新发病毒检测方法与测序技术的研究进展及应用

doi:10.11843/j.issn.0366-6964.2024.12.007

摘要/Abstract

摘要：

近年来，随着人口的快速增长，国际贸易日益频繁，由未知病毒感染引起的新发传染病对人类健康、畜牧业的发展以及公共卫生安全的威胁正在不断增加。因此快速、精确地监测和检测新发病毒是控制和预防传染病疫情最有效的方法之一。传统的病毒检测方法操作简单、技术要求低，但在准确性、时效性等方面存在较多局限。而利用适宜的分子生物学方法及测序技术可直接从临床样本中检测未知病毒，这对于病原的快速识别并及时发现新型病毒具有重要意义。本文综述了近年来应用于新发病毒检测的方法及测序技术的基本原理、发展情况和应用领域，分析比较其差异及优缺点，以供读者参考。

关键词: 新发病毒, 检测方法, 测序技术, 应用

Abstract:

In recent years, given the rapid increase in population and the frequency of international trade, the risks associated with emerging infectious diseases caused by unknown viral agents have heightened, posing threats to human health, animal husbandry, and public health. Therefore, the ability to rapidly and accurately monitor and detect emerging viruses is among the most effective strategies for controlling and preventing the outbreak of infectious diseases. While traditional methods for detecting virus are simple to operate and require minimal technical expertise, they are limited in terms of accuracy and timeliness. Unknown viruses can be directly detected from clinical samples by using appropriate molecular biological methods and sequencing technology, which is of great significance for the rapid identification of pathogens and the timely discovery of new viruses. This article reviews the basic principles, evolution and application fields of the methods and sequencing technologies used in the detection of emerging viruses in recent years. It also analyzes and compares their differences, advantages and disadvantages, offering valuable reference for the readers.

Key words: emerging virus, detection methods, sequencing technology, applications

中图分类号:

S854.4

吕岱玥, 陈延飞, 翟天舒, 曹胜波, 薛青红. 新发病毒检测方法与测序技术的研究进展及应用[J]. 畜牧兽医学报, 2024, 55(12): 5398-5411.

LÜ Daiyue, CHEN Yanfei, ZHAI Tianshu, CAO Shengbo, XUE Qinghong. Research Progress and Application of Emerging Virus Detection Methods and Sequencing Technology[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(12): 5398-5411.

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方法 Method	主要原理 Main principles	优、缺点 Advantage, disadvantage	主要应用 Main application
变性寡核苷酸引物PCR DOP-PCR	部分随机引物法	对模板质量要求低，操作简单。但扩增产物平均长度较短，基因组覆盖率低	可鉴定未知DNA、RNA 病毒序列
滚环复制技术 RCA	自然滚环复制的简化衍生	高灵敏度，高序列特异性，无需制定引物，高通量，操作方便。但合成费用较高，信号检测时存在背景干扰问题	可鉴定未知环状DNA 病毒序列
多重置换扩增技术 MDA	φ29 DNA聚合酶的链置换机制	技术操作简单、产物质量稳定。但对模板质量要求高	可鉴定未知线状DNA 病毒序列
代表性差异分析 RDA	基因组的消减杂交	具有较高的灵敏度，无需文库的筛选，可重复性好。但操作过程相对繁琐复杂，实验周期长，会出现假阳性结果	可鉴定未知DNA、RNA 病毒序列
序列非依赖的单引物扩增 SISPA	平末端连接扩增	不依赖于组织培养，可用于不同类型的临床样本。但样品中存在其他污染序列，而且价格昂贵	可鉴定未知DNA、RNA 病毒序列
cDNA扩增片段长度多态性 VIDISCA	一种SISPA衍生方法	灵敏度较高	可鉴定未知DNA、RNA 病毒序列
随机引物PCR AP-PCR	随机引物法	对模板DNA的纯度要求不高；技术简单；灵敏度高，可提供丰富的多态性但随机引物PCR技术易受外界因素的影响，且扩增产物的稳定性较低	可鉴定未知DNA、RNA 病毒序列
表达cDNA文库 cDNA library	特异性表位筛选与鉴定	耗时长且阳性率低	可鉴定未知RNA 病毒序列

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