Gaibani Paolo, Landini Maria Poala, Sambri Vittorio
Regional Reference Centre for Microbiological Emergencies (CRREM), Unit of Microbiology, S. Orsola-Malpighi Hospital, Via G. Massarenti 9-40138, Bologna, Italy.
DIMES, University of Bologna, Bologna, Italy.
Methods Mol Biol. 2016;1426:63-73. doi: 10.1007/978-1-4939-3618-2_6.
This chapter presents the most commonly used serological methods for the diagnosis of Chikungunya virus (CHIKV) infection in humans. CHIKV is a mosquito-borne Alphavirus widely distributed in the tropical and subtropical regions of Africa, Asia, and America. CHIKV infection in human causes acute febrile illness frequently accompanied by severe joint pain. Most of the infected patients may develop chronic arthralgia that may persist for several months or years. Laboratory diagnosis of CHIKV infection is mainly based on molecular and serological tests. The serological tests represent a valuable tool for diagnosis and epidemiological studies. Enzyme-linked immunosorbent assay (ELISA) and immunofluorescence assay (IFA) are simple, rapid, and sensitive techniques widely used for the diagnosis of CHIKV infection. However, these methods represent a screening tool and often require confirmation by a second-line assays. Serum virus neutralization assay is more specific than ELISA and IFA tests and is considered a confirmatory test. Neutralization assay is employed to determine the titer of virus neutralizing antibodies against CHIKV in patients' sera. The basis of microneutralization assay (MNA), results interpretation, and procedures will be illustrated in this chapter.
本章介绍了用于诊断人类基孔肯雅病毒(CHIKV)感染的最常用血清学方法。CHIKV是一种由蚊子传播的甲病毒,广泛分布于非洲、亚洲和美洲的热带和亚热带地区。人类感染CHIKV会引发急性发热疾病,并常伴有严重的关节疼痛。大多数感染患者可能会出现慢性关节痛,这种疼痛可能会持续数月或数年。CHIKV感染的实验室诊断主要基于分子和血清学检测。血清学检测是诊断和流行病学研究的重要工具。酶联免疫吸附测定(ELISA)和免疫荧光测定(IFA)是简单、快速且灵敏的技术,广泛用于CHIKV感染的诊断。然而,这些方法只是筛查工具,通常需要通过二线检测进行确认。血清病毒中和试验比ELISA和IFA检测更具特异性,被视为确诊试验。中和试验用于测定患者血清中针对CHIKV的病毒中和抗体滴度。本章将阐述微量中和试验(MNA)的原理、结果解读及操作步骤。