摘要
本研究旨在借助宏基因组测序分析技术快速发现引起濒危亚洲象死亡的可能病原,再结合常规诊断技术,对大象病因进行分析和确诊,为大象相关疫病的快速诊断和预防提供参考.对死亡大象进行尸检,取心、肝、脾、肺、肾、淋巴结、肌肉、胃、十二指肠、空肠、结肠、盲肠、胰腺等器官制备切片,进行HE染色和病理分析;取腹股沟淋巴结进行宏基因组测序分析;使用特异性PCR方法对特征基因进行扩增和测序,经序列比对绘制进化树,分析其与其他毒株的进化关系.结果显示,死亡大象的剖检和病例切片结果符合象亲内皮疱疹病毒感染的特征,宏基因组分析结果显示,大象的腹股沟淋巴结中含有大量的象β疱疹病毒1型(即亲内皮疱疹病毒).该大象死亡的原因可能是象亲内皮疱疹病毒感染所致,宏基因组技术对大象亲内皮疱疹病毒感染以及其他濒危动物病例的快速诊断具有良好的应用前景.
Abstract
This study aims to use metagenomic sequencing analysis technology to quickly diagnose the possible pathogens causing the death of an Asian elephant,and then combine with conven-tional diagnostic techniques to analyze and confirm the etiological cause of elephant diseases,so as to provide reference for rapid diagnosis and prevention of elephant-related diseases.Autopsies were performed on dead elephant,and organs such as the heart,liver,spleen,lungs,kidneys,lymph nodes,muscles,stomach,duodenum,jejunum,colon,cecum,and pancreas were collect-ed and sectioned for HE staining and pathological analysis;Metagenomic sequencing analysis was performed on the inguinal lymph nodes;species-specific PCR were used to amplify and sequence the characteristic genes,and an evolutionary tree was drawn to analyze its phylogenetic relation-ship with other strains.Data showed that necropsy and case biopsy results of dead elephant are consistent with the clinical characteristics of endothelial herpesvirus infection in elephants.Met-agenomic analysis detected a large number of elephant betaherpesvirus type 1(i.e.,parent endo-thelial herpes virus)in the inguinal lymph nodes of elephants.The death of the elephant was probably caused by endothelial herpesvirus infection.Metagenomic technology offers valuable application prospects for the rapid diagnosis of endothelial herpesvirus infection in elephants as well as other endangered animal clinical infectious cases.
基金项目
重大外来动物疫病阻断与防控技术研发项目(2022YFD1800500)
维普
万方数据