摘要
目的对高敏乙型肝炎(乙肝)病毒脱氧核糖核酸(HBV DNA)定量检测进行方法学评价,并探讨该方法在低病毒载量的乙肝患者治疗监测中的应用价值。方法采用全自动核酸提纯及荧光聚合酶链式反应分析系统检测HBV DNA,评价该方法的线性范围、精密度、最低定量检测限、防污染能力和最低检测限等性能指标,采用电化学发光法检测HBV血清学标志物和全自动生化仪检测丙氨酸氨基转移酶(ALT),并分析两者之间的相互关系。结果高敏HBV DNA定量检测方法的线性范围为50~5.0×10~8 IU/ml,最低定量检测限为50 IU/ml,批内精密度与批间精密度均<5%,最低检测限为20 IU/ml,防污染能力强。在抗病毒治疗的中,HBV DNA载量<500 IU/ml的乙肝患者占总数的46.40%;HBVDNA载量为30~500 IU/ml的HBeAg阳性或阴性患者的ALT异常率明显高于HBV DNA载量<30 IU/ml的患者(P=0.012、0.001),但不同HBV DNA载量与两对半血清学模式差异无统计学意义(P=0.179)。结论基于全自动核酸提纯及荧光PCR分析系统的高敏HBV DNA定量检测方法各性能指标良好,适于低病毒载量的乙肝患者治疗监测,临床应加强该类乙肝患者的治疗监测。
Objective To evaluate the performance of highly sensitive HBV DNA quantitative detection and discuss its application in the treatment monitoring of hepatitis B patients with low viral load. Methods Automatic nucleic acid extraction and fluorescence PCR analysis system was used to detect HBV DNA, the linear range, precision, limit of detection （LOD） , limit of quantitative detection and anti-pollution capability of the method were evaluated. HBV serological markers were detected by electrochemiluminescence immunoassay. ALT was detected by automatic biochemical analyzer. And the relationship between HBV DNA, serological markers and ALT were analyzed. Results The result showed that the linear range of high sensitivity HBV DNA quantitative detection was between 50 IU/ml and 5.0x 108 IU/ml. The LOD of the method reached 30 IU/ml. The precision of inter and intra-run CV were all less than 5%. And the method had a strong anti- pollution capability. The patients with HBV DNA load 〈500 IU/ml accounted for 46.4%. In HBeAg positive or negative group, the ALT abnormal rate of patients with HBV DNA load ranging from 30 IU/ml to 500 IU/ml was significantly higher than those with HBV DNA load 〈30 IU/ml （P=-0.012, P=-0.001） , but different HBV DNA load had no relationship with serological markers patterns （P=0.179） . Conclusion Automatic nucleic acid extraction and fluorescence PCR analysis system based highly sensitive HBV DNA quantitative detection has good performance, which is suitable for the treatment monitoring of patients with low hepatitis B viral load. And the treatment monitoring of such hepatitis B patients should be enhanced.
作者
金速速
陈占国
余坚
白伟伟
陈碎朋
谢振迪
李向阳
Jin Susu, Chen Zhanguo, Yu Jian, Bai Weiwei, Chen Suipeng, Xie Zhendi, Li Xiangyang.(Department of Laboratory Medicine, The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, Zhejiang, Chin)
出处
《疾病监测》
CAS
2018年第1期67-71,共5页
Disease Surveillance
基金
浙江省医药卫生科技计划(No.2017KY112)
温州市公益性科技计划(No.Y20160161)~~
关键词
高敏
乙型肝炎病毒
核酸
性能验证
低载量
High sensitivity
Hepatitis B virus
Nucleic acid
Performance verification
Low load
