Abstract
|
Research on the Vertical Transmission of Hepatitis C Gene from Father-to-child via Human Sperm
by Mingfu Ma, Yijian Zhu, Dian Wang, Zhiwei Hou, Jihua Huang, Danyan Zhang, Qingdong Xie, Ling Wan, Letian Zhao, Li Wei, Lianbing Li, Tianhua Huang
|
|
|
Background: The aim was to develop a better experimental model which could facilitate further studies assessing the vertical HCV gene transmission via human spermatozoa, and verify the possibility of father-to-child transmission of the HCV gene.
Methods: The recombinant plasmid pIRES2-EGFP-HCV C was constructed. Fluorescence in situ hybridization was performed to detect the integration of the HCV C gene in human sperm genome and in zygote’s pronucleus.
Results: Successful construction of recombinant plasmid pIRES2-EGFP-HCV C was confirmed by restriction mapping, PCR, and sequencing. Positive HCV C DNA signals were observed in sperm heads, human sperm chromosomes and two-cell embryos in transfected samples. No positive signal was found in normal control and HCV infected groups.
Conclusions: The recombinant plasmid pIRES2-EGFP-HCV C was successfully constructed. The HCV C gene was able to pass through the sperm membrane and integrate into the sperm genome. Human sperm carrying the HCV C gene was able to achieve normal fertilization. The replication of the sperm-mediated HCV C gene was synchronized with that of the host genome. Our results provide direct evidence for vertical transmission of the HCV C gene from father-to-child via human sperm.
DOI: 10.7754/Clin.Lab.2015.150706
|
