戊型肝炎病毒核酸阳性血浆经输血传播感染恒河猴的研究

目的 了解戊型肝炎病毒(HEV)核酸阳性血浆对灵长类动物的感染性和致病性。 方法 对抗-HEV IgM阳性而IgG阴性志愿献血员血浆进行HEV RNA检测,并将存在病毒血症献血员的10ml血浆静脉输入健康恒河猴,观察其对恒河猴的感染性和致病性。 结果 从1份抗-HEV IgM阳性而IgG阴性志愿献血员血浆中分离出HEV基因IV型RNA片段。该份血浆输入恒河猴后,恒河猴出现典型急性肝炎生物化学和病理表现,病毒血症,血清抗-HEV IgM和IgG抗体阳转。 结论 HEV病毒血症献血员血浆输入可以引起灵长类动物的HEV感染以及急性肝炎,提示HEV经输血传播的可能性

戊型肝炎病毒衣壳蛋白中和表位间的构象诱导

重组蛋白NE2包含了戊型肝炎病毒(HEV)衣壳蛋白(pORF2)的aa394～606片段。在NE2上已鉴定出了2个HEV中和表位,并获得了3个识别中和表位的单克隆抗体(MAb)8C11、13D8和8H3。这3个MAb间的交叉阻断ELISA实验发现,8C11和13D8可以彼此完全阻断,8H3对8C11和13D8均不能阻断,而8C11非但不能阻断8H3,反而显著增强了8H3与抗原的结合。用生物传感器进行的抗体与抗原结合的动力学分析也证实了这一现象。这些结果提示,在NE2上8H3表位区域受到抗原上某些结构的掩盖,而8C11与NE2的结合引起了抗原空间结构的改变,导致了掩盖8H3表位的结构的去除和8H3表位的充分暴露。免疫捕获RT PCR发现,8C11同样可以显著增强8H3对天然HEV病毒的捕获能力,提示这种结合诱导的衣壳蛋白空间构象改变在天然HEV病毒颗粒上同样存在

集美大学2002级福建籍新生肝炎感染现状

目的　了解集美大学 2 0 0 2级福建籍新生甲、乙、戊 3型肝炎感染情况。方法　采用血清流行病学调查方法。结果　学生中抗HAV -IgG ,HBsAg和抗HEV -IgG阳性率分别为 77.0 8% ,1 5 .2 3 % ,1 7.1 5 % ,地区差异明显 ;HAV ,HEV无性别差异 ,而HBV以男性为高 ;农村HAV和HBV感染率均高于城镇 ,而HEV感染率无城乡差别。结论　应该加强对本地区大学生的甲、乙肝预防 ,急需研究出一种有效的戊肝疫

Preparation and application of monoclonal antibodies against Herpes simplex virus-1

目的:制备并筛选HSV-1单抗,建立定量检测HSV-1病毒颗粒抗原的双抗体夹心ElISA方法,用于HSV-1病毒颗粒的质控。方法:以HSV-1免疫bAlb/C小鼠制备单克隆抗体,以筛选的中和单克隆抗体1f6为捕捉抗体,HrP标记的2b1为检测抗体,构建定量检测HSV-1病毒颗粒抗原的双抗体夹心ElISA方法,并对本方法的特异性、灵敏度、精密度、准确性和线性等性能进行验证。用本方法定量检测的病毒量与病毒滴度作回归分析。结果:构建的双抗体夹心定量检测HSV-1病毒颗粒抗原的ElISA方法,线性范围为0.125~2μg/Ml,相关系数为r2=0.995 5,定量限度为0.125μg/Ml,试剂的变异系数CV<10%,抗原回收率介于85.6%~107.1%之间。与HSV-1以外的其他样本无交叉反应。本方法检测与病毒感染滴度具有很好的相关性。结论:成功构建了定量检测HSV-1含量的ElISA方法,为HSV-1病毒颗粒抗原定量检测提供快速手段。Objective: To prepare and screen monoclonal antibodies against Herpes simplex virus-1( HSV-1),and develop a double antibody sandwich quantitative enzyme-linked immunosorbent assay( Q-ELISA) for detection of HSV-1 particle.This method was used to control the quality of viral particle in the developing and manufacturing process of HSV-1.Methods: BALB / c mice was immunized with HSV-1 to prepare monoclonal antibodies.A double antibody sandwich Q-ELISA was developed to determine concentration of HSV-1 particle,which was based on the neutralizing monoclonal antibody 1F6 as capture antibody,and 2B1 as HRP-conjugated antibody.The performance of the reagent was evaluated,including specificity,sensitivity,precision,accuracy and linear.And the relation between the amount of virus detected by this method and the virus titer was analyzed by regression analysis method.Results: The QELISA for HSV-1 particle was developed.The quantitation scope was 0.125- 2 μg / ml,the coefficient correlation was 0.995 5,the limit of detection was 0.125 μg / ml,the recovery was between 85.6% and 107.1%,the variation coefficient was lower than 10%,and the reagent does not react with other samples except HSV-1 antigen.This method has a good correlation with virus titer.Conclusion: The QELISA for HSV-1 particle was successfully developed,which provide a new approach for rapid and quantitative detection of HSV-1 antigen

Development of a quantitative ELISA detection method for Coxsackievirus A group 16 strain(CA16) antigen

目的:建立柯萨奇病毒A组16型(CA16)抗原的双抗体夹心ElISA定量检测方法,用于CA16灭活疫苗的研发和生产过程的抗原定量检测。方法:以CA16中和单抗T26H12为包被抗体、nA14b9为标酶抗体,构建定量检测CA16抗原的双抗体夹心ElISA方法,并对方法的特异性、灵敏度、精密度、准确性、线性和稳定性进行分析。结果:建立了双抗体夹心定量检测CA16抗原的ElISA方法。方法的线性相关系数r2=0.998,线性范围为8~128 ng/Ml,定量限度为8 ng/Ml;变异系数CV80%;与CA16以外的其他样本没有交叉反应。结论:构建的CA16抗原ElISA定量检测方法的各项性能符合定量检测需要,可用于CA16疫苗的研发和生产过程的抗原活性的定量检测。Objective:To develop an a quantitative enzyme linked immunosorbent assay(Q-ELISA) to determine the concentration of Coxsackievirus A Group 16 Strain(CA16) antigen.This method was used to determine CA16 antigen content at each stage of CA16 vaccine developing and manufacturing process.Methods:A double antibody sandwich Q-ELISA was developed to determine concentration of CA16 antigen,which was based on the high-affinity neutralizing monoclonal antibodies T26H12 as capture antibodies,and NA14B9 as HRP-labeled antibody.The performance of reagent were evaluated.Results:The Q-ELISA for CA16 antigen content was successfully developed.The reagent had good performance.The quantitation scope was 8-128 ng/ml,the coefficient correlation was 0.998,the limit of detection was 8 ng/ml,the recovery was between 87% and 113.8%.The stability was up to 80% after reagent was heated for 6 days at 37℃.The variation coefficient was lower than 15%,and thereagent was no reaction with other sample except CA16 antigen.Conclusion:The Q-ELISA for CA16 antigen was developed with good specificity,accuracy,precision and stability.The method can be used to determine CA16 antigen content during development and production of CA16 vaccine

Development of aquantitative ELISA detection method for Varicella Zoster Virus(VZV) antigen

目的:建立水痘-带状疱疹病毒(VzV)抗原的双抗体夹心ElISA定量检测方法,用于质控VzV灭活疫苗研发和生产中抗原含量。方法:以VzV中和单抗5f6C8为包被抗体、8H5d1为酶标抗体,构建定量检测VzV抗原的双抗体夹心ElISA方法,并对本方法的特异性、灵敏度、准确性、线性和稳定性等性能进行分析。结果:建立的双抗体夹心定量检测VzV抗原的ElISA方法,线性范围为0.4μg~13μg/Ml,相关系数为r2=0.994,定量限度为0.4μg/Ml;变异系数CV80%。与VzV以外的相关病毒样本没有交叉反应。结论:构建的VzV抗原ElISA定量检测方法的各项性能符合定量检测需要,可用于VzV灭活疫苗的研发和生产过程的抗原含量检测。Objective:To develop a quantitative enzyme linked immunosorbent assay(Q-ELISA) to determine the concentration of Varicella Zoster Virus(VZV) antigen.This method was used to determine VZV antigen content at each stage of VZV inactived vaccine developing and manufacturing process.Methods: A double antibody sandwich Q-ELISA was developed to determine concentration of VZV antigen,which was based on the the high-affinity neutralizing monoclonal antibodies 5F6C8 as capture antibodies,and 8H5D1 as HRP-labeled antibody.The performance of reagent were evaluated.Results: The Q-ELISA for VZV antigen content was successfully developed.The reagent had good performance.The quantitation scope was 0.4 μg~13 μg/ml,The coefficient correlation was 0.994,the limit of detection was 0.4 μg /ml,the recovery was between 87.5% and 111.6%.The stability was up to 80% after reagent was heated for 6 days at 37℃.The variation coefficient was lower than 15%,and the reagent was no reaction with other sample except VZV antigen.Conclusion: The Q-ELISA for VZV antigen was developed with good specificity,accuracy and stability.The method can be used to determine VZV antigen content during development and production of VZV inactived vaccine

Screening of adjuvant enhancing cellular immune response induced by ESAT6-CFP10 fusion protein in mice

通讯作者：叶祥忠，E-mail：[email protected][中文文摘] 目的 筛选能增强特异性抗原早期分泌抗原靶6蛋白(Early secretory antigenic target6,ESAT6)-培养滤出液蛋白-10(Culture filtrate protein10,CFP-10)融合蛋白(E1C0)诱导小鼠细胞免疫应答的佐剂,建立基于细胞免疫应答的小鼠模型,以评价基于体外干扰素γ释放分析(IFNγrelease assay,IGRA)结核诊断方法中特异性刺激抗原E1C0的活性。方法 建立小鼠IFNγ双抗体夹心SABC-ELISA检测系统,并验证系统的线性、灵敏度、重复性和特异性。将BALB/c小鼠随机分为7组:E1C0+单磷酸类脂A(Monophosphoryl lipid A,MPL)+双十八烷基二甲基溴化铵(Dimethyl dioctadecylammonium bromide,DDA)组、E1C0+DDA组、E1C0+MPL组、E1C0+弗氏不完全佐剂(IFA)组、E1C0组、生理盐水组和MPL+DDA联合组,每组6只,经小鼠后肢内侧皮下免疫3次,间隔2周,免疫剂量为:E1C0100滋g/只,MPL25μg/只,DDA250μg/只,IFA100滋l/只。末次免疫4周后处死小鼠,无菌取脾,分离脾淋巴细胞,加入E1C0进行培养,MTT法检测特异性淋巴细胞增殖反应,ELISA法检测培养上清中IFNγ水平。采用筛选出的最佳佐剂与抗原组合免疫3批BALB/c小鼠,进行IFNγ诱生测定。结果 检测系统的线性范围为:40～2560pg/ml(R>0.98);灵敏度为40pg/ml;变异系数(CV)0.05)。结论 E1C0与MPL和DDA联合免疫所诱导的小鼠Th1型细胞免疫应答最强,成功建立了用于评价刺激抗原E1C0活性的小鼠模型。[英文文摘]Objective To screen the adjuvant enhancing the cellular immune response induced by early secretory antigenic target 6（ESAT6）-culture filtrate protein-10（CFP10）in mice, and establish an animal model based on cellular immunγe response for evaluation of activity of specific stimulating antigen E1C0 in IFNγ release assay（IGRA）for diagnosis of tuberculosis（TB）. Methods mDouble antibody sandwich SABC-ELISA system for mouse IFNγ was developed and verified for linearity, sensitivity, reproducibility and specificity. BALB/c mice were randomly divided into seven groups, 6 for each, and immunized s.c. with E1C0 + monophosphoryl lipid A（MPL）+ dimethyl dioctadecylammonium bromide（DDA）, E1C0 + DDA, E1C0 + MPL, E1C0 + IFA, E1C0, physiological saline and MPL + DDA for 3 times, respectively, each at an interval of 2 weeks. The dosages of E1C0, MPL, DDA and IFA for immunization were 100 μg, 25μg, 250μg and 100 μl, respectively. The mice were killed 4 weeks after the last immunization, and their spleens were collected aseptically, from which splenic lymphocytes were isolated, cultured with E1C0, then determined for proliferation level by MTT method, and for IFNlevel in culture supernatant by ELISA. Three batches of BALB/c mice were immunized with the screened adjuvant combined with antigen, and determined for IFNγ induced. Results The linear range, sensitivity and CV value of developed SABC-ELISA system were 40 ~ 2 560 pg / ml（R > 0. 98）, 40 μg/ml and less than 15%respectively, by which all the detection results of IFN酌in rat, guinea pig and rabbit sera were negative. The stimulating indexox（SIs） of specific lymphocyte proliferation in E1C0 + MPL + DDA, E1C0 + IFA and E1C0 + DDA groups were significantly higher than those in physiological saline group （P < 0. 01）. The IFN酌level secreted by lymphocytes in E1C0 + MPL + DDA group after stimulation with E1C0 in vitro was significantly higher than those in other groups （P < 0. 001）. No significant differences were observed in IFNγ levels induced in 3 batches of mice in E1C0 + MPL + DDA group（P > 0. 05）. Conclusion The immunization with E1C0 in a combination with MPL and DDA elicited a strong Th1 cellular immune response in mice. Mouse model for evaluation of activity of E1C0 antigen was successfully established

二十辊轧机齿轮箱失效分析及改造实践

对某钢厂二十辊冷轧板卷取机齿轮箱设备失效进行分析,对轮齿进行了应力分析,并重新计算,进行齿轮国产化设计制造,较好地满足了企业正常生产

JUNO Sensitivity on Proton Decay p→νˉK+p\to \bar\nu K^+ Searches

The Jiangmen Underground Neutrino Observatory (JUNO) is a large liquid scintillator detector designed to explore many topics in fundamental physics. In this paper, the potential on searching for proton decay in $p\to \bar\nu K^+$ mode with JUNO is investigated.The kaon and its decay particles feature a clear three-fold coincidence signature that results in a high efficiency for identification. Moreover, the excellent energy resolution of JUNO permits to suppress the sizable background caused by other delayed signals. Based on these advantages, the detection efficiency for the proton decay via $p\to \bar\nu K^+$ is 36.9% with a background level of 0.2 events after 10 years of data taking. The estimated sensitivity based on 200 kton-years exposure is $9.6 \times 10^{33}$ years, competitive with the current best limits on the proton lifetime in this channel