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新疆地区马呼吸道致病病毒的检测及其遗传进化分析 被引量:5

Detection and genetic evolution analysis of equine respiratory disease virus in Xinjiang
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摘要 为了解致汗血马马驹呼吸道疾病的病原,本研究采集新疆乌鲁木齐市南山某汗血马马场82份汗血马的鼻拭子样品(包括4匹患呼吸道疾病的汗血马马驹、6匹健康汗血马马驹和72匹成年健康汗血马),经处理鼻拭子样品、提取病毒RNA、反转录、经PCR扩增及产物纯化后由公司进行高通量测序和宏病毒组学分析。结果显示,与健康马相比,患病马驹鼻拭子样品核酸有1 050条reads注释到马疱疹病毒2型(Equid herpesvirus-2,EHV-2)gB基因,且与EHV-2参考株g B基因序列同源性为91.0%~99.3%,初步表明患病马驹鼻拭子样品携带EHV-2。为验证病毒组学结果,参照EHV-2 G9/92(KM924294)gB基因序列,设计引物,经PCR扩增82份汗血马的鼻拭子样品。PCR结果显示患呼吸道疾病马驹鼻拭子样品均扩增出了EHV-2 gB基因片段,EHV-2阳性率为100%(4/4),而健康马驹和成年汗血马鼻拭子样品均呈阴性结果(0/78),表明EHV-2可能与马驹呼吸道疾病有关。同源性分析结果显示,4株EHV-2之间g B基因和氨基酸序列的同源性分别为94.1%~100%和92.0%~100%,与英国、波兰、瑞士、德国和澳大利亚参考株g B基因和氨基酸序列同源性分别为91.0%~99.3%和89.6%~98.2%。gB氨基酸序列遗传进化分析显示,4株EHV-2与波兰、英国参考株处于同一分支,亲缘关系较近。氨基酸位点分析结果显示,4株EHV-2 g B基因编码氨基酸序列均有一些位点发生了突变,主要位于gB蛋白羧基端,表明4株EHV-2 gB基因具有遗传变异多样性。本研究首次在国内马匹中检测到EHV-2,为EHV-2感染致马驹呼吸道疾病的防控及其流行病学研究提供基础数据。 In order to understand the potential pathogen that causes respiratory diseases of Akhal-teke foals, 82 nasal swabs of Akhal-teke horses(4 Akhal-teke foals suffering from respiratory diseases, 6 healthy Akhal-teke foals, and 72 healthy Akhal-teke horses) were collected from a farm in Nanshan, Urumqi, Xinjiang. Nasal swab samples were used to extract viral RNA and followed by RT-PCR. The purified PCR products were subjected to Illumina sequencing and metaviromics analysis. The resulting metagenomic data of the foals with respiratory disease showed that 1 050 reads were annotated into different genes of Equid herpesvirus(EHV)-2, which shared 91.0%-99.3% nucleotide sequence identity with g B gene of EHV-2 reference strains, indicating that EHV-2 circulated in nasal swabs of sick foals. To confirm the result, EHV-2 G9/92(KM924294) g B gene was used to design PCR primers targeting a 716-nt fragment. PCR result showed EHV-2 positive rate was 100%(4/4) in nasal swab samples of sick Akhal-teke foals, but the virus was not detected in healthy Akhal-teke foals and horses(0/78), indicating that EHV-2 might be associated with foals’ respiratory disease. Homology analysis indicated that g B gene of these four EHV-2 strains shared 94.1%-100% nucleotide sequence identity and 92.0%-100% amino acid sequence identity, and Chinese EHV-2 g B gene shared 91.0%-99.3% nucleotide sequence identity and 89.6%-98.2% amino acid sequence identity with g B gene of EHV-2 reference strains from UK, Switzerland, Germany and Australia. The phylogenetic tree based on the g B protein sequences showed that Chinese EHV-2 were closely related and clustered together with EHV-2 reference strains from UK and Poland. Variations of amino acid residues in g B protein showed that there were some mutations in g B amino acid sequence of four EHV-2 strains, mainly located in the carboxyl end of g B protein, indicated that the four EHV-2 g B genes had genetic diversity. Our data provide first evidence that EHV-2 circulates in horses in China. Our findings provide basic data for the prevention and control of respiratory diseases caused by EHV-2 infection and its epidemiology.
作者 张磊 佟盼盼 任美灵 宋小珍 帕丽旦·努尔兰 贾陈阳 况玲 谢金鑫 ZHANG Lei;TONG Pan-pan;REN Mei-ling;SONG Xiao-zhen;PALIDAN Nuerlan;JIA Chen-yang;KUANG Ling;XIE Jin-xin(College of Veterinary Medicine,Xinjiang Agricultural University,Urumqi 830052,China)
出处 《中国预防兽医学报》 CAS CSCD 北大核心 2021年第7期777-781,共5页 Chinese Journal of Preventive Veterinary Medicine
基金 国家自然科学基金地区基金项目(32060808) 中国博士后科学基金项目(2020T130555、2019M653901XB) 自治区天山创新团队(2020D14005)。
关键词 汗血马马驹 呼吸疾病 病毒组学 马疱疹病毒2型 GB基因 Akhal-teke foal respiratory disease virome equid herpesvirus 2 gB gene
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