中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2008年
5期
421-425
,共5页
吉彦莉%李凌君%韦献飞%王玲%常义斌%唐荣兰%朱永红%庄辉
吉彥莉%李凌君%韋獻飛%王玲%常義斌%唐榮蘭%硃永紅%莊輝
길언리%리릉군%위헌비%왕령%상의빈%당영란%주영홍%장휘
戊型肝炎病毒%猪%基因型%人畜共患病
戊型肝炎病毒%豬%基因型%人畜共患病
무형간염병독%저%기인형%인축공환병
Hepatitis E virus%Swine%Genotype%Zoonosis
目的 分析从广西地区分离的1株猪戊型肝炎病毒swGX32全基因组序列并比较其与其他分离株的差异.方法 设计PCR引物,用巢式反转录聚合酶链反应法(RT-nPCR)分段扩增戊型肝炎病毒(HEV)株swGX32全基因序列,用cDNA末端快速扩增法(RACE)扩增其末端序列,对扩增产物进行克隆和测序,并对拼接后的基因组进行序列和进化分析.结果 除3′polyA尾巴外,swGX32全长7240 nt, ORF1与ORF2重叠4 nt, ORF3包含在ORF2序列中.swGX32全基因序列与HEV1～4型核苷酸序列的同源性分别为:73%～74%、73%、74%～75%,83%～94%,其中与中国人源HEV株JKO-ChiSai98C同源性最高,达94%.全基因序列进化分析显示,swGX32位于HEV基因4型分枝上,ORF2部分核苷酸序列进化分析显示,swGX32与JKO-ChiSai98C同在HEV 4a亚型分枝上.结论 猪HEV swGX32在全基因组结构及分子进化上均与人HEV JKO-ChiSai98C有密切关系,为揭示戊型肝炎是一种人兽共患病提供了分子生物学依据.
目的 分析從廣西地區分離的1株豬戊型肝炎病毒swGX32全基因組序列併比較其與其他分離株的差異.方法 設計PCR引物,用巢式反轉錄聚閤酶鏈反應法(RT-nPCR)分段擴增戊型肝炎病毒(HEV)株swGX32全基因序列,用cDNA末耑快速擴增法(RACE)擴增其末耑序列,對擴增產物進行剋隆和測序,併對拼接後的基因組進行序列和進化分析.結果 除3′polyA尾巴外,swGX32全長7240 nt, ORF1與ORF2重疊4 nt, ORF3包含在ORF2序列中.swGX32全基因序列與HEV1～4型覈苷痠序列的同源性分彆為:73%～74%、73%、74%～75%,83%～94%,其中與中國人源HEV株JKO-ChiSai98C同源性最高,達94%.全基因序列進化分析顯示,swGX32位于HEV基因4型分枝上,ORF2部分覈苷痠序列進化分析顯示,swGX32與JKO-ChiSai98C同在HEV 4a亞型分枝上.結論 豬HEV swGX32在全基因組結構及分子進化上均與人HEV JKO-ChiSai98C有密切關繫,為揭示戊型肝炎是一種人獸共患病提供瞭分子生物學依據.
목적 분석종엄서지구분리적1주저무형간염병독swGX32전기인조서렬병비교기여기타분리주적차이.방법 설계PCR인물,용소식반전록취합매련반응법(RT-nPCR)분단확증무형간염병독(HEV)주swGX32전기인서렬,용cDNA말단쾌속확증법(RACE)확증기말단서렬,대확증산물진행극륭화측서,병대병접후적기인조진행서렬화진화분석.결과 제3′polyA미파외,swGX32전장7240 nt, ORF1여ORF2중첩4 nt, ORF3포함재ORF2서렬중.swGX32전기인서렬여HEV1～4형핵감산서렬적동원성분별위:73%～74%、73%、74%～75%,83%～94%,기중여중국인원HEV주JKO-ChiSai98C동원성최고,체94%.전기인서렬진화분석현시,swGX32위우HEV기인4형분지상,ORF2부분핵감산서렬진화분석현시,swGX32여JKO-ChiSai98C동재HEV 4a아형분지상.결론 저HEV swGX32재전기인조결구급분자진화상균여인HEV JKO-ChiSai98C유밀절관계,위게시무형간염시일충인수공환병제공료분자생물학의거.
Objective To analyze the complete genome sequence of Guangxi HEV isolate swGX32 and to compare it with other HEV isolates. Methods The overlapping fragments of HEV isolate swGX32 were amplified with reverse-transcription nested polymerase chain reaction (RT-nPCR),and the 5′ and 3′ ends of viral genome were amplified with rapid amplification of cDNA ends (RACE). The PCR products were cloned and sequenced. The sequence and phylogenetic analysis of swGX32 was performed. Results The genome of swGX32 consisted of 7240 nt excluding the polyA tail, with 4 nt overlapping between ORF1 and ORF2. ORF3 is contained in the sequence of ORF2. The complete genome sequence of swGX32 shared identity of 73%-74%, 73%, 74%-75%,83%-94% with HEV genotype 1,2,3 and 4, respectively. Among all these HEV reference sequences, swGX32 showed the highest identity with the human isolate JKO-ChiSai98C (94%). Phylogenetic tree showed that swGX32 belonged to genotype 4 and clustered with JKO-ChiSai98C in the branch of HEV subtype 4a. Conclusion The swine HEV isolate swGX32 is closely related to human strain JKO-ChiSai98C genetically and phylogenetically, which further provides molecular biology evidence of hepatitis E as a zoonosis.
