通过实时聚合酶链反应快速定量精液中的乙型肝炎病毒DNA
Rapid quantification of semen hepatitis B virus DNA by real-time polymerase chain reaction.
作者信息
Qian Wei-Ping, Tan Yue-Qiu, Chen Ying, Peng Ying, Li Zhi, Lu Guang-Xiu, Lin Marie-C, Kung Hsiang-Fu, He Ming-Ling, Shing Li-Ka
机构信息
The Center for Emerging Infectious Diseases, The Chinese University of Hong Kong, Hong Kong Special Administrative Region, China.
出版信息
World J Gastroenterol. 2005 Sep 14;11(34):5385-9. doi: 10.3748/wjg.v11.i34.5385.
AIM
To examine the sensitivity and accuracy of real-time polymerase chain reaction (PCR) for the quantification of hepatitis B virus (HBV) DNA in semen.
METHODS
Hepatitis B viral DNA was isolated from HBV carriers' semen and sera using phenol extraction method and QIAamp DNA blood mini kit (Qiagen, Germany). HBV DNA was detected by conventional PCR and quantified by TaqMan technology-based real-time PCR (quantitative polymerase chain reaction (qPCR)). The detection threshold was 200 copies of HBV DNA for conventional PCR and 10 copies of HBV DNA for real time PCR per reaction.
RESULTS
Both methods of phenol extraction and QIAamp DNA blood mini kit were suitable for isolating HBV DNA from semen. The value of the detection thresholds was 500 copies of HBV DNA per mL in the semen. The viral loads were 7.5X10(7) and 1.67X10(7) copies of HBV DNA per mL in two HBV infected patients' sera, while 2.14X10(5) and 3.02X10(5) copies of HBV DNA per mL in the semen.
CONCLUSION
Real-time PCR is a more sensitive and accurate method to detect and quantify HBV DNA in the semen.
目的
检测实时聚合酶链反应（PCR）定量精液中乙型肝炎病毒（HBV）DNA的敏感性和准确性。
方法
采用酚提取法和QIAamp DNA血液微量试剂盒（德国Qiagen公司）从HBV携带者的精液和血清中分离乙型肝炎病毒DNA。通过常规PCR检测HBV DNA，并采用基于TaqMan技术的实时PCR（定量聚合酶链反应（qPCR））进行定量。常规PCR的检测阈值为每个反应200拷贝的HBV DNA，实时PCR为10拷贝的HBV DNA。
结果
酚提取法和QIAamp DNA血液微量试剂盒这两种方法均适用于从精液中分离HBV DNA。精液中HBV DNA的检测阈值为每毫升500拷贝。两名HBV感染患者血清中的病毒载量分别为每毫升7.5×10⁷和1.67×10⁷拷贝的HBV DNA，而精液中分别为每毫升2.14×10⁵和3.02×10⁵拷贝的HBV DNA。
结论
实时PCR是检测和定量精液中HBV DNA更敏感、准确的方法。
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