Detection of hepatitis B viral DNA integration in HepG2.2.15 cells by a modified Alu-PCR method

Abstract

Abstract: Objective To investigate the integration site of hepatitis B viral DNA (HBV DNA) in HepG2.2.15 cells using a modified Alu-PCR method.Methods This study detected the HBV integration site in HepG2.2.15 cells using a simplified Alu-PCR method, followed by a quantitative analysis of the integration site that was found in HepG2.2.15 cells with and without H₂O₂ treatment by RT-qPCR.Results One HBV integration site was found. The binding junction of the inserted viral fragment was at 1,228nt of HBV DNA with 3 bp (CTG) of microhomology. The viral fragment was inserted into Alu repeats of host DNA in HepG2.2.15 cells. Logarithmically transformed analysis (log10copies/cell) showed that the average copy numbers of this integration site in the cells with H₂O₂ treatment (-1.13±0.07) were significant higher than those without H₂O₂ treatment (-2.10±0.82,P<0.001). No significant difference was found between the HBV cccDNA levels in cells with and without H₂O₂ treatment (-1.94±1.45 and -1.79±1.40,P=0.915). No correlation was found between cccDNA level and the integration site in the cells (P=0.463).Conclusion This study provided a cost-effective, simplified method for the detection of HBV DNA integration by a modified Alu-PCR method.

Key words: Hepatitis B virus, HBV integration, Alu-PCR, HepG2.2.15 cell

RUAN Peng, HE Chun-ping, HUANG Chao, ZHOU Rui. Detection of hepatitis B viral DNA integration in HepG2.2.15 cells by a modified Alu-PCR method[J]. Chinese Hepatolgy, 2022, 27(7): 785-788.

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