摘要
为了探索纳米孔测序技术在动物传染病病原快速诊断中的应用价值,采集1只发生腹泻实验兔的肝脏和脾脏进行核酸提取、建库和纳米孔测序分析,同时对该腹泻的实验兔分别进行细菌分离鉴定和腹泻相关病毒的血清学检测。结果显示:纳米孔测序方法运行3 h后,获得1.9 GB的数据量,检出756521条序列,平均序列长度为2549.9个碱基。根据对测序数据的生物信息学分析表明,引起实验兔腹泻的病原可能为大肠杆菌。对同批送检的6只实验兔采用常规细菌分离鉴定方法在实验兔的心脏血样品、肝脏、肺脏和回盲部内容物中只分离到了大肠杆菌。对腹泻相关病毒的血清学检测结果显示6只送检的兔均为兔轮状病毒阴性。本研究采用纳米孔测序技术对发生腹泻的实验兔进行诊断,其结果与常规实验室检测方法获得的结果相符合,表明纳米孔技术在动物传染病病原快速检测中具有潜在的应用价值。
To explore the application of nanopore sequencing technology in rapid diagnosis of animal diseases,the liver and spleen samples of one experimental rabbit with diarrhea were collected for nucleic acid extraction,library construction and downstream nanopore sequencing analysis.At the same time,isolation and identification of bacteria and serological detection of diarrhea-related viruses were also carried out.The results showed that,using the nanopore sequencing technology,we obtained 1.9 GB of data and detected 756521 sequences with an average length of 2549.9 bases after running for 3 h.According to the bioinformatics analysis of the sequencing data,the pathogen of diarrhea in the experimental rabbit should be Escherichia coli.In addition,Escherichia coli was isolated from the heart blood samples,liver,lung and contents of ileum and cecum of 6 rabbits in the same batch by conventional bacterial isolation and identification.The results of the serological test showed that the 6 rabbits were negative for rotavirus.In this study,the nanopore sequencing technology was used to diagnose diarrhea in experimental rabbits,and the results were consistent with those obtained by conventional laboratory detection,indicating that the nanopore technology had potential application value in the rapid detection of pathogens of animal diseases.
作者
董浩
刘洋
冯育芳
邢进
许中衎
李楠
邢壮壮
王吉
李晓波
陈玲玲
付瑞
梁春南
DONG Hao;LIU Yang;FENG Yufang;XING Jin;XU Zhongkan;LI Nan;XING Zhuangzhuang;WANG Ji;LI Xiaobo;CHEN Lingling;FU Rui;LIANG Chunnan(National Institutes for Food and Drug Control,Beijing 102600,China;China Animal Disease Control Center,Beijing 102600,China)
出处
《畜牧与兽医》
北大核心
2022年第6期126-131,共6页
Animal Husbandry & Veterinary Medicine
基金
国家自然科学基金(31602055)。
关键词
纳米孔测序
细菌分离
大肠杆菌
腹泻
nanopore sequencing
bacterial isolation
E.coli
diarrhea
