Integration sites in HCC biopsy
10.3760/cma.j.issn.1003-9279.2010.05.023
- VernacularTitle:三种PCR方法在HBV基因整合位点确定中的应用比较
- Author:
Lan-Tian WANG
1
;
Bao-Hua ZHU
;
Bo-Ping ZHOU
;
Wei-Long LIU
;
Ming-Feng LANO
;
Xiao-He LI
;
Xin-Chun CHEN
;
Tao LI
Author Information
1. 广东医学院第一临床学院
- Keywords:
Polymerase chain reaction;
Hepatitis B virus;
Carcinoma,hepato cellular;
Virus integration
- From:
Chinese Journal of Experimental and Clinical Virology
2010;24(5):386-388
- CountryChina
- Language:Chinese
-
Abstract:
Objective To compare the performance of Inverse-PCR, Alu-PCR and Cassetteligation-mediated PCR (CLM-PCR) in HBV DNA integration sites identification. Mehtods One HCC biopsy was obtained from surgically resected sample. The patient was positive for serum hepatitis B surface antigen (HBsAg). The genomic DNA was purified by the standard phenol/chloroform extraction and ethanol precipitation method. Seperated set of primers were designed to amplify the HBV DNA integration region by means of 3 different PCR methods respectively. The PCR products were analyzed by electrophoresis, then cloned to PMD18-T vector for DNA sequencing. The sequence alignment was performed under Blast software. Results 7 bands and 22 sequencing results was obtained from IPCR and 3 integration sites was identified. Alu-PCR provided 12 bands and 32 sequencing results, and CLM-PCR showed 12 bands and 4sequencing results. No integration site was identified from the latter two. Conclution IPCR compared with another two methods showed a reliable capacity in HBV DNA integration site identification.
