中华肝脏病杂志
中華肝髒病雜誌
중화간장병잡지
CHINESE JOURNAL OF HEPATOLOGY
2012年
9期
654-658
,共5页
王琦%杨春%苏建家%曹骥%欧超%杨芳%张晶晶%史後林%杨芳%汪多平%王晓娟%王佳%阮萍%李瑗
王琦%楊春%囌建傢%曹驥%歐超%楊芳%張晶晶%史後林%楊芳%汪多平%王曉娟%王佳%阮萍%李瑗
왕기%양춘%소건가%조기%구초%양방%장정정%사후림%양방%왕다평%왕효연%왕가%원평%리원
肝炎病毒,乙型%模型,动物%树鼩
肝炎病毒,乙型%模型,動物%樹鼩
간염병독,을형%모형,동물%수구
Hepatitis B virus%Models,animal%Tree shrews
目的 探索影响树鼩体内长期感染HBV的因素.方法 77只新生期和49只非新生期树鼩,分别以不同的感染源和注射方式接种HBV,部分动物加以辅助措施.接种后定期获取动物血和肝组织,应用荧光定量聚合酶链反应、酶联免疫吸附试验、时间分辨免疫荧光分析、Southern blot和dotblot、免疫组织化学染色、电子和光学显微镜等多种方法,长期动态观察动物体内HBV感染标志的消长.结果 新生期接种的树鼩中6只确定、7只疑似为持续感染,非新生期接种的树鼩无一为持续感染.单份的HBV患者或树鼩血清为效果较好的感染源；皮下注射为较好的接种途径；接种用血清先经纯化有一定的辅助作用；联合检测树鼩血清的HBV免疫学标志和肝组织的HBV DNA为较好的检测方案.结论 不同的接种年龄、感染源、接种方法、辅助措施可影响树鼩体内慢性感染HBV的效果,合适的检测方案有助于判断和预测树鼩的感染状态.
目的 探索影響樹鼩體內長期感染HBV的因素.方法 77隻新生期和49隻非新生期樹鼩,分彆以不同的感染源和註射方式接種HBV,部分動物加以輔助措施.接種後定期穫取動物血和肝組織,應用熒光定量聚閤酶鏈反應、酶聯免疫吸附試驗、時間分辨免疫熒光分析、Southern blot和dotblot、免疫組織化學染色、電子和光學顯微鏡等多種方法,長期動態觀察動物體內HBV感染標誌的消長.結果 新生期接種的樹鼩中6隻確定、7隻疑似為持續感染,非新生期接種的樹鼩無一為持續感染.單份的HBV患者或樹鼩血清為效果較好的感染源；皮下註射為較好的接種途徑；接種用血清先經純化有一定的輔助作用；聯閤檢測樹鼩血清的HBV免疫學標誌和肝組織的HBV DNA為較好的檢測方案.結論 不同的接種年齡、感染源、接種方法、輔助措施可影響樹鼩體內慢性感染HBV的效果,閤適的檢測方案有助于判斷和預測樹鼩的感染狀態.
목적 탐색영향수구체내장기감염HBV적인소.방법 77지신생기화49지비신생기수구,분별이불동적감염원화주사방식접충HBV,부분동물가이보조조시.접충후정기획취동물혈화간조직,응용형광정량취합매련반응、매련면역흡부시험、시간분변면역형광분석、Southern blot화dotblot、면역조직화학염색、전자화광학현미경등다충방법,장기동태관찰동물체내HBV감염표지적소장.결과 신생기접충적수구중6지학정、7지의사위지속감염,비신생기접충적수구무일위지속감염.단빈적HBV환자혹수구혈청위효과교호적감염원；피하주사위교호적접충도경；접충용혈청선경순화유일정적보조작용；연합검측수구혈청적HBV면역학표지화간조직적HBV DNA위교호적검측방안.결론 불동적접충년령、감염원、접충방법、보조조시가영향수구체내만성감염HBV적효과,합괄적검측방안유조우판단화예측수구적감염상태.
Objective To determine the methods for establishing an in vivo model of long-term hepatitis B virus (HBV) infection in the Chinese tree shrew (Tupaia belangeri chinensis).Methods Seventyseven neonate (1-3 days old) and 49 young adult (2 weeks to 1 year old) tree shrews were inoculated with different HBV sources (chronic hepatitis B (CHB) human patient serum,single or pooled; HBV-infected tree shrew serum,single only; HBV-infected HepG2.2.15 cells' culture medium supernatant; HBV genome-transfected HepG2.2.15 cells' culture medium supernatant) through various routes of injection (subcutaneous,intraperitoneal,and direct liver via abdominal skin; adults also received intravenous and indirect liver via spleen).Serum and liver biopsies were collected from the animals at various time points post-inoculation for detection of HBV markers by fluorescence quantitative polymerase chain reaction,enzyme-linked immunosorbent assay,time-resolved immunofluorescence,Southern blotting,dot blotting,immunohistochemistry,and microscopy.Results Among the neonatal group of tree shrews,six (7.8％) were confirmed as HBV-infected for more than 72 (up to 228) weeks after inoculation and another seven (9.1％) were suspected of persistent infections.None of the young adult tree shrews developed persistent infection.Inoculation with single-source serum from either CHB humans or tree shrews were responsible for the most cases of infections,and the subcutaneous injection produced more infections than the other inoculation routes.The most reliable methods of determining HBV infection status were detection of serum HBV immunoreactive markers and intrahepatic HBV DNA.Conclusion In order to establish an in vivo model of CHB in the tree shrew,the animals should be inoculated in the neonatal period using subcutaneous injection.
