中华糖尿病杂志
中華糖尿病雜誌
중화당뇨병잡지
CHINES JOURNAL OF DLABETES MELLITUS
2010年
1期
24-28
,共5页
宋晓国%张贺秋%王国华%楚晓燕%刘喜明%陈坤%朱翠侠%戴振华%方平%冯晓燕
宋曉國%張賀鞦%王國華%楚曉燕%劉喜明%陳坤%硃翠俠%戴振華%方平%馮曉燕
송효국%장하추%왕국화%초효연%류희명%진곤%주취협%대진화%방평%풍효연
1型糖尿病%重组人谷氨酸脱羧酶65%谷氨酸脱羧酶自身抗体%酶联免疫吸附测定法
1型糖尿病%重組人穀氨痠脫羧酶65%穀氨痠脫羧酶自身抗體%酶聯免疫吸附測定法
1형당뇨병%중조인곡안산탈최매65%곡안산탈최매자신항체%매련면역흡부측정법
Type 1 diabetes mellitus%Recombination human glutamic acid decarboxylase 65%Glutamic acid decarboxylase autoantibody%Enzyme-linked immunosorbent assay
目的 构建人重组谷氨酸脱羧酶65(GAD65)基因不同区段原核表达载体,诱导表达获得重组蛋白,并初步验证GAD65不同抗原区段在1型糖尿病GAD自身抗体检测中的价值.方法 应用巢式逆转录聚合酶链式反应(RT-PCR)技术调取目的 基因,构建相应的原核表达质粒,转化大肠杆菌E.coli HB101,诱导表达获得纯化重组蛋白,用重组蛋白作为包被抗原,初步建立检测GAD自身抗体的酶联免疫吸附测定法(ELISA)方法,评价各片段在1型糖尿病诊断中的价值.结果 获得了4种可被1型糖尿病患者血清识别的重组人GAD抗原区段,其中GAD65(180-585)抗原区段具有很好的特异性,检出率为55.3%,是首选的抗原区段.结论 所选重组人GAD65(180-585)抗原区段具有良好的抗原性,可作为1型糖尿病患者辅助诊断试剂的候选抗原.
目的 構建人重組穀氨痠脫羧酶65(GAD65)基因不同區段原覈錶達載體,誘導錶達穫得重組蛋白,併初步驗證GAD65不同抗原區段在1型糖尿病GAD自身抗體檢測中的價值.方法 應用巢式逆轉錄聚閤酶鏈式反應(RT-PCR)技術調取目的 基因,構建相應的原覈錶達質粒,轉化大腸桿菌E.coli HB101,誘導錶達穫得純化重組蛋白,用重組蛋白作為包被抗原,初步建立檢測GAD自身抗體的酶聯免疫吸附測定法(ELISA)方法,評價各片段在1型糖尿病診斷中的價值.結果 穫得瞭4種可被1型糖尿病患者血清識彆的重組人GAD抗原區段,其中GAD65(180-585)抗原區段具有很好的特異性,檢齣率為55.3%,是首選的抗原區段.結論 所選重組人GAD65(180-585)抗原區段具有良好的抗原性,可作為1型糖尿病患者輔助診斷試劑的候選抗原.
목적 구건인중조곡안산탈최매65(GAD65)기인불동구단원핵표체재체,유도표체획득중조단백,병초보험증GAD65불동항원구단재1형당뇨병GAD자신항체검측중적개치.방법 응용소식역전록취합매련식반응(RT-PCR)기술조취목적 기인,구건상응적원핵표체질립,전화대장간균E.coli HB101,유도표체획득순화중조단백,용중조단백작위포피항원,초보건립검측GAD자신항체적매련면역흡부측정법(ELISA)방법,평개각편단재1형당뇨병진단중적개치.결과 획득료4충가피1형당뇨병환자혈청식별적중조인GAD항원구단,기중GAD65(180-585)항원구단구유흔호적특이성,검출솔위55.3%,시수선적항원구단.결론 소선중조인GAD65(180-585)항원구단구유량호적항원성,가작위1형당뇨병환자보조진단시제적후선항원.
Objective To obtain different fragments of human glutamic acid decarboxylase 65 and to evaluate the diagnostic application for type 1 diabetes mellitus of the recombination human glutamic acid decarboxylase 65. Methods The coding gene of the glutamic acid decarboxylase 65 was obtained by RT-PCR. The corresponding prokaryotic expression vectors pBVILl/GAD65 were constructed and transformed into E.coli HBI01 to inducing the expression of the recombination human glutamic acid decarboxylase 65. Using these antigen fragments as the coating antigens, the enzyme-linked immunosorbent assay ( ELISA) was established for the detection of the glutamic acid decarboxylase autoantibody. Results The obtained four fragments of human glutamic acid decarboxylase 65 could react with the serum of type 1 diabetes mellitus patients. Because the specificity of the fragment of GAD65( 180-585) was the highest and the detection rate of this fragment was 55. 3% , the fragment of GAD65( 180-585) became the preferred antigen. Conclusion Because of the favourable antigenicity of the selected recombination human GAD65 (180-585), the GAD65 (180-585) could be the candidate antigen for developing the diagnostic reagent for type 1 diabetes mellitus.
