摘要
目的制备人催乳素(hPRL)化学发光免疫分析(CLIA)试剂盒,并进行验证。方法采用2株不同结合位点的抗hPRL单克隆抗体,1株用于包被微孔板,另1株用于标记HRP,建立双抗体夹心检测系统,配合化学发光底物组装成试剂盒,并进行各项技术指标的验证。结果试剂盒最佳定量范围在5～100ng/ml,标准曲线的相关系数r=0.9999,灵敏度为0.49ng/ml,平均回收率为100.9%,试验内和试验间变异系数分别小于6.3%和10.9%。试剂盒有效期可达1年,特异性良好,出现Hook效应的样品浓度为1500ng/ml。与进口化学发光试剂盒和放射免疫分析试剂盒对比检测84份临床标本的结果呈高度相关,相关系数分别为0.9537和0.9486。结论已成功制备灵敏、特异的hPRLCLIA试剂盒,可用于人催乳素的临床检测。
Objective To prepare and verify the chemiluminescent immunoassay（CLIA）kit for human proclactin（hPRL）. Methods A double antibody sandwich ELISA system was developed by using two kinds of anti-hPRL McAbs with different binding sites,of which one was used for coating of microtiter plate,and another was labeled with horseradish peroxidase（HRP）. A CLIA kit was prepared by using the developed ELISA system and chemiluminescent substrate and verified for various technical indexes. Results The optimal quantitative detection range,correlation coefficient of standard curve,sensitivity and mean recovery of the prepared kit were 5 ～ 100 ng / ml,0. 999 9,0. 49 ng / ml and 100. 9%,and the inter-and intra-variation coefficients of detection result were less than 6.3% and less than 10.9%,respectively. The prepared kit showed good specificity,and its validity period was 12 months. The antigen concentration resulting Hook effect was 1 500 ng / ml. The relation coefficients of detection result by the prepared kit to those by imported CLIA kit and by radioimmunoassay kit were 0. 953 7 and 0. 948 6 respectively. Conclusion A sensitive and specific CLIA kit for hPRL was successfully prepared,which might be used for clinical detection.
出处
《中国生物制品学杂志》
CAS
CSCD
2009年第4期382-384,共3页
Chinese Journal of Biologicals
关键词
人催乳素
化学发光免疫分析
试剂盒
验证
Human proclactin（hPRL）
Chemiluminescent immunoassay（CLIA）
Kit
Verification
