[1]胡纪文,杨锡琴,宗克亮,等.人羧基肽酶H不同区段抗原的克隆表达及其初步应用[J].现代检验医学杂志,2015,30(04):10-13.[doi:10.3969/j.issn.1671-7414.2015.04.003]
 HU Ji-wen,YANG Xi-qin,ZONG Ke-liang,et al.Cloning,Expression and Diagnostic Application of Different Fragments of Human Carboxypeptidase[J].Journal of Modern Laboratory Medicine,2015,30(04):10-13.[doi:10.3969/j.issn.1671-7414.2015.04.003]
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人羧基肽酶H不同区段抗原的克隆表达及其初步应用()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第30卷
期数:
2015年04期
页码:
10-13
栏目:
论著
出版日期:
2015-08-10

文章信息/Info

Title:
Cloning,Expression and Diagnostic Application of Different Fragments of Human Carboxypeptidase
文章编号:
1671-7414(2015)04-010-04
作者:
胡纪文1杨锡琴2宗克亮3郭兰芹4宋晓国2王国华2刘喜明5朱翠侠2赵琰枫2 冯晓燕2张贺秋2
1.深圳市罗湖区中医院,广东深圳 518001;
2.军事医学科学院基础医学研究所,北京 100850;
3.风帆集团职工医院,河北保定 071000;
4.华北石油总医院,河北任丘 062552;
5.中国中医科学院广安门医院,北京 100053
Author(s):
HU Ji-wen1YANG Xi-qin2ZONG Ke-liang3GUOLan-qin4SONG Xiao-guo2 WANG Guo-hua2LIU Xi-ming5ZHU Cui-xia2ZHAO Yan-feng2FENG Xiao-yan2ZHANG He-qiu2
1.Chinese Medicine Hosptial of Luohu District,Guangdong Shenzhen 518001,China;
2.Beijing Institute of Basic Medical Sciences,Beijing 100850,China;
3.the Worker's Hospital of Sail Group,Hebei Baoding 071000,China;
4.the General Hospital o
关键词:
人羧基肽酶H 自身抗体 新诊断2型糖尿病 成人隐匿性自身免疫性糖尿病
分类号:
R587.1; Q786
DOI:
10.3969/j.issn.1671-7414.2015.04.003
文献标志码:
A
摘要:
目的 构建人羧基肽酶H不同区段抗原的原核表达载体,诱导表达获得重组蛋白,初步验证人羧基肽酶H不同抗原区段在羧基肽酶H自身抗体检测中的应用价值。方法 应用RT-PCR方法调取目的基因,构建相应的原核表达质粒,转化大肠埃希氏菌诱导表达重组蛋白,用重组蛋白作为包被抗原建立人羧基肽酶H自身抗体的间接ELISA方法,评价羧基肽酶H抗原在检测新诊断2型糖尿病患者抗羧基肽酶H自身抗体中的应用价值。结果 获得了羧基肽酶H三种不同区段抗原,其中42~476氨基酸区段为较理想的抗原。以该全长抗原作为包被抗原检测95例新诊断2型糖尿病患者,羧基肽酶H自身抗体阳性率为8.42%。结论 原核克隆表达的人羧基肽酶H 42~476氨基酸区段抗原具有良好的抗原性,可作为成人隐匿性自身免疫性糖尿病鉴别诊断的候选抗原。
Abstract:
Objective To obtain different fragments of human carboxypeptidase H,and evaluate the diagnostic application of the recombination carboxypeptidase H in detecting autoantibody.Methods The coding gene of carboxypeptidase H was obtained by RT-PCR.The corresponding prokaryotic expression vectors were constructed and transformed into E.coli to induce the expression of the recombination different fragments of carboxypeptidase H.Using these antigen fragments as the coating antigens,the enzyme-linked immunosorbent assay(ELISA)was established for the detection of carboxypeptidase H autoantibody in 95 newly diagnosed type 2 diabetes patients.Results Three fragments of human carboxypeptidase H were obtained,in which the 42~476aa fragment antigen was ideal one.Using the full-length carboxypeptidase H as coating antigen,the positive rate of carboxypeptidaseH autoantibody was 8.42%.Conclusion Because of the favorable antigenicity,the 42~476aa fragment antigen of carboxypeptidase H couldbe the candidate antigen for discrimination and diagnosis of latent autoimmune diabetes in adults.

参考文献/References:

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备注/Memo

备注/Memo:
基金项目:十二五国家高技术研究发展计划(863计划)课题,编号2011AA02A113; 深圳市伯劳特生物制品有限公司委托课题。
作者简介:胡纪文(1967-),男,医学学士,副主任技师,从事免疫学检验及检验方法学研究,Tel:13620208790,E-mail:shenzhenaw@163.com;
共同第一作者简介:杨锡琴(1985-),女,硕士,助理研究员,从事诊断试剂研制,Tel:15210343247,E-mail:yangxiqin001@163.com。
共同第一作者简介:宗克亮(1974-),男,硕士,副主任医师,从事内分泌系统疾病研究,Tel:13930242582,E-mail:zklbd@163.com。
通讯作者:冯晓燕(1973-),女,博士,副研究员,主要研究方向为疾病诊断标志物筛选与诊断试剂研制,Tel:13011821686,E-mail:xyfeng2002@126.com。
更新日期/Last Update: 2015-08-10