乙型肝炎病毒DNA的锁核酸实时荧光定量PCR探针法检测
LNA real-time PCR probe quantification of hepatitis B virus DNA.
作者信息
Wang Qing, Wang Xueqian, Zhang Junhua, Song Guanghui
机构信息
Department of Clinical Laboratory, The Affiliated Hospital of Qingdao University Medical College;
出版信息
Exp Ther Med. 2012 Mar;3(3):503-508. doi: 10.3892/etm.2011.442. Epub 2011 Dec 28.
In the present study, we standardized a TaqMan locked nucleic acid (LNA) real-time polymerase chain reaction (PCR) probe for the accurate quantification and detection of hepatitis B virus (HBV) DNA in serum (plasma), and evaluated its methodology. LNA probe technology had a much better detection performance in HBV DNA than the common TaqMan probe. The assay based on the LNA probe had a wider linear detection range, higher sensitivity, stability and amplification efficiency, and a lower concentration of probes than the TaqMan probe. Among the 15 cases with chronic hepatitis B surface antigen (HBsAg) (+) alone, only 4 cases that were detected by TaqMan real-time PCR were negative; however, the same samples were positive by LNA real-time PCR (p<0.05). A positive correlation between viral load measurements for the 35 samples with HBV-positive DNA was detected in both LNA and TaqMan real-time PCR.
在本研究中，我们标准化了一种TaqMan锁核酸（LNA）实时聚合酶链反应（PCR）探针，用于准确定量和检测血清（血浆）中的乙型肝炎病毒（HBV）DNA，并评估了其方法学。LNA探针技术在HBV DNA检测方面比普通TaqMan探针具有更好的检测性能。基于LNA探针的检测方法具有更宽的线性检测范围、更高的灵敏度、稳定性和扩增效率，且探针浓度低于TaqMan探针。在仅慢性乙型肝炎表面抗原（HBsAg）（+）的15例病例中，仅4例通过TaqMan实时PCR检测为阴性；然而，相同样本通过LNA实时PCR检测为阳性（p<0.05）。在LNA和TaqMan实时PCR中均检测到35例HBV DNA阳性样本的病毒载量测量值之间存在正相关。
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