抗蛋白酶3抗体检测及影响因素的研究  

Research for influence factors on detection of antibodies to proteinase-3

作　　者：胡学芳[1] 李小峰[1] 朱爱萍[2] 郭红霞[3] 雷智[4] 

机构地区：[1]山西医科大学第二医院风湿免疫科,030001 [2]山西医科大学第二医院检验科,030001 [3]太原市红十字血液中心血库 [4]太原市红十字血液中心成分制备科

出　　处：《中国药物与临床》2004年第10期752-754,共3页Chinese Remedies & Clinics

基　　金：山西省高校科技研究开发基金资助项目(2001-10)

摘　　要：目的研究血清样本稀释度、封闭液、孵育条件、洗涤条件及酶标板包被后保存时间对血清抗蛋白酶3(PR3)抗体检测结果的影响。方法采用酶联免疫吸附试验(ELISA)对159份血清进行抗PR3抗体检测,并用受试者工作特征曲线(ROC曲线)分析确定阳性界限(cut-off)值。结果血清样本最佳稀释度为1∶50;5%脱脂奶粉封闭酶标板能有效地降低本底值;孵育缓冲液中加入牛血清白蛋白及脱脂奶粉比单用磷酸盐缓冲液(PBS)对测定值有明显影响。经ROC曲线分析实验的阳性临界值为正常阴性对照吸光度(A)值的2.5倍,灵敏度和特异性分别为84.62%和98.46%。结论血清抗PR3检测结果受血清样品稀释、封闭液、孵育条件及阳性临界值的选择等多因素的影响。Objective To investigate the influence factors on the detective level of serum anti-proteinase 3 (anti-PR3) antibody by analyzing the absorptance at various conditions, including sample dilutions, blocking materials, incubation condition and interval time between blocking and detection. Methods Anti-PR3 from 159 serum samples (13 patients and 146 controls) was assayed by ELISA, and the cut-off values were defined by ROC curve obtained. Results Serum dilution at 1∶50 was suitable for the detection; absorptance of background blocked by 5% defatted milk powder was decreaced significantly; the incubation buffer containing bovine serum albumin or 5% defatted milk powder influenced the detection more obviously than that containing PBS only; the area under curve (AUC) was 0.928 (95% CI 0.834~1.021); cut-off value of absorptance was 0.235; sensitivity and specificity were 84.62% and 98.46% respectively. Conclusion The detection of anti-PR3 can be influenced by serum dilution, blocking materials, incubation condition and the choice of cut-off point.

关 键 词：血清 抗蛋白酶 阳性 PR 抗体检测 孵育 酶联免疫吸附试验(ELISA) 封闭液 包被 稀释度 

分 类 号：R446.6[医药卫生—诊断学]