摘要
目的乙型肝炎病毒PreS1抗原时间分辨免疫荧光分析方法临床应用研究。方法应用双抗体夹心免疫荧光分析方法,用抗PreS1单克隆抗体和抗-HBs作为固相抗体和铕标记抗体,若样本中存在PreS1抗原,则形成抗体-抗原-铕标记抗体复合物,加增强液解离铕离子,采用时间分辨荧光测量技术,对样本中乙型肝炎病毒PreS1抗原进行检测。结果自制TRFIA试剂与ELISA试剂对300例乙型肝炎患者血清标本中PreS1抗原的检测,TRFIA方法更灵敏,有9例标本ELISA方法结果为阴性,而用TRFIA方法可检测到PreS1抗原。对这9例标本进行HBV DNA定量检测,有6例标本HBV DNA拷贝数均>106;高、中、低三个浓度,TRFIA方法批内和批间精密度测试,CV均小于10%;TRFIA方法从低值到高值,从批内至批间CV均优于ELISA方法;对于强阳性标本,从原倍到1∶16倍稀释,ELISA方法都无法区分阳性程度的强弱,而TRFIA方法结果从高到低有显著性区别。对于弱阳性标本用ELISA方法检测不到时,TRFIA方法仍可检出,一强阳性标本,ELISA方法在1∶8192倍稀释时结果为阴性,而TRFIA方法在1∶16384倍稀释时仍可检出PreS1抗原。结论TRFIA方法与ELISA方法相比,测量范围、精密度和灵敏度有较大提高。
Objective Clinical application of time-resolved fluoroimmunoassay for the detection of hepatitis B virus （HBV） PreS1 antigen. Methods Firstly the monoelonal antibody of HBV PreS1 antigen was covered on the microwell plate incubated with HBV PreS1 antigen in sample for an hour at room temperature. Secondly rinse the wells, HBV PreS1 incubated with Eu3 ＋ labled Hepatitis B surface antibody an hour at room temperature. Thirdly rinse the wells . Next the Eu3 ＋ was released by adding enhancement solution . At last to detect HBV PreS1 antigen in sample with time-resolved fluoroimmunoassay instrument. Results The method of TRFIA is more sensitive than ELISA. Tile intra- and inter-assay coefficients of variation were less than 10% in high, medium and low densities.The ELISA can＇t distinguish the strong positive degree from original times to it＇s 1：16 dilutions of a strong positive specimen but the positive degree can be distinguished by using T RFIA. TRFIA can detect PreS1 antigen in a strong positive specimen with 1：16384 dilutions,but the PreS1 antigen can not be detected using the ELISA method.Conclusion TRFIA method has more widely measure scope and more sensitivie than ELISA for the detection of PreS1.
出处
《中国实验诊断学》
2007年第12期1641-1644,共4页
Chinese Journal of Laboratory Diagnosis
