摘要
目的验证乙型肝炎（简称乙肝）5项酶联免疫吸附试验（ELISA）定性检测的分析性能,改进和优化试验方案,为实验室选用合适的方法和试剂提供依据。方法通过梯度稀释定值样品测定检出限;依据美国临床实验室标准化协会（CLSI）EP12-A2文件进行临界值验证;选取临界值样品评价分析精密度（包括重复性和中间精密度）;通过与室间质评检测结果及雅培i4000SR化学发光仪比对评估符合率。结果乙肝表面抗原（HBsAg）、乙肝表面抗体（HBsAb）、乙肝e抗原（HBeAg）、乙肝e抗体（HBeAb）及乙肝核心抗原（HBcAb）的最低检出限分别为：0.2IU/mL、20mIU/mL、1NCU/mL、0.75NCU/mL、0.05NCU/mL。临界值（C50）±20%的浓度范围包含了多次重复试验阳性率5%~95%的浓度点区间。批内变异系数（CV）≤15%,夹心法批间CV≤25%,竞争法批间CV≤35%。准确度及与雅培i4000SR比对的阴、阳性符合率均大于或等于95%,其一致性检验κ≥0.75。结论本实验室乙肝5项ELISA检测方法性能符合检测分析要求,检测质量可满足临床需求。
Objective To evaluate the performance of enzyme linked immunosorbent assay（ELISA）kit in detection of Hepatitis B virus（HBV）markers by using improved and optimized method,so as to provide a practical and feasible method and reagents for clinical laboratory.Methods ELISA test was used for the detection of HBV markers.The gradient dilution method was used to evaluate the lower limit.The verifiation of cut off value was carried out based on clinical and laboratory standards institute（CLSI）EP12-A2 document.Samples with cut off values were collected to evaluate the precision,including repeatability and intermediate precision.The coincidence rates were counted through comparing the results of ELISA with those of external quality assessment and those detected by using Abbott i4000 SR chemiluminescence instrument.Results The lower detection limit of HBsAg,HBsAb,HBeAg,HBeAb and HBcAb were 0.2IU/mL,20mIU/mL,1NCU/mL,0.75NCU/mL and 0.05NCU/mL respectively.The cutoff value（C50）±20% concentration included the concentration range between C5 and C95.The within-run coefficient of variation（CV）≤15%,in sandwich method the between-run CV≤25%,in competition method the between-run CV≤35%.The positive and negative coincidence rates in accuracy and comparing with i4000 SR all were more than 95% and allκ0.75.Conclusion ELISA tests for HBV markers in our laboratory could meet the requirements of the detection performance and clinical needs.
出处
《国际检验医学杂志》
CAS
2015年第23期3369-3371,共3页
International Journal of Laboratory Medicine
基金
国家临床重点专科建设项目经费资助项目[财社(2010)305号]
重庆市渝中区科技计划项目(20140120)
关键词
乙型肝炎标志物
酶联免疫吸附试验
性能验证
hepatitis B markers
enzyme linked immunosorbent assay
performance verification
