Epstein J H, Anthony S J
Rev Sci Tech. 2017 Aug;36(2):499-512. doi: 10.20506/rst.36.2.2669.
Emerging diseases are frequently caused by novel or previously unrecognised zoonotic viral pathogens, which tend to originate in and emerge from wildlife. When human or animal cases are first recognised, molecular or serological diagnostic assays specific to them do not yet exist, causing a delay in the identification of an outbreak's aetiologic agent as well as its source. Preparing for the next virus to emerge is a major public health challenge, impeded by a poor understanding of the diversity of potential candidates that exist in wildlife reservoirs. Characterising the diversity of viruses in key wildlife species will help to reduce the time between detection and response in an outbreak situation, and inform public health strategies that reduce the risk of spillover from animal reservoirs. Pathogen discovery techniques such as consensus polymerase chain reaction (cPCR) and next-generation sequencing (NGS) have been used to identify known and novel viruses in animals and humans, but have not been widely used in surveillance programmes. Metagenomic studies have identified novel viruses, new strains of known viruses, and have characterised host microbiomes. While NGS represents an unbiased approach to viral sequence detection, it is constrained by lower sensitivity than conventional PCR, requires substantial bioinformatics capabilities, and is cost prohibitive and therefore not widely available in the regions of the world that are most vulnerable to zoonotic disease emergence. In contrast, consensus PCR uses standard and widely available technologies, has greater sensitivity than NGS, and has also been used to identify novel viruses in wildlife, livestock and humans, though it is limited to detecting target genetic sequences conserved across known groups of viruses. The use of cPCR, in combination, if possible, with NGS and serology, can offer a powerful approach to rapidly identifying aetiologic agents in an outbreak and characterising the virome of key wildlife known to carry zoonotic viruses. Here, the authors review pathogen discovery techniques currently being used in human and animal surveillance programmes and the challenges of using viral discovery to identify novel zoonotic pathogens.
新发疾病通常由新出现的或以前未被识别的人畜共患病毒病原体引起,这些病原体往往起源于野生动物并从野生动物中出现。当首次发现人类或动物病例时,针对这些病例的分子或血清学诊断检测方法尚不存在,这导致在确定疫情的病原体及其来源方面出现延迟。为下一次出现的病毒做好准备是一项重大的公共卫生挑战,而对野生动物宿主中潜在候选病毒多样性的了解不足阻碍了这一挑战的应对。描述关键野生动物物种中病毒的多样性将有助于缩短疫情爆发时从检测到应对的时间,并为降低动物宿主溢出风险的公共卫生策略提供信息。病原体发现技术,如共识聚合酶链反应(cPCR)和下一代测序(NGS),已被用于识别动物和人类中的已知和新型病毒,但尚未在监测计划中广泛使用。宏基因组学研究已经识别出新型病毒、已知病毒的新毒株,并对宿主微生物群进行了特征描述。虽然NGS是一种无偏向性的病毒序列检测方法,但它受到比传统PCR更低的灵敏度的限制,需要大量的生物信息学能力,并且成本高昂,因此在世界上最易出现人畜共患病的地区无法广泛使用。相比之下,共识PCR使用标准且广泛可用的技术,比NGS具有更高的灵敏度,并且也已被用于识别野生动物、家畜和人类中的新型病毒,尽管它仅限于检测已知病毒组中保守的目标基因序列。如果可能的话,将cPCR与NGS和血清学结合使用,可以提供一种强大的方法,用于在疫情爆发时快速识别病原体,并描述已知携带人畜共患病毒的关键野生动物的病毒组特征。在此,作者回顾了目前在人类和动物监测计划中使用的病原体发现技术,以及利用病毒发现来识别新型人畜共患病原体所面临的挑战。