Study on amplification of the nucleotide sequences homologous to GBV-C from the DNAs of MT2 and HeLa cells and the PBMC DNAs of human and chimpanzee
- VernacularTitle:从MT2、HeLa细胞DNA及人和黑猩猩PBMC DNA中扩增GBV-C核苷酸
- Publication Type:Journal Article
- From: Chinese Journal of Microbiology and Immunology 2001;21(2):221-224
- CountryChina
- Language:Chinese
- Abstract: Objective To detect the nucleotide sequences homologous to GBV-C existed in the DNAs from MT2 and HeLa cells and the PBMC DNAs from 6 human samples and 1 chimpanzee sample. Methods Direct nested-PCR (dPCR) respectively amplified with GBV-C 5′-NCR and NS3 primers by using the DNAs as templates, nucleotide sequence analysis, primed in situ (PRINS) sequence-specific labeling with fluorescein. Results DNA fragments amplified with GBV-C 5′-NCR primers were obtained from the DNAs of MT2 and HeLa cells and the PBMC DNAs in 4 of 6 human samples. DNA fragments amplified with GBV-C NS3 primers were obtained from MT2 and HeLa cells and the PBMC DNAs in 5 of 6 cases of human cases and chimpanzee. The homology of the nucleotide sequences of these amplification products compared with GBV-C genomic sequence were 74.29%-77.14% and 73.80%-79.15% respectively. The fluorescence staining by PRINS was shown in PBMCs and their chromosomes with positive PCR results. Conclusion The nucleotide sequences with higher homology to GBV-C 5′-NCR and/or NS3 region existed in the DNAs from MT2 and HeLa cells and in the PBMC DNAs of human and chimpanzee. The sequences of the PBMCs with positive PCR results were expected to locate in their chromosomes.