Dengue is an acute febrile illness caused by positive-sense single-stranded RNA virus, belonging to the family Flaviviridae and genus Flavivirus. Transmission of virus among the individuals occurred by blood-feeding Aedes mosquitoes. This virus has four serotypes differentiated on the basis of antibody neutralization assay. At present, there is no particular treatment or vaccine candidate available for dengue infection. Approximately 3.9 billion human populations are at risk of dengue virus (DENV) infection. Thus, precise diagnosis of dengue at the early stage is very essential for disease control and effective therapy in order to treat or prevent severe complications. Indeed, the accurate diagnosis of DENV remains a problem because of low detection accuracy along with high testing price. Sensitivity and specificity of available kits vary from test to test, and cross-reactivity with other Flavivirus is a challenging issue for diagnosis. In this study, linear epitopes of envelope (E) and NS1 proteins were identified to diagnose the DENV. Whole protein sequences of E and NS1 of DENV were obtained from UniProtKB database. On the basis of algorithm prediction from DNASTAR, BCEPRED, and IEDB data resources, twelve peptides of E (EP1 to EP12) and eight peptides of NS1 (NS1-1 to NS1-8) were selected, which were common in all serotypes. Sequence homologies of peptides with other Flavivirus were checked by Multiple Sequence Alignment Tool ClustalX2. Peptide sequences were synthesized chemically by solid-phase peptide synthesis technique. Dengue-specific IgM and IgG (secondary response) antibodies in the patient’s antisera were tested with the peptides using ELISA protocol. Peptides EP1, EP2, EP4, EP7, EP10, and EP12 of E protein and NS1-1, NS1-3, NS1-4, NS1-7, and NS1-8 of NS1 protein were considered the best immunoreactive peptides with the sensitivity (73.33-96.66%) and specificity (82.14-100%). Such peptides together can be used to construct the multiple antigen peptides (MAP) or multiplexed microbeads for designing a precise, cost-effective, and easy-to-make peptide-based immunodiagnostic kit for DENV detection.

中文翻译：

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通过信封和NS1蛋白的肽序列检测登革热病毒特异性IgM和IgG抗体，以进行血清学鉴定。

登革热是由正链单链RNA病毒引起的一种急性发热性疾病，属于黄病毒科和黄病毒科。摄食伊蚊导致病毒在个体之间传播蚊子。根据抗体中和分析，该病毒具有四种血清型。目前，尚无可用于登革热感染的特殊治疗方法或候选疫苗。大约39亿人口面临登革热病毒（DENV）感染的危险。因此，对登革热的早期精确诊断对于控制或预防严重并发症的疾病控制和有效治疗至关重要。实际上，由于低的检测精度和高昂的测试价格，DENV的准确诊断仍然是一个问题。可用试剂盒的灵敏度和特异性因测试而异，并且与其他黄病毒有交叉反应对于诊断是一个具有挑战性的问题。在这项研究中，鉴定了包膜（E）和NS1蛋白的线性表位来诊断DENV。从UniProtKB数据库获得DENV的E和NS1的全蛋白序列。在根据DNASTAR，BCEPRED和IEDB数据资源进行算法预测的基础上，选择了12种E肽（EP1至EP12）和8种NS1肽（NS1-1至NS1-8），这在所有血清型中都是常见的。多肽与其他黄病毒的序列同源性通过多序列比对工具ClustalX2进行了检查。通过固相肽合成技术化学合成肽序列。使用ELISA方案，使用肽段测试患者抗血清中的登革热特异性I​​gM和IgG（第二反应）抗体。E蛋白的肽EP1，EP2，EP4，EP7，EP10和EP12和NS1蛋白的NS1-1，NS1-3，NS1-4，NS1-7和NS1-8被认为是最佳的免疫反应性肽，具有以下敏感性： 73.33-96.66％）和特异性（82.14-100％）。此类肽可一起用于构建多种抗原肽（MAP）或多重微珠，以设计用于DENV检测的精确，经济高效且易于制造的基于肽的免疫诊断试剂盒。