乙型肝炎病毒全基因组pcr扩增产物的直接测序.pdf

20 () 2009 2 3 1 Ch ni J Exp Clin InfectDis (Electron icEditoin), Februar 2009, Vol 3, No. 1 PCR 陈伟烈 蔡晓莉 魏绍静 杨湛 唐小平 PCR PCR , PCR DNA ; PCR ; HBV YMDD 20 18 ; , 1; , 18 10 B8 C; 2 YM DD, C PCR , , ; PCR; D irect sequencing of PCR am plification products of Hepatitis B virus complete genom e CHEN Weilie, CAI X iaoli, WEI Shaoj ing, YANG Zhan, TAN G X iao p ing. Institute of Inf ectious D iseases, the th Peop lesH osp ital of Guangzhou, Guang zhou 510060, China Corresp onding author: CHEN Weilie, Email: cw @l tom. com Abstract Objective To establish a method for direct sequencing the PCR amplification products of complete genome of hepatitis B virus and anal se the se quences. M ethods The complete genome of HBV w ere amplified b PCR. PCR products w ere sequenced directl. PCR products w ere resequenced to evaluate the fi delit of the m ethod. The genot pe and YM DD mutation of HBV were determ ined b ph logenetic anal sis and alignm ent. Results E ighteen complete genome of HBV were amplified b PCR from 20 samples and the genome were sequenced; the fidelit anal sis show ed that artificial mutation rate of the method w as 1. In 18 samples which HBV complete genom e were amplified, 10 cases were genot pe B, 8 cases were genot pe C, 2 cases w ere YM DD mutation and both w ere genot pe C. Conclusions The PCR amplification products of HBV complete genom e can be : ( 2004Z008) : 510060 , : , Em ail: cw l@ tom. com ()2009 2 3 1 Ch ni J Exp Clin InfectDis (Electronic Edition), Februar 2009, Vo l3, No. 1 21 sequenced directl , th is m ethod is smi ple and conven ient which can be used in se quence anal sis ofHBV complete genom e on a large application. Ke words Hepatitis B virus; PCR products; Sequencing DNA , 3200, , : C /C PSX , , PCR , , HBV HBV