Establishment of microarray for detecting mutation in HBV pre-core/core and basic core promoter regions
10.3760/cma.j.issn.1003-9279.2010.05.024
- VernacularTitle:乙型肝炎病毒核心启动子及前C区突变检测基因芯片的制备及临床应用
- Author:
Li-Juan FANG
1
;
Le-Xiang LAI
;
Yi-Quan LE
;
Ling-Jun REN
Author Information
1. 杭州市萧山区第四人民医院
- Keywords:
Hepatitis B Virus;
Mutation;
Oligonucleotide array sequence analysis
- From:
Chinese Journal of Experimental and Clinical Virology
2010;24(5):389-391
- CountryChina
- Language:Chinese
-
Abstract:
Objective To develop a sensitive and specific microarray for detecting mutations of HBV pre-core/core and basic core promoter regions in the clinic. Methods Site-specific oligonucleotide probes were designed and immobilized to microarray slides and hybridized to HBV gene fragments amplified with specific biotin-labeled primer using asymmetrical PCR. The specificity and sensitivity of the method were estimated. And the microarray was applied to detect 138 clinical serum samples with HBV-DNA.Results The mutations of HBV pre-core/core and basic core promoter regions can be specifically detected using the microarray, and the sensitivity was 1 × 101 copies/μl. Among 138 samples,40 samples had T1762/A1764 mutation, 1 lsamples had C1814 mutation, and 16 samples had A1896 mutation. The A1896 mutation rate in high HBV-DNA load group was significantly higher than that in low HBV-DNA load group (P ＜ 0. 01 ). Conclusion An DNA microarray assay was successfully established to detect the mutations in HBV pre-core/core and basic core promoter regions. The A1896 mutation in Pre-core/core region maybe involve in duplication of HBV.
