Next-generation ELISA diagnostic assay for Chagas Disease based on the combination of short peptidic epitopes.
Next-generation ELISA diagnostic assay for Chagas Disease based on the combination of short peptidic epitopes.
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DOI：
10.1371/journal.pntd.0005972
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发表时间：
2017-10
影响因子：
3.8
通讯作者：
Agüero F
中科院分区：
文献类型：
--
作者：
Mucci J;Carmona SJ;Volcovich R;Altcheh J;Bracamonte E;Marco JD;Nielsen M;Buscaglia CA;Agüero F
由原生动物克氏锥虫引起的恰加斯病是拉丁美洲的一个重大卫生和经济问题，目前还没有疫苗或适当药物可用于大规模公共卫生干预。准确诊断对于早期识别和跟踪病媒传播病例以及防止通过输血和器官移植传播疾病至关重要。常规诊断采用血清学方法，其中一些方法需要生产寄生虫裂解物、寄生虫抗原部分或纯化的重组抗原。虽然现有的血清学试验结果令人满意，但生产可靠的试剂仍然费力且昂贵。跨越线性b细胞表位的短肽已被证明是广泛疾病的理想血清诊断试剂。最近，我们利用高密度肽芯片对克氏锥虫线性b细胞表位进行了大规模筛选，鉴定出数百个与慢性恰加斯病相关的新序列特征。在这里，我们对27个选定的表位进行了血清学评估，并对它们在一种新的多肽诊断方法中的应用进行了评估。最后对199份血清样本（恰加斯病阳性和阴性，包括利什曼病阳性受试者的血清）以ELISA格式对其中7种多肽的组合进行了评估。多肽制剂具有较高的诊断效能，敏感性为96.3%，特异性为99.15%。因此，使用合成肽作为诊断工具在南美锥虫病诊断中是一种有吸引力的替代方法。由克氏锥虫寄生虫引起的恰加斯病是一种终生致残的疾病，在整个拉丁美洲具有重大意义，也是对全球公共卫生的一个新威胁。诊断测试是支持疾病监测并最终帮助阻止寄生虫传播的关键工具。然而，目前可用的诊断方法有一些局限性。鉴定具有改进诊断特征的新型生物标志物是一个主要的优先事项。最近，我们进行了大规模的筛选，寻找新的克氏锥虫抗原使用短肽显示在高密度固体载体上。这导致了数百个新的抗原表位的鉴定。在这项工作中，我们验证了其中27种针对人类血清样本扩展面板的血清诊断性能。基于这一分析，我们开发了一种原理验证的多重诊断试剂盒，通过组合不同的验证活性肽。总的来说，我们的数据支持高密度肽微阵列在快速识别和定位表位方面的适用性，这些表位可以很容易地转化为诊断恰加斯病的新颖有用的工具。
Chagas Disease, caused by the protozoan Trypanosoma cruzi, is a major health and economic problem in Latin America for which no vaccine or appropriate drugs for large-scale public health interventions are yet available. Accurate diagnosis is essential for the early identification and follow up of vector-borne cases and to prevent transmission of the disease by way of blood transfusions and organ transplantation. Diagnosis is routinely performed using serological methods, some of which require the production of parasite lysates, parasite antigenic fractions or purified recombinant antigens. Although available serological tests give satisfactory results, the production of reliable reagents remains laborious and expensive. Short peptides spanning linear B-cell epitopes have proven ideal serodiagnostic reagents in a wide range of diseases. Recently, we have conducted a large-scale screening of T. cruzi linear B-cell epitopes using high-density peptide chips, leading to the identification of several hundred novel sequence signatures associated to chronic Chagas Disease. Here, we performed a serological assessment of 27 selected epitopes and of their use in a novel multipeptide-based diagnostic method. A combination of 7 of these peptides were finally evaluated in ELISA format against a panel of 199 sera samples (Chagas-positive and negative, including sera from Leishmaniasis-positive subjects). The multipeptide formulation displayed a high diagnostic performance, with a sensitivity of 96.3% and a specificity of 99.15%. Therefore, the use of synthetic peptides as diagnostic tools are an attractive alternative in Chagas’ disease diagnosis. Chagas disease, caused by the parasite Trypanosoma cruzi, is a life-long and debilitating illness of major significance throughout Latin America, and an emergent threat to global public health. Diagnostic tests are key tools to support disease surveillance, and to ultimately help stop transmission of the parasite. However currently available diagnostic methods have several limitations. Identification of novel biomarkers with improved diagnostic characteristics is a main priority. Recently, we conducted a large-scale screening looking for new T. cruzi antigens using short peptides displayed on a solid support at high-density. This led to the identification of several hundred novel antigenic epitopes. In this work we validated the serodiagnostic performance of 27 of these against an extended panel of human serum samples. Based on this analysis, we developed a proof-of-principle multiplex diagnostic kit by combining different validated reactive peptides. Overall, our data support the applicability of high-density peptide microarrays for the rapid identification and mapping epitopes that could be readily translated into novel and useful tools for diagnosis of Chagas disease.
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影响因子：
--
作者：
Dai, Jianfang;Jiang, Min;Si, Jin
通讯作者：
Si, Jin
影响因子：
3.8
作者：
dos Santos SL;Freitas LM;Lobo FP;Rodrigues-Luiz GF;Mendes TA;Oliveira AC;Andrade LO;Chiari E;Gazzinelli RT;Teixeira SM;Fujiwara RT;Bartholomeu DC
通讯作者：
Bartholomeu DC
DOI：
10.1073/pnas.89.4.1239
发表时间：
1992-02-15
影响因子：
11.1
作者：
BURNS, JM;SHREFFLER, WG;REED, SG
通讯作者：
REED, SG
影响因子：
1.5
作者：
Campo, V;Di Noia, JM;Frasch, ACC
通讯作者：
Frasch, ACC
影响因子：
14.9
作者：
Bartholomeu, Daniella C.;Cerqueira, Gustavo C.;El-Sayed, Najib M.
通讯作者：
El-Sayed, Najib M.