摘要
目的了解肝病患者血清乙型肝炎病毒（ＨＢＶ）ＤＮＡ的含量，以便观察临床抗病毒治疗效果及血清ＨＢＶＤＮＡ水平与病情及预后的关系。方法应用荧光素标记的半巢式引物在扩增中能量转移的定量聚合酶链反应（ＱＰＣＲ），对６３例肝功能异常的肝病患者血清进行ＨＢＶＤＮＡ含量测定，并与普通ＰＣＲ及酶联免疫吸附试验（ＥＬＩＳＡ）进行比较。结果ＱＰＣＲ阳性率为８２５４％，普通ＰＣＲ法为７１４３％，ＥＬＩＳＡ法为６３５０％。３０例健康人血清ＱＰＣＲ检测ＨＢＶＤＮＡ均为阴性。ＱＰＣＲ与普通ＰＣＲ的符合率为８８８９％，Ｐ＜００５；与ＥＬＩＳＡ的符合率为８０９５％，Ｐ＜００５。ＱＰＣＲ检测血清中ＨＢＶＤＮＡ平均含量（ｘ±ｓ）与ＥＬＩＳＡ－ＨＢＶ血清平均滴度比较，ＱＰＣＲ与ＥＬＩＳＡ均阳性组血清ＨＢＶＤＮＡ含量明显高于ＱＰＣＲ阳性、ＥＬＩＳＡ阴性组，经ｔ检验，Ｐ＜００５。结论ＱＰＣＲ方法具有特异性强、敏感度高，可检出低水平ＨＢＶＤＮＡ等优点。
In order to test the content of serum HBV DNA for evaluating the effectiveness of clinical antiviral treatment and the relation of serum HBV DNA level to state of illness and prognosis, a QPCR method of energy transfer in amplification by semi-nested fluorescein labeled primer was established. Serum HBV DNA contents of 63 cases of hepatic diseases with abnormal liver function were detected by QPCR, PCR and ELISA. The positive detectable rate of the three methods were compared. The results showed that the positive detectable rate of QPCR was 82.54%, PCR 71.43% and ELISA 63.50%. The detectable rate of QPCR was compared with PCR, and uniformity rate of the both methods was 88.89%, P<0.05; the positive results of QPCR were compared with ELISA, and their uniformity rate was 80.95%, P<0.05; when serum average content (±s)of HBV DNA by QPCR method was compared with serum average titer of HBV by ELISA method, serum HBV-DNA average content of the group in which both QPCR and ELISA were positive was much higher than that of the group of QPCR(+) and ELISA(-), P<0.05; serum QPCR-HBV-DNA of 30 cases of healthy persons were all negative. The results indicated that QPCR method is of highest positive detectable rate and more specific in detecing serum HBV DNA and had no nonspecific amplifying reaction. It has the superiority of detecting low level serum HBV DNA.
出处
《中华实验和临床病毒学杂志》
CSCD
1998年第4期367-369,共3页
Chinese Journal of Experimental and Clinical Virology
