酶联免疫法检测小儿乙肝PreS1抗原及抗体在乙肝诊断中的应用价值

目的探讨酶联免疫法检测小儿乙肝病毒前S1蛋白(PreS1)抗原及抗体在小儿乙肝诊断中的应用价值。方法在2014年至2016年在本院接受诊断和治疗的乙肝患儿中,选择200例资料完整且接受两对半检查和酶联免疫法检测血清PreS1抗原及抗体的患儿作为研究对象,计算不同两对半检测结果患儿PreS1抗原及抗体阳性率,分析PreS1抗原阳性与HBV-M的Spearman相关性。结果 200例乙肝患者入选。18例乙肝大三阳患儿的PreS1抗原阳性及抗体阳性的检出率较高,均为83.33%。25例PreS1抗原阳性组患儿与175例抗原阴性组患儿HBsAg、HBeAg和HBcAb数据对比差异具有统计学意义,说明PreS1抗原阳性与HBsAg、HBeAg和HBcAb具有正相关关系,相关数值分别为0.620、0.550、0.720,P值分别为0.021、0.009、0.020。结论乙肝PreS1抗原及抗体阳性检测准确率相对较高,可以作为乙肝诊断的补充检验,为患儿的临床诊断和治疗提供依据。

酶联免疫法检测乙肝PreS1抗原及抗体在乙肝诊断中的应用价值

目的 探究酶联免疫法检测乙肝PreS1抗原及抗体在乙肝诊断中的应用价值.方法 选择2014年8月-2016年8月乙肝＂两对半＂检查结果中几种常见模式为样本,采用乙肝＂两对半＂7种常用检测模式对140例行PreS1抗原、抗体检测.经酶联免疫法检测PreS1抗原及抗体.结果 在大三阳患者中PreS1抗原阳性87.5%,在小三阳患者中PreS1抗原阳性8.6%,在HBsAg、HBcAb阳性患者中PreS1抗原阳性13.3%,在单纯HBsAg阳性患者中PreS1抗原阳性4.3%.HBV-DNA总阳性率与PreS1总阳性率对比差异无统计学意义,且两者总符合率较高.PreS1抗原阳性与HBsAg、HBeAg及HBcAb阳性相关（P〈0.05）;与HBsAb、HBeAb阳性无关.结论 经乙肝PreS1抗原及抗体检测,可有效补充乙肝诊断,及早发现已存在的乙肝感染.

乙肝病毒PreS1抗原和乙肝两对半结合检查的意义和应用

目的:评价检测乙肝病毒PreS1抗原与乙肝两对半相结合检查的临床应用及在健康体检中的价值。方法:将临床送检的580份血清标本用ELASE进行HBV测定,同时进行PreS1抗原检测。结果:PreS1总阳性率为33%,HBeAg阳性率为24.2%。PreS1总阳性率明显高于HBeAg阳性率。结论:PreS1抗原作为乙肝感染和复制的血清学指标敏感性与HBeAg具有一致性和相符性,对提示乙肝患者的病情发展和转归有重要临床意义。PreS1可以作为一项新的乙肝病毒复制的传染性指标。

乙型肝炎病毒血清标志物与PreS1抗原的关系

目的:了解PreS1与HBVM和HBV DNA的关系及其临床应用价值。方法:对306例临床血清采用ELISA方法检测HBVM和PreS1,荧光定量PCR法检测HBV DNA。结果:中HBeAg阳性146例,其中前S1阳性127例阳性率88.1%,HBeAg阴性共160例,其中前s72例阳性率45%,HBeAg阴性血清160例前s多数存在于抗HBe阳性血清中,阳性率40.6%。结论:PreS1比HBeAg更能较好反映病毒存在和复制状态.HBVM、PreS1和HBV DNA联合检测,更有利于乙肝病情监测和疗效评估。

乙型肝炎病毒PreS1抗原与HBV血清标志物的相关性

传统乙肝血清标志物是目前我国用于乙肝检测与筛查的普及指标，但其在检测病毒复制等方面尚有不足。文章分析前Sl抗原（PreSl）和乙型肝炎病毒（HBV）血清标志物之间的关系，探讨乙型肝炎病毒前S1蛋白在临床上的应用价值。关于PreSl、抗PreSl与HBV免疫标志物的相关性。采用ELISA对306例HBsAg阳性标本进行PreSl、抗PreSl与HBV血清标志物相关性的研究证实，前S1抗原（Pre—SlAg）检测是对乙肝“两对半”，尤其是e抗原和HBV—DNA测定的重要补充和加强。

Detection of anti-preS1 antibodies for recovery of hepatitis B patients by immunoassay

AIM: To establish a convenient immunoassay method based on recombinant antigen preS1(21-119aa) to detect anti-preS1 antibodies and evaluate the clinical significance of antibodies in hepatitis B. METHODS: The expression plasmid pET-28a-preS1 was constructed, and a large quantity of preS1(21-119aa) fragment of the large HBsAg protein was obtained. The preS1 fragment purified by Ni(2+)-IDA affinity chromatography was used as coated antigen to establish the indirect ELISA based on streptavidin-biotin system for detection of the anti-preS1 antibodies in sera from HBV-infected patients. For follow-up study, serial sera were collected during the clinical course of 21 HBV-infected patients and anti-preS1 antibodies, preS1 antigen, HBV-DNA and other serological HBV markers were analyzed. RESULTS: preS1(21-119aa) fragment was highly expressed from the plasmid pET-28a-preS1 in a soluble form in E.Coli (30mg.L(-1)), and easily purified to high purity over 90% by one step of Ni(2+)-IDA-sepharose 6B affinity chromatography. The purity and antigenicity of the purified preS1(21-119aa) protein was determined by 150g.L(-1) SDS-PAGE, Western blot and a direct ELISA. Recombinant preS1(21-119aa) protein was successfully applied in the immunoassay which could sensitively detect the anti-preS1 antibodies in serum specimens of acute or chronic hepatitis B patients. Results showed that more than half of 19 acute hepatitis B patients produced anti-preS1 antibodies during recovery of the disease, however, the response was only found in a few of chronic patients. In the clinical follow-up study of 11 patients with anti-preS1 positive serological profile, HBsAg and HBV-DNA clearance occurred in 6 of 10 acute hepatitis B patients in 5-6 months, and seroconversion of HBeAg and disappearance of HBV-DNA occurred in 1 chronic patients treated with lavumidine, a antiviral agent. CONCLUSION: The high-purity preS1(21-119aa) coated antigen was successfully prepared by gene expression and affinity chromatography. Using this antigen, a conveniently detective system of anti-preS1 antibodies in sera was established. Preliminarily clinical trial the occurrence of anti-preS1 antibodies in acute hepatitis B patients suggests the clearance of HBV from serum in a short-term time, and anti-preS1 positive in chronic patients means health improvement or recovery from the disease.

Identification of the immunogenic domains in HBsAg preS1 region using overlapping preS1 fragment fusion proteins

AIM: The incorporation of hepatitis B virus (HBV) preS1 region into epitope-based vaccines against HBV has been accepted widely, but the incorporate site and size of preS1 sequence is controversial. Therefore our purpose was to further investigate its immunogenic domains for the epitopebased hepatitis B vaccine design.METHODS: Eight GST fusion proteins containing overlapping preS1 fragments in preS1 (21-119) region were expressed in E.coli. Using these purified fusion proteins, the immunogenic domains in preS1 region were identified in detail in mice and humans by Western blot analysis and ELISA.RESULTS: The results in mice showed that the immunogenic domains mainly existed in preS1 (21-59) and preS1 (95-109). Similarly, these fragments had strong immunogenicity in humans; whereas the other parts except for preS1 (60-70) also had some immunogenicity.More importantly, a major immunogenic domain, preS1 (34-59), which has much stronger immunogenicity, was identified. Additionally, the antibodies against some preS1 fragments, especially preS1 (34-59), were speculated to be virus-neutralizing.CONCLUSION: Eight GST fusion proteins containing overlapping preS1 fragments were prepared successfully. They were used for the study on the immunogenic domains in preS1 (21-119) region. The preS1 (34-59) fragments were the major immunogenic domains in the preS1 region, and the antibodies against these fragments were speculated to be virus-neutralizing. Therefore, the incorporation of preS1 (34-59) fragments into epitopebased HBV vaccines may be efficient for enhancement of immune response. Additionally, the results also imply that there are more complex immune responses to preS1 region and more abundant immunogenic domains in humans.

THE COEXPRESSION OF THE preS1(1-42) AND THE CORE(1-144)ANTIGEN OF HBV IN E.coli

Objective.To study the therapeutic T cell vaccine for the treatment of chronic hepatitis B by improving the cellular immunization of HBsAg vaccine with the coexpression of the preS1(1-42)and the Core(1-144)anti-gen of HBV in E.coli.Methods.The genes of HBcAg (1-144)and preS1(1-42)were amplified and fused by PCR.This fused gene was inserted in the prokaryotic expression vector pET-11d and expre ssed in E.coli.Results.It was showed by SDS-PAGE that the pr otein molecular weight of the coexpr ession product was about 20kD,20%of all bacteria prote in.The monoclonal antibodies again st core and preS1antibody could re-act with this fused protein by Western-blot technique respectively.The fused gene was verified by sequencin g.Under the immune electron microscop y,this fused protein is typical particles of HBcAg but in an aggregated form.Conclusion.The results might aid for studying T c ell immunotherapeutic vaccine for c hronic hepatitis B.

乙肝病毒前S_1抗原与乙肝两对半检测的相关性及其临床价值

目的对乙肝两对半检测HBSAg阳性患者进行乙肝病毒前S1抗原(PreS1)检测并对比分析,进而体现乙肝病毒PreS1检测的临床价值。方法采用酶联免疫法(ELISA)对176例乙肝HBsAg阳性患者PreS1和两对半进行检测,比较各组PreS1及乙肝两对半结果,并结合临床进行分析。结果与结论血清乙肝病毒PreS1测定可以完善和补充乙肝两对半的检测,对HBV感染、复制及预后有着重要的临床意义。

乙肝前S1抗原临床应用价值的探讨

目的:通过乙肝前S1抗原与乙肝DNA和乙肝两对半的相关性分析,进一步探明PreS1-Ag对乙肝的诊断价值和其临床意义。方法:用酶联免疫吸附法检测PreS1-Ag和乙肝两对半,荧光定量PCR法检测HBV-DNA。结果:PreS1-Ag阳性率为61.5%(96/156),HBV-DNA阳性率为57.8%(90/156),HBeAg阳性率为32.1%(50/156)。结论:PreS1-Ag是反映HBV感染、复制的标志之一,而且比HBeAg敏感性更高,有很好的独立参考价值,并对乙肝两对半、HBV-DNA的检测有重要的补充作用。

乙肝病毒前S1抗原、乙肝病毒血清标志物和HBV-DNA联合检测的临床应用

目的探讨乙肝病毒前S1抗原(PreS1)、乙肝病毒标志物(HBVM)与乙肝病毒DNA(HBV-DNA)检测三者之间的关系,判断乙肝病毒在体内的复制情况。方法对550例乙型肝炎患者血清,用ELISA方法检测PreS1、两对半标志物,用荧光定量PCR方法定量检测HBV-DNA并分析3者之间的关系。结果HBsAg(+)、HBeAg(+)、HBcAb(+)的模式中,PreS1与HBV-DNA检出率分别为52.3%与88.4%,HBsAg(+)、HbeAb(+)、HBcAb(+)的模式中PreS1和HBV-DNA检出率分别为31.1%和47.5%,HBsAg(+)、HBcAb(+)的模式中PreS1和HBV-DNA检出率分别为47.3%和63.4%。结论PreS1能较好的反映HBV病毒感染及复制情况,可作为一种新的HBV血清标志物和监测指标,三者联合检测互相补充,更有助于临床疾病的诊治。

术前筛查HBeAg阴性HBV感染者检测乙肝大蛋白的意义

目的了解肝功能正常、HBeAg阴性HBV感染的术前筛查患者乙肝大蛋白(HBV-LP)阳性情况,评判血清HBV-LP检测在这类乙型肝炎患者中的意义。方法使用ELISA法检测180例HBeAg阴性HBV感染的术前筛查患者HBV-LP、乙肝前S1(HBVpreS1),同时采用实时定量PCR法做HBVDNA水平检测。结果 180份HBeAg阴性HBV感染血清PreS1-Ag、HBV-LP和HBVDNA检测阳性率分别为21.1%、34.4%和32.7%,χ2检验显示三组检测阳性率差异有统计学意义(χ2=9.1457,P=0.0103),用Scheffe法做率的两两比较显示:PreS1-Ag检测阳性率明显低于HBV-LP和HBVDNA(P<0.05),而HBV-LP和HBVDNA检测阳性率差异无统计学意义(P>0.05)。HBV-LP和HBVDNA配对检测显示结果差异无统计学意义(χ2=0.1739,P=0.6767),同时相关性分析示良好相关性(χ2=95.655,P=0.0000,r=0.7217)。49例HBVDNAcopies/ml常用对数值与HBV-LP定量OD值的散点图显示正相关(r=0.6192)。结论对术前筛查HBeAg阴性乙型肝炎携带者,HBV-LP在反应病毒复制方面是一个优于HBVpreS1检测的良好指标,与HBVDNA检测有良好的相关性,在不能开展HBVDNA检测的基层单位可作为评价乙型肝炎传染性的一种替代与补充。

前S1蛋白(PreS1)在乙型肝炎诊断中的作用

目的探讨HBV感染者血清中PreS1与HBV-DNA、HBeAg三者间的关系,进而评估PreS1在HBV感染、复制、诊断中的作用。方法收集1750例乙型肝炎患者血清,按不同HBV-M进行分组:HBsAg(+)、HBeAg(+)、HBcAb(+)为A组,HBsAg(+)、HBeAb(+)、HBcAb(+)为B组,HBsAg(+)、HBcAb(+)为C组,HBsAg(+)为D组,采用酶联免疫法(ELISA)检测乙肝病毒PreS1与HBV血清标志物,荧光定量PCR法检测HBV-DNA。结果以HBV-DNA>5×102拷贝/mL为阳性诊断标准,不同HBV-M模式中,PreS1与HBV-DNA的检出率无显著差异(P>0.01),PreS1与HBV-DNA的总符合率高于HBeAg与HBV-DNA的符合率,在HBeAg(-)患者中仍可不同程度地检出PreS1与HBV-DNA。结论PreS1可作为HBV感染、复制的一项新指标,在乙型肝炎诊断中具有很大的应用价值。

乙肝病毒前S1抗原时间分辨荧光免疫分析试剂盒的研制

目的利用时间分辨荧光免疫分析(TRFIA)技术建立乙型肝炎病毒前S1抗原的检测试剂盒。方法应用双抗体夹心法建立乙肝preS1抗原TRFIA检测试剂盒,对试剂盒的各项性能指标进行评估。结果对280份血清样本进行检测并与国产酶联免疫法试剂盒对比,结果显示自制试剂盒的特异性更好;200份正常人血清样本检测结果表明,该试剂盒的cutoff值=阴性对照荧光值×4.5。用自制质控品检测分析内和分析间的精密度分别为3.4%～7.8%,6.9%～8.4%;检测HBsAg及HBeAg无交叉反应。结论试剂盒各项指标均达到临床检测要求,TRFIA法精密度和特异性有较大提高,可替代酶免法试剂盒。

乙肝HBV DNA荧光定量检测与乙肝五项及perS1的相关性分析

目的分析乙肝五项及PreS1-Ag联合检测与HBV-DNA结果的一致性,探讨三者的相关性。方法分别采用ELISA、PCR的方法对328例乙肝患者进行HBV五项、前S1抗原及HBV-DNA的检测。结果在HBsAg(+)HBeAg(+)HBcAb(+)的模式下,HBV-DNA、PreS1-Ag的阳性率分别为81.0%、79.4%,在HBsAg(+)HBeAg(+)的模式下,HBV-DNA、PreS1-Ag的阳性率分别为85.7%、80.0%,两者的检出率差异无统计学意义(P>0.05);在HBsAg(+)HBeAb(+)HBcAb(+)模式下,HBV-DNA、PreS1-Ag的阳性率分别为40.8%、35.7%,在HBsAg(+)HBcAb(+)模式下,HBV-DNA、PreS1-Ag的阳性率分别为54.5%、48.5%,在HBsAg(+)HBeAb(+)模式下,HBV-DNA、PreS1-Ag的阳性率分别为50.0%、50.0%,在HBsAg(+)模式下,HBV-DNA、PreS1-Ag的阳性率分别为100%、100%。PreS1-Ag和HBeAg的阳性检出率在HBV-DNA阳性与阴性标本中差异均有统计学意义(P<0.005)。结论 PreS1-Ag、乙肝五项及HBV-DNA的结果密切关联,联合检测PreS1-Ag、乙肝五项能够很好的反映乙肝病毒的复制及传染性,对于乙肝的诊治具有指导意义。