Gumes;Campos, Rozália Figueira;Macedo, Andréa Mara
通讯作者：
Macedo, Andréa Mara
影响因子：
2.7
作者：
Araujo, Catarina A. C.;Cabello, Pedro H.;Jansen, Ana M.
通讯作者：
Jansen, Ana M.
影响因子：
2.2
作者：
Alessio, Glaucia Diniz;Cortes, Denise Fonseca;de Lana, Marta
通讯作者：
de Lana, Marta
影响因子：
2.1
作者：
Bosseno, MF;Telleria, J;Breniere, SF
通讯作者：
Breniere, SF