延长不同实验步骤温育时间对HBsAg酶免结果的影响

The Influences of extending incubate time of different experimental steps on the results of HBs Ag enzyme-linked immunosorbent assay

目的探讨延长不同实验步骤温育时间对乙肝表面抗原(hepatitis B surface antigen,HBs Ag)酶联免疫吸附实验(enzyme-linked immunosorbent assay,ELISA)结果的影响,从而保证检测结果的准确性和精确性。方法在其他实验参数不变的条件下,通过延长加酶前、加酶后、显色等步骤的温育时间,分别对强阳性、中阳性、弱阳性、阴性标本进行检测,所得数据采用SPSS 17.0软件进行相关分析。结果 1延长加酶前温育时间对强阳性和弱阳性标本的影响差异具有统计学意义,强阳性标本吸光度明显下降,弱阳性标本明显加强,并与延长的时间密切相关。2延长加酶后温育时间对各标本吸光度(阴性除外)影响最显著。3延长显色时间对实验结果影响并不明显。4延长各反应步骤温育时间对HBs Ag阴性标本吸光度影响均无统计学意义。结论选择正确的步骤适当延长温育时间可提高ELISA实验结果的准确性和弱阳性标本的检出率。

Objective To investigate the influences of extending incubate time of different experimental steps on the results of HBsAg enzyme -linked immunosorbent assay.Methods Without changing the condition of other experimental parameters,the incubate time before and after dispending enzyme and coloration were extended.The absorbances in the strong positive,positive, weakly positive and negative samples were detected and analyzed with SPSS 1 7.0 software.Results ①Extending the incubate time before dispending the enzyme had a statistical difference between the strong positive and weakly positive samples.The absor-bances of strong positive samples were obviously decreased while weakly positive samples were improved remarkably,and it was closely related to the time.②Extending the incubate time after dispending the enzyme had the most significant influences on each specimen except the negative one.③Extending the coloration time did not have obvious influence on the experimental results.④Extending the incubate time of different experimental steps had no statistical influence on the absorbance of the HBsAg negative＆nbsp;samples.Conclusion Extending the incubate time of the right steps could improve the accuracy and the detective rate of weakly positive sample in ELISA experiments.