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目的利用竞争ＰＣＲ法建立分级测定ＨＢＶＤＮＡ含量的定量方法．方法设立３个标准竞争模板（１０２ｃｏｐ，１０４ｃｏｐ，１０６ｃｏｐ），分别和恒量的待测模板混合，分别进行４０，３０，２０次循环的ＰＣＲ扩增，最后凝胶电泳分离待测模板和竞争模板的ＰＣＲ产物，测定两者的荧光强度的比值，计算待测模板的初始量．结果对乙型肝炎血清ＨＢＶＤＮＡ定量表明，１２份ＨＢｅＡｇ阳性血清，ＨＢＶＤＮＡ的血清浓度在６×１０７～１×１０１１ｃｏｐ／Ｌ，而１２份ＨＢｅＡｂ阳性血清只有７份可检出ＨＢＶＤＮＡ，它们的血清浓度均在３×１０８ｃｏｐ／Ｌ以下，其余５份用该ＰＣＲ方法，检测不到．结论该方法可用于ＨＢＶＤＮＡ以及其他病原体ＤＮＡ的定量
Objective To establish a quantitative method for the determination of HBVDNA content by competitive PCR method. Methods Three standard competitive templates (102cop, 104cop, 106cop) were set up, which were respectively mixed with a constant template to be tested. PCR amplification was carried out for 40, 30 and 20 cycles, respectively. Finally, gel electrophoresis was used to separate the template from the competition template PCR products were measured fluorescence intensity ratio of the two to calculate the initial amount of template to be tested. Results The quantitative analysis of serum HBVDNA of hepatitis B showed that the serum concentration of HBVDNA in 12 HBeAg positive sera was 6 × 107 ~ 1 × 1011 cop / L, while only 7 out of 12 HBeAb positive sera could detect HBVDNA, and their serum concentrations Below 3 × 108cop / L, the remaining 5 copies were undetectable by this PCR method. Conclusion This method can be used for quantification of HBVDNA and DNA of other pathogens