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对存于中国疾病控制预防控制中心病毒病预防控制所病毒性腹泻室的一例卢龙G3型仔猪GARV LLZ212的主要基因片段VP7、VP4、VP6、VP2和NSP2-5进行扩增,确定其基因型,并探讨其与当地人G3型GARV的进化关系。采用逆转录-聚合酶链式反应(Reverse transcription-polymerase chain reaction,RT-PCR),对LLZ212标本及6例当地儿童G3型GARV阳性标本的VP7、VP4、VP6、VP2和NSP2-5进行扩增,并进行序列测定。应用A组轮状病毒自动分型工具(RotaC v2.0)对各个片段进行分型。应用DNAStar和MEGA5.0软件包与GenBank上的参考株进行同源性和系统进化分析。卢龙猪LLZ212基因型为G3-P[8]-I5-C1-N1-T1-E1-H1,6株人GARV基因型均为G3-P[8]-I1-C1-N1-T1-E1-H1。LLZ212基因片段VP7、VP4、NSP4和NSP5分别与人GARV E885、CMH014/07、Wa和RMC321株同源性最高,且进化树在同一亚枝内;LLZ212基因片段VP6、NSP2和NSP3均与猪GARV PRG921株同源性最高,基因片段VP2与猪GARV OSU株同源性最高,且进化树在同一亚枝内。确定一株罕见G3-P[8]-I5-C1-N1-T1-E1-H1型猪GARV,可能为人-猪重配株,LLZ212基因片段VP7、VP4、NSP4和NSP5与人GARV同源性最高,LLZ212基因片段VP6、VP2、NSP2和NSP3猪GARV同源性最高。
A case of LARGEN G3 piglet GARV LLZ212 was amplified by PCR from VP7, VP4, VP6, VP2 and NSP2-5 in the viral diarrhea prevention and control center of China CDC, and its genotype was determined , And explored its evolutionary relationship with G3 type GARV in the locality. VP7, VP4, VP6, VP2 and NSP2-5 of LLZ212 and 6 G3 positive specimens from local children were amplified by reverse transcription-polymerase chain reaction (RT-PCR) , And sequenced. A group of rotavirus automatic typing tools (RotaC v2.0) for each fragment typing. Homology and phylogenetic analysis were performed using the DNAStar and MEGA 5.0 software package and the reference strain on GenBank. The genotype of LLZ212 in Lulong pig was G3-P [8] -I1-C1-N1-T1-E1-H1, -H1. The LLZ212 gene fragment VP7, VP4, NSP4 and NSP5 shared the highest homology with the human GARV E885, CMH014 / 07, Wa and RMC321 strains, respectively, and the phylogenetic tree was in the same sub-branch. The LL612 gene fragment VP6, PRG921 strain has the highest homology, VP2 gene has the highest homology with porcine GARV OSU strain, and the phylogenetic tree is in the same sub-branch. A rare G3-P [8] -I5-C1-N1-T1-E1-H1 pig GARV was identified as human-porcine, and LLZ212 gene fragments VP7, VP4, NSP4 and NSP5 shared homology with human GARV Highest, LLZ212 gene fragment VP6, VP2, NSP2 and NSP3 pig GARV homology highest.