酶联免疫吸附实验中全自动立式洗板法应用效果评价
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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 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 methodw ere 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 χ~2test.Results this study,the average of liquid residue in microporous was 0.0087±0.0002μL,less than 0.01μL.After using automatic vertical plate washer for six months(at least tw ice 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,positive,strong positive samples had no statistical difference by using three plate washer method(P>0.05).The time of washing a plate by using three kinds of plate washer method was 8min,5min,20 s respectively.Conclusion The automatic vertical plate washer method can meet the clinical requirement of washing plates in the enzyme-linked immunosorbent assay.
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 methodw ere 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 χ~2test.Results this study,the average of liquid residue in microporous was 0.0087±0.0002μL,less than 0.01μL.After using automatic vertical plate washer for six months(at least tw ice 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,positive,strong positive samples had no statistical difference by using three plate washer method(P>0.05).The time of washing a plate by using three kinds of plate washer method was 8min,5min,20 s respectively.Conclusion The automatic vertical plate washer method can meet the clinical requirement of washing plates in the enzyme-linked immunosorbent assay.
引文
[1]Abdelwahab M,Loa C C,Wu C C,et al.Recombinant nucleocapsid protein-based enzyme-linked immunosorbent assay for detection of antibody to turkey coronavirus.J Virol Methods,2015,217:36-41.
[2]Thiha A,Ibrahim F.A Colorimetric enzyme-linked immunosorbent assay(ELISA)detection platform for a point-of-care dengue detection system on a Lab-on-Compact-Disc.Sensors(Basel),2015,15(5):11431-11441.
[3]顾友祥.酶联免疫吸附试验检测乙型肝炎病毒表面抗原的全面质量控制前后对比分析.检验医学与临床,2010,7(20):2194-2195.
[4]王静.ELISA快速法检测HBs Ag时手工洗板和洗板机洗板的结果比较.中国医药导报,2010,7(9):69,73.
[5]王红.酶联免疫吸附试验检测丙型肝炎病毒抗体在酶标仪应用中的探讨.检验医学与临床,2012,9(10):1234-1235.
[6]武利庆,杨彬,丁峰元,等.基于吸光度测定的洗板机校准方法及其不确定度评定.计量技术,2012,(5):46-48.
[7]Le T,Yu H,Niu X.Detecting quinoxaline-2-carboxylic acid in animal tissues by using sensitive rapid enzyme-linked immunosorbent assay and time-resolved fluoroimmunoassay.Food Chem,2015,175:85-91.
[8]张桂芳,张永峰,张向峰,等.EDTA-K2对双抗原夹心ELISA检测HIV抗体的影响.检验医学,2012,27(8):676-678.
[2]Thiha A,Ibrahim F.A Colorimetric enzyme-linked immunosorbent assay(ELISA)detection platform for a point-of-care dengue detection system on a Lab-on-Compact-Disc.Sensors(Basel),2015,15(5):11431-11441.
[3]顾友祥.酶联免疫吸附试验检测乙型肝炎病毒表面抗原的全面质量控制前后对比分析.检验医学与临床,2010,7(20):2194-2195.
[4]王静.ELISA快速法检测HBs Ag时手工洗板和洗板机洗板的结果比较.中国医药导报,2010,7(9):69,73.
[5]王红.酶联免疫吸附试验检测丙型肝炎病毒抗体在酶标仪应用中的探讨.检验医学与临床,2012,9(10):1234-1235.
[6]武利庆,杨彬,丁峰元,等.基于吸光度测定的洗板机校准方法及其不确定度评定.计量技术,2012,(5):46-48.
[7]Le T,Yu H,Niu X.Detecting quinoxaline-2-carboxylic acid in animal tissues by using sensitive rapid enzyme-linked immunosorbent assay and time-resolved fluoroimmunoassay.Food Chem,2015,175:85-91.
[8]张桂芳,张永峰,张向峰,等.EDTA-K2对双抗原夹心ELISA检测HIV抗体的影响.检验医学,2012,27(8):676-678.