应用快速孵育法优化酶联免疫吸附实验过程探讨——以乙型肝炎病毒表面抗原检测为例  

Evaluation of Rapid Incubation Optimization Process of Enzyme-linked Immunosorbent Assay on Hepatitis B Surface Antigen Detection

作　　者：刘丽[1] 何露[2] 孔凡虹 曹敬丽[1] 贾子琪[2] 刘文松[2] 许震[2] 王雅杰[1,3] 

机构地区：[1]首都医科大学附属北京天坛医院临床医学研究实验室,北京100050 [2]首都医科大学附属北京天坛医院,北京100050 [3]首都医科大学附属北京天坛医院检验科,北京100050

出　　处：《标记免疫分析与临床》2016年第9期1073-1076,共4页Labeled Immunoassays and Clinical Medicine

基　　金：家自然科学基金(81572474);首都医科大学校长基金(2015JYY80);北京市中医药科技发展资金项目(JJ2015-14)

摘　　要：目的利用快速孵育法优化酶联免疫吸附实验操作流程,通过缩短孵育时间,以提高工作效率。方法收集化学发光法定量检测乙肝表面抗原阳性高值样本30例、阳性中值样本30例、阳性临界值样本30例、阴性样本30例,使用ELISA试剂盒说明书推荐方法和快速孵育法同时进行检测,比较检测结果相关性。结果使用酶标板孵育器将孵育时间缩短至原来的1/2、1/3时,ELISA试剂盒说明书推荐方法、快速孵育法与化学发光法检测结果符合率100%;当孵育时间缩短至原来时间的1/4时,两种方法与化学发光法阴性、高值和中值样本检测结果符合率100%;临界值样本检测时,ELISA试剂盒说明书推荐方法与化学发光法检测结果符合率100%,快速孵育法与化学发光法结果 6例不一致,检测结果符合率80%。结论通过实验证实,酶联免疫吸附实验时,快速孵育法可满足临床检测,可将孵育时间缩短至原试剂盒推荐时间的1/3或1/2,可提高工作效率。Objective To evaluate the rapid incubation of enzyme- linked immunosorbent assay （ ELISA ） on hepatitis B surface antigen. The goal is to reduce the incubation time in order to improve operational efficiency. Methods Hepatitis B surface antigen positive samples detected by chemiluminescence were used in the study. The samples were divided into 3 groups with 30 cases in each group based on the posititivity value, highly positive, borderline positive, negative samples. All samples were tested with ELISA kit both by standard method recommended by manufacturer and by fast incubation method. The results were comparated and analyzed. Results When incubation time was shortened to one-half and one-third of standard time, the results from rapid incubation method and chemiluminescence detection matched 100%. When the time is shortened to one quarter of standard time the results from rapid incubation method and chemiluminescence detection matched 100% in all highly positive samples. For the borderline samples ,results from both methods matched 100%. For fast incubation method, there were 6 results which were inconsistent with chemiluminescence, 80%. Conclusion The study confirmed that ELISA rapid incubation method satisfied the use of clinical testing;the incubation time may be reduced to one-third or one-half of recommended time. The rapid incubation method will improve the work efficiency in clinical laboratories.

关 键 词：酶联免疫吸附实验 快速孵育法 

分 类 号：R446.6[医药卫生—诊断学] R512.62[医药卫生—临床医学]