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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χ~2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 Compared with the traditional manual washing method and horizontal injection liquid washing method, the application effect of automatic vertical washing method in enzyme-linked immunosorbent assay was evaluated. Methods The performance of the automatic vertical plate washer plate washer was evaluated, including the determination of residual liquid in the micropore, the possibility of blocking the needle and the possibility of cross-contamination. The results of quantitative detection of hepatitis B virus surface antigen chemiluminescence were negative, weak positive, positive, strong positive in each of the four groups of 30 samples, the use of enzyme-linked immunosorbent assay Hepatitis B virus surface antigen diagnostic kit detection, Including the steps of the washing plate respectively using manual washing method, horizontal liquid injection washing method, automatic vertical washing method three ways, the results read by the microplate reader, washing time, record data using χ ~ 2 test Statistical analysis. Results In this study, the average amount of residual liquid in micropore of ELISA plate was 0.0087 ± 0.0002μL, <0.01μL. The automatic vertical plate washer was used for 6 months (at least 2 times a week) , 96 needles did not appear blocking phenomenon, washing machine 6 months before and after the length of the water column were 6.53 ± 0.27cm, 6.58 ± 0.31cm, no significant difference between the two groups of data; positive control and negative control Cross-detection of products, and cross-detection of positive mixed serum and negative mixed serum cross-contamination did not occur. 120 cases of clinical samples using three kinds of washing the same qualitative test results, of which 30 cases of weak positive samples, one case of false negative and from the same sample; three wash plate method negative, weak positive, positive, strong positive samples The difference was not statistically significant (P> 0.05). Manual washing method, the level of liquid washing method, automatic washing method of the vertical washing time were 8min, 5min, 20s. Conclusion The automatic vertical plate washing method can meet the requirement of washing plate in the clinical testing of enzyme-linked immunosorbent assay.