胃蛋白酶原双标记时间分辨荧光免疫分析及其初步临床应用  

Dual-label Time-resolved Fluoroimmunoassay for Pepsinogen Ⅰ and Pepsinogen Ⅱ and Its Preliminary Application

作　　者：张珏[1] 黄飚[1] 朱岚[1] 张艺[1] 刘海燕[1] 马智鸿[1] 郭力宁[1] 

机构地区：[1]江苏省原子医学研究所,无锡214063

出　　处：《生物化学与生物物理进展》2008年第4期471-476,共6页Progress In Biochemistry and Biophysics

基　　金：江苏省科学技术厅社会发展资助项目(BS2006015)~~

摘　　要：采用钐(Sm3+)标记抗胃蛋白酶原Ⅰ单克隆抗体(PGⅠ)及铕(Eu3+)标记抗胃蛋白酶原Ⅱ单克隆抗体(PGⅡ),建立了双标记时间分辨荧光免疫分析法(TRFIA),同时检测人血清PGⅠ和PGⅡ.将抗PGⅠ单克隆抗体8003#、抗PGⅡ单克隆抗体8101#以一定比例共包被于96孔微孔板上,用Sm3+标记抗PGⅠ单抗8016#、Eu3+标记抗PGⅡ单抗8102#,采用双抗体夹心法建立PGⅠ/PGⅡ的双标记TRFIA.标准曲线由TRFIA检测仪自带的Log-LogB函数处理.PGⅠ可测范围为0.2～300μg/L,批内和批间变异系数(CV%)分别为5.2%和8.1%,平均回收率为96.9%;PGⅡ可测范围为0.05～55μg/L,批内和批间变异系数(CV%)分别为7.1%和11.7%,平均回收率为103.7%;双标记PGⅠ/PGⅡ-TRFIA与PGⅠ-ELISA法、PGⅡ-ELISA法的测定结果高度相关,具有较好的一致性,相关系数分别为0.9426和0.9396.检测300例健康人员的PGⅠ为(157.3±51.0)μg/L,PGⅡ为(10.6±5.9)μg/L,PGⅠ/PGⅡ比值为(14.8±4.3)μg/L.PGⅠ的正常参考值范围在55.3～259.3μg/L之间,PGⅡ正常参考值为<23μg/L,PGⅠ/PGⅡ>6.双标记PGⅠ/PGⅡ-TRFIA方法的研究在国内外尚未见报道,是一种灵敏、简便、快速、经济并可用于大批量样品筛查的方法,有利于各种胃病的大规模普查筛选及患者病程监测.A dual-label time-resolved fluoroimmunoassay was established for simultaneously detecting pepsinogen Ⅰ （PGⅠ） and pepsinogen Ⅱ （PG Ⅱ） in human serum. Two capture monoclonal antibodies, 8003# of PGⅠ and 8101# of PGⅡ, were co-coated in 96 microtitration wells. The counterpart tracer monoclonal antibodies, 8016# of PGⅠ and 8102# of PGⅡ, were labeled with Eu3＋ and sm3＋-chelates, respectively. The samples were assayed by one-step sandwich protocol with the time-resolved fluorometry. The measurement ranges of PGⅠ were 0.2～300.0 μg/L with the within-run and between-run precision was 5.2% and 8.1%, and that of PGⅡ were 0.05～55.0 μg/L with the within-run and between-run precision was 7.1% and 11.7%, respectively. The average recovery rates of PGⅠ and PGⅡ were 96.9% and 103.7%, respectively. The results obtained by the dual-label assay agreed well with those by enzyme-linked immunosorbent assays of PGⅠ and PGⅡ, whose correlation ratio were 0.9426 of PGⅠ and 0.9396 of PGⅡ, respectively. The means of 300 healthy volunteers were （157.3 ± 51.0） μg/L for serum PGⅠ,（10.6 ± 5.9） μg/L for serum PGⅡ, and （14.8 ± 4.3） for the PGⅠ/PGⅡ ratio. The normal ranges of serum PGⅠ levels for healthy volunteers were 55.3～259.3 μg/L, those of serum PGⅡ levels were less than 23 μg/L, the PGⅠ/PGⅡ ratio was more than 6. The proposed dual-label TRFIA for simultaneous detection of PGⅠ and PGⅡ is a simple, sensitive, and rapid method. It could provide serology high-screening of the samples for gastric diseases and would allow investigations into the possible diagnostic value of analysis in various clinical condition.

关 键 词：胃蛋白酶原Ⅰ 胃蛋白酶原Ⅱ 双标记 时间分辨荧光免疫分析法 

分 类 号：R446.6[医药卫生—诊断学] R392.3[医药卫生—临床医学]