改良聚合酶链反应检测HBV共价闭合环状DNA
http://www.100md.com
汤勃, 王宇明, 刘俊, 张瑞
肝炎病毒，乙型;共价闭合环状DNA;聚合酶链反应;绿豆芽核酸酶,汤勃，王宇明，刘俊，张瑞，,汤勃,通讯作者：,DetectionofhepatitisB
 |
| 第1页 |
参见附件(483KB，5页)。
汤勃，王宇明，刘俊，张瑞，第三军医大学西南医院全军感染病研究所 重庆市 400038
汤勃, 男, 1973-04-16生,安徽省宣城市人, 汉族, 2003年第三军医大学博士, 主治医师, 主要从事肝炎病毒分子生物学研究.
国家自然科学基金资助项目，No. 30230320
通讯作者：汤勃, 400038, 重庆市沙坪坝区西南医院感染科.
botang@mail.tmmu.com.cn
电话：023-65461189-8044
收稿日期：2005-07-19 接受日期：2005-07-28
Detection of hepatitis B virus cccDNA with modified polymerase chain reaction
Bo Tang, Yu-Ming Wang, Jun Liu, Rui Zhang
Bo Tang, Yu-Ming Wang, Jun Liu, Rui Zhang, Department of Infectious Diseases, Southwest Hospital of the Third Military Medical University, Chongqing 400038, China
Supported by National Natural Science Foundation of China, No. 30230320
Correspondence to: Bo Tang, Department of Infectious Diseases, Southwest Hospital of the Third Military Medical University, 400038, Chongqing, China. tobei@163.com
Received: 2005-07-19 Accepted: 2005-07-28
Abstract
AIM: To establish a simple and fast hepatitis B virus covalently closed circular DNA(cccDNA) detecting method based on polymerase chain reaction(PCR) with satisfactory sensitivity and specificity.
METHODS: The cccDNA and the relaxed circular DNA (rcDNA) were extracted from HepG2.2.15 cells and supernatant, respectively, and then purified. Two pairs of specific PCR primes were designed to cover the single strand area of rcDNA. And two pairs of non-specific PCR primes were designed to cover the double strand area of rcDNA. Before and after digested by single-strand-specific mung bean nuclease(MBN), cccDNA and rcDNA samples were amplified by specific primes and non-specific primes. Whether the digested cccDNA can be amplified by specific primes, without amplifying the digested rcDNA, was observed. The PCR parameters such as substrate amount and circulation times were changed during amplification. The HBV genome plasmid was used as control; and the HBV samples from patient with hepatitis B was used for practical test ......
您现在查看是摘要介绍页，详见PDF附件(483KB，5页)。