摘要
目的探讨酶联免疫吸附试验中普遍存在的操作问题采用质控方法研究的可行性。方法 94份血清标本均为HBcAb阴性,采用两种不同加样顺序对竞争抑制法存在的操作问题进行对比实验,观察其差异;对酶联免疫检测实验操作的关键步骤进行质控,以CEA检测试剂为例,回顾性将北京军区总医院中心实验室15个不同批次的CEA标准血清结果总结分析,计算批间变异系数;选CEA临界值血清数份,选取范围在5.0～5.5ng/ml,检测45个批次的平均值、标准差和变异系数。结果用两种不同顺序加样方法检测HBcAb的结果显示,两种方法有很大差异,交替加样法未见假阳性结果,而顺序加样法有17.02%(16/94)出现阳性反应,属于假阳性结果。两种不同加样顺利做出的结果有统计学意义(P<0.01)。CEA标准血清分为0,2,5,15,50,75ng/ml,对15个不同批次试剂盒所做的标准曲线统计处理,其620nm吸光度变异系数分别为11.8%、11.8%、9.22%、8.78%、8.46%和4.23%。该变异反映出实验室常规条件下的变异,操作者常规操作水平。临界值血清经过45次不同批次实验,其变异系数只有3.68%,均值±标准差为0.3633±0.0134。结论对酶联免疫试验中采用竞争抑制法的项目应采用交替加样法以便减少假阳性;对ELISA实验操作中的几个关键步骤进行质控,能够保证实验的准确性。
Objective To approach the feasibility of studying the general operating problems in ELISA with the quality control method.Methods Ninty-four serum samples with HBcAb（-）were collected for the assay.The difference between the two different procedure of the competitive immunoassay was observed.With the CEA kit,we retrospectively assessed the quality control of the key points of the ELISA procedure.We summarized the results of 15 different production lots of the CEA standards serum and figured out the interassay CV.We chose the sera with CEA level ranged 5.0-5.5 ng/ml and calculated the mean value,SD and CV of the 45 lots of kits.Results It showed that the results were significantly different between the two methods on assaying the HBcAb（P0.01）.There was no false-positive by the alternative sample-adding,but 17.02%（16/94）false-positive by the orderly sample-adding.At the standard value of 0,2,5,15,50,75 ng/ml,the standards curve of CEA of the 15 different production lots were statistically analyzed.The CVs of the OD620 were 11.8%,11.8%,9.22%,8.78%,8.46% and 4.23% respectively.The CV value reflect the general skill of the operator under the routine condition of the lab.The cut-off value of the serum CEA was corrected by the test of 45 lots,and the CV was only 3.68%（0.3633±0.0134）.Conclusions The alternative sample-adding method in ELISA might reduce the rate of the false positive; Quality control of the key points of the procedure of ELISA could ensure the accuracy of the test.
出处
《中华临床医师杂志（电子版）》
CAS
2010年第10期95-98,共4页
Chinese Journal of Clinicians(Electronic Edition)
关键词
酶联免疫吸附测定
癌胚抗原
肝炎抗体
乙型
实验室技术和方法
Enzyme-linked immunosorbent assay
Carcinoembryonic antigen
Hepatitis B antibodies
Laboratory techniques and procedures
