电化学发光免疫分析法检测人胰岛素原方法的建立  

Establishment of electrochemiluminescence immunoassay to detect proinsulin levels in human serum

作　　者：张玫[1] 施绍瑞[1] 张林[1] 熊中云[1] 黄明洁[1] 安振梅[1] 

机构地区：[1]四川大学华西医院实验医学科激素检测室,成都610041

出　　处：《中华检验医学杂志》2009年第9期1015-1018,共4页Chinese Journal of Laboratory Medicine

摘　　要：目的建立一种快速、特异、灵敏的检测人胰岛素原（PI）电化学发光免疫分析法（ECLIA）。方法用链霉亲和素包被的微粒、生物素化的抗胰岛素单克隆抗体、钌标记的抗C肽单克隆抗体组成PIECLIA试剂盒，并对其准确性、灵敏度、特异性等效能进行方法学评价，同时用所建方法与放射免疫分析法（mA）检测和对比分析44例2型糖尿病患者和60名健康志愿者PT水平。结果用自建ECLIA检测PI的平均回收率为103．8％。检测低限为0．38pmol／L，生物检测限在0．25—0．5pmol／L之间，功能灵敏度为0．5pmol／L，与500μU／ml胰岛素和450ng／ml C肽无交叉反应，总不精密度小于5．0％，分析测量范围为2．22—169．68pmol／L；与IliA比较，相关性好（r=0．967，P=0．000），自建ECLIA检测Pl的95％参考区间为1．07—15．54pmol／L，ECLIA检测人PI时间（18min）比RIA方法短（72h）。临床检测结果显示，2型糖尿病患者血清空腹PI[6．72（3．61～11．92）pmo]／L]高于健康对照组[5．72（2．65～8．74）pmol／L，Z=-2．023，P〈0．05]。结论自建ECLIA检测PI法特异性高，灵敏度好，检测速度快，无放射性污染，在诊断2型糖尿病中有较好的应用价值。Objective To establish a electrochemiluminescence immunoassay （ECLIA） for detecting proinsulin（PI） levels in human serum. Methods PI ECLIA kit contains biotinylated monoclonal insulin-specificantibody, ruthenium labeled monoclonal C-peptide specific antibody, and streptavidin-coated microparticles. Its method performance, such as sensitivity, specific, accuracy, was evaluated. Then, the levels of PI in clinical samples were detected by ECLIA, including 44 type 2 diabetes and 60 healthy volunteers. Results Average recovery of ECLIA was 103.8% with a lower detection limit of 0. 38 pmol/L and biologic detection limit between 0. 25 pmol/L and 0. 5 pmol/L, and the function sensitivity was 0. 5 pmol/L. The human insulin and human C-peptide did not cross-react at 500 μU/ml recovery of ECLIA was 103. 8% with a lower detection limit of 0. 38 pmol/L and biologic detection limit between 0. 25 pmol/L and 0. 5 pmol/L, and the function sensitivity was 0. 5 pmol/L. The human insulin and human C-peptide did not cross-react at 500 i^U/ml and 450 ng/ml respectively. The total coefficient of variation of the ECLIA were less than 5.0% , and the analytical measure range （AMR） was between 2. 22 and 169. 68 pmol/L. There was a good correlation （ r = 0. 967, P = 0. 000 ） between ECLIA and radio immunoassay （RIA）. The 95% reference value for human proinsulin was 1.07-15.54 pmol/L. The plasma levels of PI in type 2 diabetes were significantly higher than those in control healthy group[6. 72（3.61 - 11.92） pmol/L vs 5.72（2. 65 - 8.74） pmol/L, Z = - 2. 023, P 〈 0. 05 ]. Conclusions The monoclonal- based ECLIA is a sensitive, specific, and rapid method and no radiocontamination. It can be used to detect human serum proinsulin in type 2 diabetes.

关 键 词：胰岛素原 电化学 发光 免疫测定 

分 类 号：R686[医药卫生—骨科学]