中华传染病杂志
中華傳染病雜誌
중화전염병잡지
CHINESE JOURNAL OF INFECTIOUS DISEASES
2011年
11期
679-684
,共6页
侯佩强%张荣强%边广%陆娟娟
侯珮彊%張榮彊%邊廣%陸娟娟
후패강%장영강%변엄%륙연연
流感病毒A型,H1N1亚型%基因,病毒%血凝素类,病毒%聚合酶链反应%序列分析%变异(遗传学)
流感病毒A型,H1N1亞型%基因,病毒%血凝素類,病毒%聚閤酶鏈反應%序列分析%變異(遺傳學)
류감병독A형,H1N1아형%기인,병독%혈응소류,병독%취합매련반응%서렬분석%변이(유전학)
Influenza A virus,H1N1 subtype%Genes,viral%Hemagglutinins,viral%Polymerase chain reaction%Sequence analysis%Variation (genetics)
目的 了解2009年度甲型H1N1流行性感冒(流感)病毒的检测情况和血凝素(HA)基因变异情况.方法 选择国家级流感监测哨点医院以及暴发疫情的疫点,采集流感样病例的鼻咽拭子标本,通过实时(RT)-PCR进行病毒分型及甲型H1N1流感病毒检测,对阳性标本采用狗肾细胞(MDCK)进行病原分离,采用红细胞凝集试验测定病毒效价,用血凝抑制实验进行型别鉴定,通过RT-PCR扩增毒株HA1片段的基因并进行序列测定,利用生物信息学技术进行序列分析.结果 共检测咽拭子样本996份,其中核酸检测阳性病例包括甲型H1N1 337份,季节性H1N1亚型1份,季节性H3N2亚型67份,B型12份,流感核酸检测阳性率为41.87％,其中甲型流感核酸检测阳性率为33.84％.分离出甲型H1N1病毒36株,选择18株.测序成功的10株甲型H1N1流感病毒在多个氨基酸位点发生变异,与疫苗株A/California/07/2009(H1N1)比较,有6个位点发生突变,其中1个位点位于抗原决定簇的B区.结论 2009年度分离到的流感病毒株中以甲型H1N1为绝对的优势毒株,毒株的血凝素基因与世界卫生组织(WHO)提供的疫苗株相比有变异,与疫苗株相比,抗原决定簇B区有改变,但关键位点第222位没有变化.
目的 瞭解2009年度甲型H1N1流行性感冒(流感)病毒的檢測情況和血凝素(HA)基因變異情況.方法 選擇國傢級流感鑑測哨點醫院以及暴髮疫情的疫點,採集流感樣病例的鼻嚥拭子標本,通過實時(RT)-PCR進行病毒分型及甲型H1N1流感病毒檢測,對暘性標本採用狗腎細胞(MDCK)進行病原分離,採用紅細胞凝集試驗測定病毒效價,用血凝抑製實驗進行型彆鑒定,通過RT-PCR擴增毒株HA1片段的基因併進行序列測定,利用生物信息學技術進行序列分析.結果 共檢測嚥拭子樣本996份,其中覈痠檢測暘性病例包括甲型H1N1 337份,季節性H1N1亞型1份,季節性H3N2亞型67份,B型12份,流感覈痠檢測暘性率為41.87％,其中甲型流感覈痠檢測暘性率為33.84％.分離齣甲型H1N1病毒36株,選擇18株.測序成功的10株甲型H1N1流感病毒在多箇氨基痠位點髮生變異,與疫苗株A/California/07/2009(H1N1)比較,有6箇位點髮生突變,其中1箇位點位于抗原決定簇的B區.結論 2009年度分離到的流感病毒株中以甲型H1N1為絕對的優勢毒株,毒株的血凝素基因與世界衛生組織(WHO)提供的疫苗株相比有變異,與疫苗株相比,抗原決定簇B區有改變,但關鍵位點第222位沒有變化.
목적 료해2009년도갑형H1N1류행성감모(류감)병독적검측정황화혈응소(HA)기인변이정황.방법 선택국가급류감감측초점의원이급폭발역정적역점,채집류감양병례적비인식자표본,통과실시(RT)-PCR진행병독분형급갑형H1N1류감병독검측,대양성표본채용구신세포(MDCK)진행병원분리,채용홍세포응집시험측정병독효개,용혈응억제실험진행형별감정,통과RT-PCR확증독주HA1편단적기인병진행서렬측정,이용생물신식학기술진행서렬분석.결과 공검측인식자양본996빈,기중핵산검측양성병례포괄갑형H1N1 337빈,계절성H1N1아형1빈,계절성H3N2아형67빈,B형12빈,류감핵산검측양성솔위41.87％,기중갑형류감핵산검측양성솔위33.84％.분리출갑형H1N1병독36주,선택18주.측서성공적10주갑형H1N1류감병독재다개안기산위점발생변이,여역묘주A/California/07/2009(H1N1)비교,유6개위점발생돌변,기중1개위점위우항원결정족적B구.결론 2009년도분리도적류감병독주중이갑형H1N1위절대적우세독주,독주적혈응소기인여세계위생조직(WHO)제공적역묘주상비유변이,여역묘주상비,항원결정족B구유개변,단관건위점제222위몰유변화.
Objective To understand the detections of influenza A (H1N1) in 2009,and haemagglutinin (HA) gene mutations and the comparisons with standard strains.Methods The nasopharyngeal swabs from patients with influenza-like illness (ILI) in National Influenza Sentinel Surveillance Hospital and the outbreak epidemic area were collected.The virus typing and A (H1N1) viruses were tested by real time-polymerase chain reaction (RT-PCR).Then the pathogens were isolated with MDCK cells,the virus titer was determined with hemagglutination test and the virus typing was identified with hemagglutination inhibition test (HA1).The RT-PCR products of HA1 gene of virulent strains were sequenced and then analyzed through bioinformatics.Results A total of 996 pharyngeal swab specimens were tested,and nucleic acid positive cases included 337 A (H1N1) subtype,1 seasonal A (H1N1) subtype,67 A (H3N2) subtype,and 12 B type.The positive rate of nucleic acid detection of influenza was 41.87％ and that of A (H1N1) was 33.84％.Thirty-six influenza A (H1N1) virus strains were isolated,and 10 of them were successfully sequenced and several amino acid mutations were identified.There were 6 amino acid mutations found compared with vaccine strain A/California/07/2009 (H1N1),and 1 site was in area B of epitope.Conclusions A (H1N1) is absolute predominant among isolated strains in 2009.HA gene of virulent strains is mutated compared with vaccine strain provided by World Health Organization,which shows that the area B of epitope changes,while the key amino acid position 222 doesn't change.
