【作者】 王蔚青； 王彤； 孟玲； 杜青； 周思蒙；

【Author】 WANG Wei-qing 1,WANG Tong 1,MENG Ling 1,DU Qing 1,ZHOU Si-meng 2 1The First Affiliated Hospital of Nanjing Medical University,Nanjing 210029; 2University Paul Cezanne-Aix-Marseille 3,13003

【机构】 南京医科大学第一附属医院； 艾克斯马赛第三大学；

【摘要】 目的:建立实时定量PCR法测定血清样品中乙肝病毒脱氧核糖核酸(HBVDNA)含量。方法:用PEG-裂解-煮沸法从血清中提取核酸,制作成标准浓度曲线后,对100份患者样本进行实时荧光定量分析,并与ELISA(酶标法)结果作比较,再与CAP-CTM法进行比较。结果:用SPSS(v.16)软件进行统计分析,线性回归方程Y=-3.212X+44.90;r2=0.9980;日内、日间平均误差为RSD<2.15%;最低检测限是400copies/ml(拷贝数);该法测得患者HBVDNA的浓度与ELISA测出样品阳性发生率呈正相关,再与CAP-CTM(内标法)作配对t检验,P>0.05,二者无显著性差异。结论:新建的PCR检测法敏感度高,重现性好,线性范围较广,可用于进行临床血清样本中HBVDNA的定量检测,这对评估HBV患者诊治效果有帮助。更多还原

【Abstract】 Objective:To establish a real-time PCR method for the determine of HBV DNA concentration in the human serum. Methods:We utilized PEG-lyses-boiling method extracting HBV DNA from in the serum sample, these samples (concentration of seven pieces) were used to determine the standard linearty of the assay. To assay 100 cases of serum with the real-time PCR, result was done between two kinds of analysis methods to by comparison. Results:Statistical analysis was done with SPSS statistical of software(version16); A good linearty was seen the calibration curves when the value r 2 =0.9980.Linear equation of regression was Y=-3.212X+44.90. The intra-day and inter-day precision(RSD) was<2.15%. The minimum detection limit was 400 copies/ml. Quantitive assay of the real-time PCR has positive relevance and HBV DNA positive rate of ELISA test, but show no significant difference in the CAP-CTM test from the latter(P>0.05). Conclusions:This method can be used for the determination of the HBV DNA concentration in the human serums and for the value of the HBV patient of short-term efficacy.更多还原