Tripathi Nagesh K, Shrivastava Ambuj, Dash Paban K, Jana Asha M
Defence Research and Development Establishment, Gwalior, India.
Methods Mol Biol. 2011;665:51-64. doi: 10.1007/978-1-60761-817-1_4.
Global incidence of dengue has increased considerably over the past decade. Dengue fever (DF) is a self-limiting disease; however, dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS) are fatal. Since there is no therapy and vaccine against dengue, timely diagnosis is therefore necessary for patient management. Laboratory diagnosis is carried out by virus isolation, demonstration of viral antigen, presence of viral nucleic acid, and antibodies. Further, recombinant dengue envelope protein can be used to detect specific antibodies, both IgG and IgM against all four serotypes of virus using an E. coli vector. The purified protein can then be used for detection of dengue specific IgG or IgM antibodies in patient serum with higher sensitivity and specificity, than that of traditional assays. Molecular detection can be accomplished by a one-step, single-tube, rapid, multiplex, RT-PCR for serotype determination. Despite many advantages of the modern techniques, isolation of virus is still considered as "gold-standard" in dengue diagnosis.
在过去十年中，全球登革热发病率大幅上升。登革热（DF）是一种自限性疾病；然而，登革出血热（DHF）和登革休克综合征（DSS）是致命的。由于目前尚无针对登革热的治疗方法和疫苗，因此及时诊断对于患者管理至关重要。实验室诊断通过病毒分离、病毒抗原检测、病毒核酸检测和抗体检测来进行。此外，重组登革病毒包膜蛋白可用于使用大肠杆菌载体检测针对病毒所有四种血清型的特异性抗体，即IgG和IgM。然后，纯化后的蛋白可用于检测患者血清中登革热特异性IgG或IgM抗体，其灵敏度和特异性均高于传统检测方法。分子检测可通过一步法、单管、快速、多重逆转录聚合酶链反应（RT-PCR）来确定血清型。尽管现代技术有诸多优点，但病毒分离仍被视为登革热诊断的“金标准”。
