摘要
人冠状病毒(Human coronavirus,HCoV)是导致人呼吸道感染的重要病原之一。为了从分子水平上探讨严重急性呼吸道感染(Severe acute respiratory infection,SARI)病例中HCoV-OC43的基因特征,本研究应用实时荧光定量(Real-time)PCR方法对2019年河南省374份SARI病例样本进行HCoV-OC43核酸筛查,并对核酸检测阳性样本进行棘突蛋白(Spike,S)、RNA依赖的RNA聚合酶(RNA-dependent RNA polymerase,RdRp)和核衣壳蛋白(Nucleocapsid,N)三个靶基因扩增和序列测定。结合从GenBank数据库筛选的国内外42条代表序列和本研究所获得的S、RdRp和N三个靶基因全长序列构建基因亲缘性关系树,对本研究所获得的HCoV-OC43样本进行基因型别鉴定和基因特征分析,并对HCoV-OC43重要抗原S蛋白进行氨基酸位点变异分析。结果显示:在374份SARI病例样本中共检测出15份HCoV-OC43核酸阳性样本(15/374,4.01%),并在4份临床样本中获得S、RdRp和N基因全长序列。通过构建基因亲缘性关系树,结果显示有3株属于G基因型,1株属于H基因型。S蛋白氨基酸位点变异分析结果提示,2019年G基因型流行株(包括本研究的3株和美国株USA/MN306041/SC0810/2019)有特异性的3个氨基酸位点变异:L272P、P516S和S902A。2019年H基因型流行株(包括本研究的1株和美国株USA/MN306043/SC0841/2019)特异的氨基酸位点变异为N484D。本研究通过对2019年河南省流行的HCoV-OC43进行基因型别鉴定与基因特征分析,为我国HCoV-OC43的分子变异变迁研究提供了基线数据。
Human coronavirus(HCoV)is one of the important pathogens of human respiratory tract infection. In order to clarify the genetic characteristics of HCoV-OC43 in severe acute respiratory infection(SARI)cases at the molecular level,a total of 374 samples obtained from SARI cases in Henan Province,China,in 2019 were screened for the nucleic acids of HCoV-OC43 by real-time polymerase chain reaction(PCR). Reverse transcription-PCR amplification and sequencing of spike(S),RNA-dependent RNA polymerase(RdRp)and nucleocapsid(N)was carried out in samples with positive detection of the nucleic acids of HCoV-OC43. Upon combination of 42 representative sequences obtained from the GenBank database,phylogenetic trees were constructed based on three full-length sequences of S,RdRp and N genes. The HCoV-OC43 strains obtained from SARI cases were genotyped and the genetic characteristics of three target genes were analyzed. Variations in the amino acids of S protein(an important antigen of HCoV-OC43)were also analyzed. Results showed that15(4.01%)out of 374 samples from SARI cases were positive for HCoV-OC43,and the full-length sequences of S,RdRp and N genes were obtained from 4 out of 15 samples. Based on the phylogenetic trees of these three target genes,three strains belonged to the G genotype and one strain belonged to the H genotype. Analysis of the amino-acid variations of S protein indicated that there were three special sites of amino-acid variation(L272 P,P516 S and S902 A)among the G genotype strains in 2019,including the three strains in our study and USA/MN306041/SC0810/2019. Another special variation in amino acids(N484 D)was found among the H genotype strains in 2019,including one strain in our study and USA/MN306043/SC0841/2019. Based on the genotype identification and genetic characteristics of HCoV-OC43 strains from SARI cases in Henan Province in2019,baseline data for the study of molecular epidemiology of HCoV-OC43 in China have been provided.
作者
孙凯旋
崔爱利
何佩
许文波
张荣波
SUN Kaixuan;CUI Aili;HE Pei;XU Wenbo;ZHANG Rongbo(Medicine College,Anhui University of Science and Technology,Huainan 232001,China;NHC Key Laboratory of Medical Virology and Viral Diseases,National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,WHO WPRO Regional Reference Measles/Rubella Laboratory,Beijing 102206,China)
出处
《病毒学报》
CAS
CSCD
北大核心
2020年第2期236-245,共10页
Chinese Journal of Virology
基金
“十三五”国家科技重大专项课题(项目号:2018ZX10713002),题目:病毒性传染病病原谱和病毒基因变异变迁规律研究
“十三五”国家科技重大专项课题(项目号:2018ZX10713001-003),题目:病毒性自然疫源性传染病症候群病原谱构成及流行规律研究
“十三五”国家科技重大专项“艾滋病和病毒性肝炎等重大传染病防治”(项目号:2018ZX10711001),题目:病毒感染高通量快速检测与应急筛检技术研究。
