高病毒载量血清HBV DNA定量方法的评价  

Evaluation for quantitative methods of HBV DNA in serum with very high viral loads

作　　者：李雷[1] 冯振华[2,3] 许碧云[4] 顾光煜[1] 张葵[1] 周乙华[2,5,3] 

机构地区：[1]南京大学医学院附属鼓楼医院检验科,南京210008 [2]南京大学医学院附属鼓楼医院科研部,南京210008 [3]江苏省医学分子技术重点实验室,南京210008 [4]南京大学医学院附属鼓楼医院科技处,南京210008 [5]南京大学医学院附属鼓楼医院感染科,南京210008

出　　处：《临床检验杂志》2012年第11期858-861,共4页Chinese Journal of Clinical Laboratory Science

基　　金：南京市卫生青年人才工程第一层次项目(2011024)

摘　　要：目的探讨血清HBV DNA定量检测结果高于检测上限时,能否直接延伸标准曲线,用于HBV DNA定量分析。方法以上海申友生物公司HBV DNA荧光定量检测试剂盒为例,选择HBV DNA>3×107IU/mL的血清标本30例。每例血清在3个不同时间点提取DNA,或将血清以10、100和1 000倍稀释后提取DNA,或将未稀释血清提取的DNA以10、100和1 000倍稀释后进行HBV DNA定量检测,检测结果以10为底数进行对数转换后进行统计学分析。结果经对数转换,3个不同时间点HBV DNA定量检测结果的组间相关系数为0.356(F=2.66,P<0.01);36.7%(11/30)的标本最大差值<0.5,60.0%(18/30)介于0.5～1.0,3.3%(1/30)达1.01;血清稀释后定量检测HBV DNA,96.7%(29/30)的标本最大差值为0.02～0.45,均<0.5,3.3%(1/30)为0.53,介于0.5～1.0;组内相关系数为0.960(F=72.57,P<0.01)。稀释DNA定量检测HBVDNA,最大差值为0.02～0.44,均<0.5;组内相关系数达0.973(F=107.66,P<0.01)。3组均数经方差分析,F=1.66,P>0.05,差异无统计学意义,且43.3%(13/30)的标本的最大差值为0.15～0.48,均<0.5;56.7%(17/30)为0.5～0.85,均介于0.5～1.0。结论定量检测HBV DNA水平超过检测上限时,直接延伸标准曲线可获得较准确的HBV DNA定量结果;但若需准确定量,最好用原倍血清提取DNA经100倍稀释后检测。Objective To explore whether quantitative standard curve may be directly extended to determine HBV DNA level in the cases of very high viral loads which was over the upper limit of detection range. Methods A total of 30 serum samples were included. The HBV DNA levels of all the samples were more than 3 × 10^7 IU/ml detected by fluorescent quantification kit purchased from Shanghai Shenyou Company. Each samples was quantitatively retested for three times. Both the serum samples and the HBV DNA extracted from serum were further measured in 10-, 100- and 1 000-fold dilution, respectively. After logarithmic conversion the quantitative values were statistically analyzed. Results When the quantitative data of HBV DNA were expressed as logarithm values, the inter-class correlation coefficient was 0.356 （ F = 2.66, P 〈 0.01 ） in three repeated detection of the 30 sera. The maximum differences of HBV DNA levels in the repeated detection were 〈0.5, 0.5 to 1 and 1.01 in 11 （36.7%）, 18 （60.0%）, and 1 （3.3%） of the 30 samples respectively. , respectively. After the serum was diluted with 10-, 100- and 1 000-fold, the intra-class correlation coefficient was 0. 960 （ F = 72.57, P 〈 0.01 ）, with the maximum difference of 〈 0.5 （0.02 to 0.45 ） for all the samples except one （0.53）. After the extracted DNA from serum was diluted with 10-, 100- and 1 000-fold, the intra-class correlation coefficient was 0.973 （ F = 107.66 ,P 〈 0.01 ）, with the maximum difference of 〈0.5 （0.02 to 0.44） for all the samples. Comparison of HBV DNA levels in the undiluted samples and diluted with two different methods showed that the averages of the detected results were similar （ F = 1.66, P 〉 0.05）, and the maximum differences were less than 1 in each sample. Conclusion In the determination of HBV DNA by using domestic reagents, direct extension of the quantitative standard curve may be acceptable even if the level of HBV DNA was over the upper limit of detection. For accurate quantification, it i

关 键 词：乙型肝炎病毒 高病毒载量 HBV DNA 定量 

分 类 号：R373.2[医药卫生—病原生物学] R446[医药卫生—基础医学]