酶联免疫吸附实验中全自动立式洗板法应用效果评价  

The Application Evaluation of Automatic Vertical Plate Washer Method in the Enzyme-linked Immunosorbent Assay

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

机构地区：[1]首都医科大学附属北京天坛医院,北京100050 [2]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 [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1068-1072,共5页Labeled Immunoassays and Clinical Medicine

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

摘　　要：目的 与传统手工洗板法、水平注液式洗板法相比较,评价酶联免疫吸附实验中全自动立式洗板法的应用效果.方法 评价全自动立式洗板法洗板机的性能,包括微孔中液体残留量测定、针头堵孔可能性检测、交叉污染可能性检测.收集乙型肝炎病毒表面抗原化学发光法定量检测结果分别为阴性、弱阳性、阳性、强阳性的四组样本各30例,使用酶联免疫吸附实验原理乙型肝炎病毒表面抗原诊断试剂盒检测,其中洗板步骤分别使用手工洗板法、水平注液式洗板法、全自动立式洗板法三种方法,由酶标仪读取结果,记录洗板时间,数据利用r检验进行统计学分析.结果 本研究中全自动立式洗板法酶标板微孔中液体残留量均值为0.0087±0.0002μL,＜0.01μL;全自动立式洗板机使用6个月（每周至少使用2次）,96个针头未出现堵孔现象,洗板机6个月前后的直线水柱长度分别为6.53±0.27cm、6.58 ±0.31cm,两组数据差异无统计学意义;阳性质控品和阴性质控品交叉检测,及阳性混合血清和阴性混合血清交叉检测未出现交叉污染现象.120例临床样本利用三种洗板法的定性检测结果一致,其中30例弱阳性样本,均有1例假阴性且来源于同一样本;三种洗板法检测阴性、弱阳性、阳性、强阳性样本的测定结果差异无统计学意义（P＞0.05）.手工洗板法、水平注液式洗板法、全自动立式洗板法的洗板时间分别为8min、5min、20s.结论 全自动立式洗板法可满足酶联免疫吸附实验临床检测中洗板要求.Objective To evaluate the application effect of automatic vertical plate washer method compared with traditional manual plate washer method and horizontal injection liquid type plate washer method in the enzyme-linked immunosorbent assay. Methods Evaluating the performance of the instrument which use the automatic vertical plate washer method, including the measurement of liquid residue in microporous, needle blocking probability detection and cross contamination possibility. A total of 120 serum samples with the quantitative test results of the hepatitis B surface antigen bychemiluminescence methodwere collected. The samples were divided into four groups equally, which were negative, weak positive, positive, strong positive.Each group had 30 cases. The hepatitis B virus surface antigen diagnostic kits based on enzyme-linked immunosorbent assayprinciple were used, the plates were washed by manual plate washer method, horizontal injection liquid type plate washer method, automatic vertical plate washer methodrespectively. The results were read by enzyme reader and the plate washing time was recorded. The data were analyzed by χ2 test. Results this study ,the average of liquid residue in microporous was 0. 0087 ± 0. 0002μ L, less than 0.01 μL. After using automatic vertical plate washer for six months （at least twice a week）, 96 needles haven＇ t been blocked. The linear water column length of the washer was 6.53 ± 0.27 cm, it was 6.58 ± 0.31 cm six months later. The data of two groups had no statistical difference. The cross test of positive quality product and negative quality product did not have cross contamination. So did the cross test of positive mixed samples and negative mixedsamples. The qualitative test results of 120 clinical samples by using three kinds of plate washer method were consistent. In the 30 cases of weakly positive samples, all three method could detect one false negative result respectivelyand the serum came from the same sample, the detection of negative, weak positive ,positiv

关 键 词：全自动立式洗板法 手工洗板法 水平注液式洗板法 酶联免疫吸附实验 乙型肝炎病毒表面抗原 

分 类 号：R446.6[医药卫生—诊断学]