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登革热的血清学诊断

Serological Diagnosis of Dengue.

作者信息

Lima Monique R Q, Nunes Priscila C G, Dos Santos Flávia B

机构信息

Laboratório Estratégico de Diagnóstico (LED), Centro de Desenvolvimento Científico, Instituto Butantan, São Paulo, Brazil.

Laboratório Municipal de Saúde Pública (LASP), Laboratório de Virologia e Biotério, Subsecretaria de Vigilância, Fiscalização Sanitária e Controle de Zoonoses, Rio de Janeiro, Brazil.

出版信息

Methods Mol Biol. 2022;2409:173-196. doi: 10.1007/978-1-0716-1879-0_12.

Abstract

A reliable and specific diagnosis is imperative in viral diagnosis, both for clinical management and surveillance, and to ensure that early treatment and control measures are carried out. The number of days of illness is important to choose the most appropriate method to be used and for the correct interpretation of the results obtained. Specific IgM is elicited after that period, indicating an active infection and usually lasts up to 3 months. However, in DENV secondary infections, IgM levels may be significantly lower or undetectable. After 10-12 days, a lifetime specific IgG is produced. Routinely, the laboratory diagnosis of DENV infections can be performed by viral isolation and/or detection of viral nucleic acid, serological assays for the detection of specific antibodies (IgM/IgG), antigen (NS1) and the detection of viral antigens in tissues, which are suitable during certain phases of the disease. For serological diagnosis, serum, plasma, or cerebrospinal fluid (CSF) samples may be investigated. If the test is carried out a few days after collection, the specimens can be stored at 4 °C, since the immunoglobulins are stable in serum or plasma. If the storage period is extended, the material must be kept at -20 °C or -70 °C. In serology, several methods can be used to detect specific viral antigens and/or antibodies, produced by the host in response to DENV infection. Routinely, serological tests include the hemagglutination inhibition (HI) assay, the plaque reduction neutralizing test (PRNT), the gold standard assay for dengue immune response characterization, and ELISAs to detect IgM (MAC-ELISA) and IgG (IgG-ELISA).

摘要

对于病毒诊断而言,可靠且特异的诊断至关重要,这对于临床管理和监测,以及确保实施早期治疗和控制措施都很关键。发病天数对于选择最合适的检测方法以及正确解读检测结果很重要。在此期间之后会产生特异性IgM,表明存在活动性感染,且通常可持续长达3个月。然而,在登革病毒二次感染中,IgM水平可能显著降低或无法检测到。10 - 12天后会产生终身性的特异性IgG。常规情况下,登革病毒感染的实验室诊断可通过病毒分离和/或检测病毒核酸、用于检测特异性抗体(IgM/IgG)和抗原(NS1)的血清学检测以及在组织中检测病毒抗原进行,这些方法在疾病的特定阶段适用。对于血清学诊断,可对血清、血浆或脑脊液(CSF)样本进行检测。如果在采集后几天内进行检测,样本可储存在4℃,因为免疫球蛋白在血清或血浆中稳定。如果储存期延长,样本必须保存在-20℃或-70℃。在血清学中,可使用多种方法来检测宿主针对登革病毒感染产生的特异性病毒抗原和/或抗体。常规的血清学检测包括血凝抑制(HI)试验、蚀斑减少中和试验(PRNT),这是用于登革免疫反应特征鉴定的金标准试验,以及用于检测IgM(MAC - ELISA)和IgG(IgG - ELISA)的酶联免疫吸附测定。

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