实时荧光定量PCR检测血HBV DNA的疑难结果分析及其对策  

Analysis of suspicious results of serum HBV DNA detected by fluorescence quantitative PCR

作　　者：陈占国[1] 周武[1] 王忠永[1] 金娅蕾[2] 陶志华[3] 

机构地区：[1]温州医学院附属第一医院实验诊断中心,温州325000 [2]温州医学院温州市第三临床学院 [3]浙江大学医学院附属第二医院检验科

出　　处：《中华检验医学杂志》2013年第3期217-221,共5页Chinese Journal of Laboratory Medicine

基　　金：国家自然科学基金资助项目（30872421）;温州市科技局资助项目（Y20090280）

摘　　要：目的分析实时荧光定量PCR检测血HBV DNA过程中出现的疑难结果，并探讨制定相应的处理对策，以提高HBV DNA检测质量。方法用实时荧光定量PCR检测温州医学院附属第一医院2008至2011年期间，115154份血标本中HBV DNA含量，结合当次HBV DNA定量结果与历史结果、PCR扩增图、乙型肝炎血清5项指标及临床诊断，建立本实验室疑难结果复查标准，回顾性分析2008至2011年本实验室所有送检标本的疑难结果，同时用原试剂（careHBV PCR Kit）和验证复查试剂（careHBV PCR Kit V2）对1969份疑难结果标本进行复查；统计2次结果一致性和不一致的比例，并分析试剂与非试剂因素等造成2次结果不一致的比例，其中试剂因素包括无扩增曲线造成的假阴性和有扩增曲线但扩增效率低2种情况，非试剂因素包括操作污染、标本等其他因素；最后统计careHBV PCR Kit的漏检率。结果2008至2011年共定量检测115154份HBV DNA血标本，疑难结果标本1969份，占总检测标本的1．71％；2次结果一致的为1588份（80．65％）；2次结果不一致为381份（19．35％），其中，4年中试剂因素造成结果不一致的比例分别为18．87％、20．23％、51．33％和59．57％，呈逐年上升趋势，原试剂因素的漏检率分别为2．49％、4．08％、10．09％、14．47％，呈逐年上升趋势；4年中非试剂因素造成结果不一致所占比率分别为81．13％、79．77％、48．67％和40．43％，呈逐年下降趋势。结论用2种不同的试剂复查疑难结果标本，可降低HBV DNA定量检测的漏检率，避免试验误差，有效保证HBV基因突变标本的HBV DNA准确定量，以正确指导乙型肝炎患者的抗病毒治疴0（中华检验医学杂志，2013，36：217-221）Objective To analyze the suspicious results of serum HBV DNA by fluorescence quantitative PCR and develop appropriate countermeasures in order to improve the quality of detection of HBV DNA. Methods Blood samples of patients from the First Affiliated Hospital of Wenzhou Medical College from 2008 to 2011 were analyzed for HBV DNA by fluorescence quantitative PCR. 1969 cases of suspicious results, judged by the rule of review the results of serum HBV DNA combined with the historical results, PCR amplification curve, HBV serum markers and clinical diagnosis, were analyzed and redetected by using of two different reagents, careHBV PCR Kit and careHBV PCR Kit V2, at the same time. The consistency and inconsistency ratio of the results were evaluated. Both the reasons of inconsistent and the undetected rates of careHBV PCR Kit were analyzed. The two reasons for the inconsistent resuhs included the reagent related factors, e. g, showing no amplification curve caused by the false negative and abnormal low efficiency of amplification curve, and the non reagent related factors such as operating pollution and other sample factors. Results There were 115 154 blood samples were detected for HBV from 2008 to 2011 and 1969 samples （1.71%） with suspicious results were redetected. The consistency and inconsistency results were 1588 （80. 65% ） and 381 （ 19. 35% ）, respectively. Every year from 2008 to 2011, the percentage of the inconsistent results caused by the reagent related factors were 18.87% , 20. 23% , 51.33% and 59. 57% respectively, which showed an increasing trend, and the percentage of inconsistent resuhs caused by the nonreagent related factors were 81.13%, 79.77%, 48.67% and 40. 43% respectively, which showed a declining trend year by year. The undetected rates of careHBV PCR Kit were 2. 49%, 4. 08%, 10. 09% and 14.47% respectively, showing an increasing trend. Conclusions The redetection for the specimens with the suspicious results by using of different reagents can avoid the blind detection of HBV DNA

关 键 词：聚合酶链反应 肝炎病毒 乙型 DNA 病毒 

分 类 号：R512.62[医药卫生—内科学]