2021年银川市4株人呼吸道合胞病毒全基因组基因特征

Gene characteristics of whole genome of four human respiratory syncytial virus in Yinchuan City in 2021

目的 分析2021年银川市人呼吸道合胞病毒(HRSV)全基因组的基因特征。方法 收集2021年银川市2所流感监测哨点医院的流感样标本共1 100份,选择发热并伴有严重呼吸道感染症状的标本共669份,通过实时荧光定量逆转录聚合酶链反应(RT-PCR)法筛选出HRSV阳性标本,对其中符合测序要求的HRSV阳性标本进行全基因组测序。在GenBank数据库下载HRSV全球各基因型别代表株,使用MEGA7软件进行序列比对、亲缘关系进化树构建和遗传距离的计算分析,使用DNAStar软件中MegAlign分析HRSV测序株序列核苷酸(氨基酸)的同源性和氨基酸变异。结果 669份全年龄段的鼻咽拭子标本,HRSV阳性标本60份,阳性率为8.97%。其中有4份HRSV阳性标本全基因组测序成功,亲缘关系进化树显示,2株HRSVA为ON1型,2株HRSVB为BA9型。2条HRSVA全长序列均与MN306029.1/USA/2019/ON1全长序列的核苷酸(氨基酸)同源性最高,2条HRSVB G蛋白基因序列均与MW160815.1/Australia/2017/BA9的G蛋白基因序列核苷酸(氨基酸)同源性最高。氨基酸变异分析结果表明,2条HRSVA在第113、131、178、257、258、266位均发生了突变,分别为T113I、V131D、N178G、E257K、H258Q、H266L,YinChuanRSVA-01在第134位发生K突变为R(K134R),2条HRSVB在第159位和第160位均出现氨基酸缺失,在G蛋白第二高变区的第254、270、276、290位均发生T突变为I(T254I、T270I、T276I、T290I)。结论 2021年银川市HRSV出现了RSVA基因型和RSVB基因型的共同流行,以及多个氨基酸位点的变异。

Objective To analyze the gene characteristics of the whole genome of human respiratory syncytial virus(HRSV) in Yinchuan City in 2021.Methods A total of 1 100 influenza-like specimens from two influenza surveillance sentinel hospitals in Yinchuan City in 2021 were collected,669 specimens from patients with fever and severe respiratory tract infection symptoms were selected,HRSV positive specimens were screened by real-time fluorescence quantitative reverse transcription polymerase chain reaction(RT-PCR),HRSV positive specimens that met the sequencing requirements were performed whole genome sequencing.HRSV representative strains of various genotypes in the world were downloaded from GenBanks database,sequence alignment,phylogenetic tree construction as well as genetic distance calculation and analysis were conducted by MEGA7 software,homology of nucleotide(amino acid) and amino acid variation of HRSV sequenced strains were analyzed by MegAalign in DNAStar software.Results Among 669 nasopharyngeal swabs from patients of all ages,60 were HRSV positive,with a positive rate of 8.97%,whole genome of 4 HRSV positive specimens were successfully sequenced,phylogenetic tree showed that 2 HRSVA strains were type ON1 and 2 HRSVB strains were type BA9.Two HRSVA full-length sequences had the highest nucleotide(amino acid) homology with MN306029.1/USA/2019/ON1 full-length sequence,two HRSVB G protein gene sequences had the highest nucleotide(amino acid) homology with MW160815.1/Australia/2017/BA9 G protein gene sequence.Amino acid variation analysis showed that 2 HRSVA were mutated at 113,131,178,257,258 and 266 loci,respectively T113 I,V131 D,N178 G,E257 K,H258 Q,and H266 L,YinChuanRSVA-01 had K mutation to R(K134 R) at position 134,two HRSVB showed amino acid deletion at 159 and 160 loci,and T mutation to I(T254 I,T270 I,T276 I,T290 I) at 254,270,276 and 290 loci of the second hypervariable region of G protein.Conclusion In Yinchuan City in 2021,there was a common prevalence of RSVA genotype and RSVB genotype of HRSV as well as variations in multiple amino acid loci.