目的:探讨低水平HBV-DNA乙肝患者乙肝病毒外膜大蛋白(hepatitis B virus large surface protein,LHBs)的表达及其意义,从而探讨LHBs是否可以用作乙型肝炎e抗原(hepatitis B e antigen,HBeAg)阴性乙型肝炎患者病毒复制程度的判定指标。方...目的:探讨低水平HBV-DNA乙肝患者乙肝病毒外膜大蛋白(hepatitis B virus large surface protein,LHBs)的表达及其意义,从而探讨LHBs是否可以用作乙型肝炎e抗原(hepatitis B e antigen,HBeAg)阴性乙型肝炎患者病毒复制程度的判定指标。方法:对83例HBeAg阴性的慢性乙型肝炎患者血清,采用ELISA进行LHBs检测,全自动免疫分析仪检测HBeAg,实时荧光定量PCR方法检测HBV DNA。分析HBV DNA无拷贝数组、低拷贝数组及高拷贝数组中LHBs的检出率以及HBV DNA不同拷贝数组与LHBs的相关性。结果:在HBV DNA无拷贝数组,LHBs的阳性率为6.38%;低拷贝数组,LHBs的阳性率为56.25%,吸光度值与拷贝数对数值没有显著的相关性(r=0.25,P=0.36);高拷贝数组,阳性率为95%,吸光度值与拷贝数对数值有良好相关性(r=0.90,P<0.0001)。结论:检测HBeAg阴性慢性乙型肝炎患者血清LHBs,有助于判断患者体内HBV的复制程度,但是否能够作为HBeAg阴性乙型肝炎抗病毒治疗终点的有效判定指标,尚有待进一步探讨。展开更多
The N-terminal 54 base pairs (encoding amino acid residues 2—19) within the preS1 region of the human hepatitis B virus surface antigen gene were deleted by site-directed mutagenesis. Unlike the wild type large surfa...The N-terminal 54 base pairs (encoding amino acid residues 2—19) within the preS1 region of the human hepatitis B virus surface antigen gene were deleted by site-directed mutagenesis. Unlike the wild type large surface antigen protein; when this mutated gene was expressed in monkey kidney cell line COS-M6, the protein product (S301 protein) could be secreted from the cells. Moreover, the inhibition of the secretion of the major surface antigen protein by this altered large surface antigen protein was greatly reduced, suggesting that the deleted region contained a retention sequence which prevented the secretion of the large surface antigen. However, the coexpression of the major S protein was essential for the secretion of the S301 protein. When coexpressed, the secretion of these two proteins was synchronous. Like the wild type large surface antigen protein, the S301 protein could be translocated into ertdoplasmic reticulum and glycosylated after its synthesis in COS cells. The S301 protein was thermostable and proteinase-resistant. It also retained the antigenicity of the large S and major S proteins. Given the fact that the S301 protein is readily secretable, stable and identical to the large S protein in terms of their antigenicity, it may be developed into a new generation of recombinant vaccine for the prevention of viral hepatitis.展开更多
[目的]检测乙肝患者血清中乙肝表面抗原大蛋白（Hepatitis B Virus Large Surface Protein,LHBs）、乙肝前S1、HBV DNA,探讨LHBs用于乙肝患者临床诊断的意义。[方法]采用酶联免疫吸附实验（ELISA）检测LHBs以及乙肝前S1,采用化学发光方...[目的]检测乙肝患者血清中乙肝表面抗原大蛋白（Hepatitis B Virus Large Surface Protein,LHBs）、乙肝前S1、HBV DNA,探讨LHBs用于乙肝患者临床诊断的意义。[方法]采用酶联免疫吸附实验（ELISA）检测LHBs以及乙肝前S1,采用化学发光方法检测乙肝两对半,采用荧光定量PCR方法对患者HBV DNA进行检测。[结果]（1）相同乙肝模式患者血清LHBs与HBV DNA检出率比较差异无显著性意义（P〉0.05）;（2）HBeAg阳性患者血清中LHBs与HBV DNA阳性率均明显高于HBeAg阴性患者,差异有显著性意义（P〈0.05）;（3）相同乙肝模式患者血清乙肝前S1的检出低于HBV DNA的检出,两者相比较差异有显著性意义（P〈0.05）。[结论]LHBs可以弥补由于乙肝病毒变异引起的HBeAg检测的不足,LHBs可以反映病毒复制。展开更多
目的探讨乙型肝炎患者血清中乙型肝炎病毒外膜大蛋白（hepatitis B virus large surface protein，LHBs）与病毒复制及肝脏受损程度的相关性。方法随机选取武汉总医院2011年3月～6月乙肝患者血清标本192例，分别采用酶联免疫吸附实验（E...目的探讨乙型肝炎患者血清中乙型肝炎病毒外膜大蛋白（hepatitis B virus large surface protein，LHBs）与病毒复制及肝脏受损程度的相关性。方法随机选取武汉总医院2011年3月～6月乙肝患者血清标本192例，分别采用酶联免疫吸附实验（ELISA）检测LHBs，乙型肝炎病毒前S1抗原（PreS1-Ag），化学发光方法定量检测乙型肝炎病毒血清学模式，实时荧光定量PCR方法检测血清HBV-DNA，全自动生化分析仪检测血清谷氨酸氨基转移酶（ALT）。按乙型肝炎病毒e抗原阴阳性不同分组，比较LHBs，HBV-DNA及PreS1-Ag阳性率差异，同时分析LHBs含量与HBsAg，HBV-DNA及ALT水平的相关性。结果HBeAg阳性乙肝患者血清中，LHBs的阳性率与HBV-DNA阳性率差异无统计学意义（χ^2=0．32，P〉0．05），LHBs的阳性率与PreS1-Ag阳性率差异有统计学意义（χ^2=23．44，P〈0．05）；HBeAg阴性乙肝患者血清中，LHBs的阳性率与HBV-DNA和PreS1阳性率差异均有统计学意义（χ^2=11．73，28．55，P〈0．05）；LHBs含量与HBV—DNA拷贝数和表面抗原浓度呈正相关性（r=0．905，0．801，P〈0．05）；阳性组间LHBs含量与ALT指数无相关性。结论乙肝患者血清中LHBs的表达与HBV-DNA拷贝数、乙肝表面抗原浓度呈正相关，较PreS1-Ag更能反映乙肝患者病毒复制状况，尤其对HBeAg阴性患者抗病毒治疗及预后具有重要的指导意义，但不适用于判断肝细胞的受损程度。展开更多
文摘目的:探讨低水平HBV-DNA乙肝患者乙肝病毒外膜大蛋白(hepatitis B virus large surface protein,LHBs)的表达及其意义,从而探讨LHBs是否可以用作乙型肝炎e抗原(hepatitis B e antigen,HBeAg)阴性乙型肝炎患者病毒复制程度的判定指标。方法:对83例HBeAg阴性的慢性乙型肝炎患者血清,采用ELISA进行LHBs检测,全自动免疫分析仪检测HBeAg,实时荧光定量PCR方法检测HBV DNA。分析HBV DNA无拷贝数组、低拷贝数组及高拷贝数组中LHBs的检出率以及HBV DNA不同拷贝数组与LHBs的相关性。结果:在HBV DNA无拷贝数组,LHBs的阳性率为6.38%;低拷贝数组,LHBs的阳性率为56.25%,吸光度值与拷贝数对数值没有显著的相关性(r=0.25,P=0.36);高拷贝数组,阳性率为95%,吸光度值与拷贝数对数值有良好相关性(r=0.90,P<0.0001)。结论:检测HBeAg阴性慢性乙型肝炎患者血清LHBs,有助于判断患者体内HBV的复制程度,但是否能够作为HBeAg阴性乙型肝炎抗病毒治疗终点的有效判定指标,尚有待进一步探讨。
基金This research was supported in part by U. S. Public Health Service Grants CA-70175 and CA-2243 to J. Mertz from the U. S. National Institute of Health.
文摘The N-terminal 54 base pairs (encoding amino acid residues 2—19) within the preS1 region of the human hepatitis B virus surface antigen gene were deleted by site-directed mutagenesis. Unlike the wild type large surface antigen protein; when this mutated gene was expressed in monkey kidney cell line COS-M6, the protein product (S301 protein) could be secreted from the cells. Moreover, the inhibition of the secretion of the major surface antigen protein by this altered large surface antigen protein was greatly reduced, suggesting that the deleted region contained a retention sequence which prevented the secretion of the large surface antigen. However, the coexpression of the major S protein was essential for the secretion of the S301 protein. When coexpressed, the secretion of these two proteins was synchronous. Like the wild type large surface antigen protein, the S301 protein could be translocated into ertdoplasmic reticulum and glycosylated after its synthesis in COS cells. The S301 protein was thermostable and proteinase-resistant. It also retained the antigenicity of the large S and major S proteins. Given the fact that the S301 protein is readily secretable, stable and identical to the large S protein in terms of their antigenicity, it may be developed into a new generation of recombinant vaccine for the prevention of viral hepatitis.
文摘目的探讨乙型肝炎患者血清中乙型肝炎病毒外膜大蛋白（hepatitis B virus large surface protein，LHBs）与病毒复制及肝脏受损程度的相关性。方法随机选取武汉总医院2011年3月～6月乙肝患者血清标本192例，分别采用酶联免疫吸附实验（ELISA）检测LHBs，乙型肝炎病毒前S1抗原（PreS1-Ag），化学发光方法定量检测乙型肝炎病毒血清学模式，实时荧光定量PCR方法检测血清HBV-DNA，全自动生化分析仪检测血清谷氨酸氨基转移酶（ALT）。按乙型肝炎病毒e抗原阴阳性不同分组，比较LHBs，HBV-DNA及PreS1-Ag阳性率差异，同时分析LHBs含量与HBsAg，HBV-DNA及ALT水平的相关性。结果HBeAg阳性乙肝患者血清中，LHBs的阳性率与HBV-DNA阳性率差异无统计学意义（χ^2=0．32，P〉0．05），LHBs的阳性率与PreS1-Ag阳性率差异有统计学意义（χ^2=23．44，P〈0．05）；HBeAg阴性乙肝患者血清中，LHBs的阳性率与HBV-DNA和PreS1阳性率差异均有统计学意义（χ^2=11．73，28．55，P〈0．05）；LHBs含量与HBV—DNA拷贝数和表面抗原浓度呈正相关性（r=0．905，0．801，P〈0．05）；阳性组间LHBs含量与ALT指数无相关性。结论乙肝患者血清中LHBs的表达与HBV-DNA拷贝数、乙肝表面抗原浓度呈正相关，较PreS1-Ag更能反映乙肝患者病毒复制状况，尤其对HBeAg阴性患者抗病毒治疗及预后具有重要的指导意义，但不适用于判断肝细胞的受损程度。
