幽门螺杆菌黏附素A有效T和B联合抗原表位噬菌体展示和抗原性测定

目的 分析并确定幽门螺杆菌黏附素A(HpaA)分子中有效T细胞和B细胞联合(T/B)抗原表位.方法 重组表达HpaA(rHpaA)并免疫家兔制备抗血清.采用生物信息学技术预测和分析HpaA分子中T细胞和B细胞表位,PCB扩增T/B联合表位肽片段并构建其噬菌体展示系统.采用PEG/NaCl沉淀法提纯展示了T/B联合表位的重组噬菌体PⅢ蛋白(rPⅢ).分别以商品化幽门螺杆菌全菌IgG抗体和rHpaA抗血清为一抗,采用Western blot和ELISA对rPⅢ蛋白中展示的T/B联合表位进行筛选和鉴定.采用MTT检测rHpaA免疫小鼠脾细胞在不同重组噬菌体蛋白刺激下的增殖情况.结果 HpaA分子中共有5个T/B联合表位:HpaA10、HpaA37、HpaA79、HpaA116和HpaA143.所有T/B联合表位均成功地展示于M13噬菌体PⅢ蛋白表面.Western blot、ELISA和淋巴细胞增殖试验结果 均显示,HpaA116是优势抗原表位,HpaA37和HpaA79为有效抗原表位,HpaA10和HpaA143为无效抗原表位.结论 本研究成功地构建了幽门螺杆菌HpaA的T/B联合表位肽噬菌体展示系统.HpaA37和HpaA79,尤其是HpaA116是HpaA有效T/B联合抗原表位.     

微生物实验课教学方法改革的探索

实验技能目标的达标是目标教学体系的重要组成部分 ,也是培养合格的应用型实用人才的关键所在。我校在微生物“显微镜油镜的使用和保护”实验教学中 ,尝试了实验技能目标的层层达标教学方法 ,取得了较好的教学效果 ,现报告如下。1　教学对象和方法以我校 98级护理专业甲、乙两个平行班为对象 ,甲、乙班的人数分别为 51人和 53人。甲班采用改革法教学 ,乙班则采用传统法教学。改革的油镜教学法操作如下 :①告知教学目标 ,进行课前复习。该实验课的目标是学会显微镜油镜的使用和保护。课前首先向学生展开目标 ,要求学生有目标地进行原有相关知…     

肺结核病实验室快速诊断进展

结核病为严重危害人类健康的慢性传染病 ,80年代中后期出现世界范围内回升 ,特别面临的新情况是艾滋病毒 (ＨＩＶ)与结核分枝杆菌的双重感染者增多[1] 。为了控制其蔓延 ,实施有效的防治 ,进行早期诊断十分必要。传统的结核杆菌检测一般依赖于细菌学指标 ,其中痰涂片抗酸染色法     

幽门螺杆菌黏附素A有效T和B联合抗原表位噬菌体展示和抗原性测定

Objective To analyze and determine the efficient T- and B-combined (T/B) antigenic epitopes in Helicobacter pylori adhesin A. Methods Recombinant HpaA (rHpaA) was expressed for immunizing rabbit to generate antiserum. T- and B-cell epitopes in HpaA molecule were predicted by using bioinformatic technique. The segments to encode T/B combined epitope peptides were amplified by PCR and the phage display systems of T/B combined epitopes were subsequently constructed. PEG/NaCl precipitation method was applied to extract the recombinant phage PⅢ (rPⅢ) that displayed T/B combined epitopes. By using either commercial IgG against whole-cell of Helicobacter pylori or rHpaA antiserum as the primary antibody, the T/B combined epitopes displayed in rP Ⅲ s were screened and identified by Western blot and ELISA. MTT was applied to determine the proliferation of rHpaA-immunized mouse splenocytes after stimulation of the different recombinant rPⅢ proteins. Results In the HpaA molecule there were five T/B combined epitopes: HpaA10, HpaA37, HpaA79, HpaA116 and HpaA143. All the T/B combined epitopes were successfully displayed on the surface of PⅢ protein of phage M13. The results of Western blot, ELISA and MTT confirmed that HpaA116 was the predominant antigenic epitope, both HpaA37 and HpaA79 were the efficient antigenic epitopes. However, HpaA10 and HpaA143 were identified as ineffective antigenic epitopes. Conclusion The phage display systems of T/B combined epitope peptides of H. pylori adhesin A have been successfully generated in this study. HpaA37 and HpaA79, especially HpaA116 are the efficient T/B combined antigenic epitopes in HpaA of H. pylori.     

肺结核病实验室快速诊断进展

结核病为严重危害人类健康的慢性传染病 80年代中后期出现世界范围内回升特别面临的新情况是艾滋病毒(HIV)与结核分枝杆菌的双重感染者增多[1].为了控制其蔓延实施有效的防治进行早期诊断十分必要.传统的结核杆菌检测一般依赖于细菌学指标其中痰涂片抗酸染色法快速但灵敏度低而今仍被视为"金标准"的罗-琴(L-J)固体培养法虽然灵敏度高于染色法但培养时间至少要8周不能满足临床快速诊断的需要.本文就近年来有关肺结核实验室快速诊断的某些进展作一综述.     

幽门螺杆菌黏附素A有效T和B联合抗原表位噬菌体展示和抗原性测定

Objective To analyze and determine the efficient T- and B-combined (T/B) antigenic epitopes in Helicobacter pylori adhesin A. Methods Recombinant HpaA (rHpaA) was expressed for immunizing rabbit to generate antiserum. T- and B-cell epitopes in HpaA molecule were predicted by using bioinformatic technique. The segments to encode T/B combined epitope peptides were amplified by PCR and the phage display systems of T/B combined epitopes were subsequently constructed. PEG/NaCl precipitation method was applied to extract the recombinant phage PⅢ (rPⅢ) that displayed T/B combined epitopes. By using either commercial IgG against whole-cell of Helicobacter pylori or rHpaA antiserum as the primary antibody, the T/B combined epitopes displayed in rP Ⅲ s were screened and identified by Western blot and ELISA. MTT was applied to determine the proliferation of rHpaA-immunized mouse splenocytes after stimulation of the different recombinant rPⅢ proteins. Results In the HpaA molecule there were five T/B combined epitopes: HpaA10, HpaA37, HpaA79, HpaA116 and HpaA143. All the T/B combined epitopes were successfully displayed on the surface of PⅢ protein of phage M13. The results of Western blot, ELISA and MTT confirmed that HpaA116 was the predominant antigenic epitope, both HpaA37 and HpaA79 were the efficient antigenic epitopes. However, HpaA10 and HpaA143 were identified as ineffective antigenic epitopes. Conclusion The phage display systems of T/B combined epitope peptides of H. pylori adhesin A have been successfully generated in this study. HpaA37 and HpaA79, especially HpaA116 are the efficient T/B combined antigenic epitopes in HpaA of H. pylori.     

肺结核病实验室快速诊断进展

结核病为严重危害人类健康的慢性传染病,80年代中后期出现世界范围内回升,特别面临的新情况是艾滋病毒(HIV)与结核分枝杆菌的双重感染者增多[1].为了控制其蔓延,实施有效的防治,进行早期诊断十分必要.传统的结核杆菌检测一般依赖于细菌学指标,其中痰涂片抗酸染色法快速但灵敏度低,而今仍被视为"金标准"的罗-琴(L-J)固体培养法虽然灵敏度高于染色法,但培养时间至少要8周,不能满足临床快速诊断的需要.本文就近年来有关肺结核实验室快速诊断的某些进展作一综述.     

结核分枝杆菌特异性38kD多肽抗原提纯及其血清学诊断价值的研究

目的提纯结核分枝杆菌特异38kD多肽抗原,建立基于38kD多肽抗原(P38)的斑点金免疫渗滤法(DIGFA)检测结核病人血清标本,评价38kD多肽用于结核病血清学诊断抗原的实际价值。方法采用超声破碎、SDS-PAGE、切胶浸泡回收法提取结核分枝杆菌H37Rv株的38kD多肽抗原。建立以38kD多肽为包被抗原的DIGFA(P38-DIGFA),并检测147例临床确诊的活动性肺结核病人血清标本。同时建立以结核分枝杆菌脂阿拉伯甘露糖(LAM)为包被抗原的DIGFA(LAM-DIGFA)作为对照。结果结核分枝杆菌H37Rv株38kD多肽抗原提取物经SDS-PAGE后仅显示为单一的条带,并可与临床确诊的肺结核病人阳性血清标本呈阳性免疫印迹反应。P38-DIGFA和LAM-DIGFA检测肺结核病人血清标本的总阳性率分别为84.4%(124/147)和76.2%(112/147),其中59份痰涂片及培养均阳性的肺结核病人血清标本P38-DIGFA和LAM-DIGFA阳性率分别为88.1%(52/59)和84.7%(50/59),88份痰涂片及培养均阴性的肺结核病人血清标本P38-DIGFA和LAM-DIGFA阳性率分别为81.8%(72/88)和70.5%(62/88),60例气管炎病人血清标本阳性率分别为13.3%(8/60)和6.7%(4/60),120例健康人血清标本阳性率分别为5.8%(7/120)和5.0%(6/120)。各P38-DIGFA和LAM-DIGFA检测阳性率之间均无显著性差异。结论结核分枝杆菌38kD多肽作为抗原用于结核病血清学检测时,其敏感性和特异性与LAM相似,可应用于研发结核病血清学诊断试剂盒。     

“连贯性模块式”医学免疫学实验教学在临床医学生中的应用

文章通过建立＂连贯性模块式＂医学免疫学实验教学体系,初步探索出一套行之有效的免疫学实验教学模式,建立适合医学本科生进行创新思维和综合训练的实验教学平台,完善了实验教学质量保证体系。     

大学生面部蠕形螨感染情况调查及流行因素分析

目的 调查在校大学生面部蠕形螨感染状况,分析蠕形螨感染与相关流行因素的关系.方法 采用透明胶带粘贴法对200名大学生进行蠕形螨检测,同时通过问卷调查分析造成蠕形螨流行的相关因素与感染率的关系.结论 200名大学生蠕形螨的总感染率为38.5%,女生感染率为40.8%,男生感染率为34.7%,男女生感染率无显著性差异(x2=0.74,P＞0.05);77名感染者中感染度Ⅰ级(轻度)感染率为77.9%,Ⅱ级(中度)感染率为14.3%,Ⅲ级(重度)感染率为2.6%,Ⅳ级(极重度)感染率为5.2%;从面部肤质比较,油性皮肤感染率为43.9%,混合性皮肤为42.5%,干性皮肤为22.2%,油性和混合性皮肤感染率两者无显著性差异(x2=0.03,P＞0.05),而油性皮肤和干性皮肤两者感染率有显著性差异(x2=5.91,P＜0.05);面部皮肤健康者感染率为23.2%,不健康者(毛囊炎、酒渣鼻、痤疮等)为44.4%,两者比较差异有显著性(X2=7.68,P＜0.05).结论 大学生蠕形螨感染相对普遍,但以轻度感染为主,其阳性率高低与面部皮肤性质及面部皮肤有无疾患关系密切.