Construction of the standards for detecting plasma EBV-DNA in patients with nasopharyngeal carcinoma using real-time fluorescent quantitative polymerase chain reaction

Objective: To construct a series of standards for detecting plasma human herpes virus 4 DNA(EBV-DNA) in patients with nasopharyngeal carcinoma.Methods: According to the sequence data for the EBV genome from Genebank sequence database,the specific PCR primers and the fluorescent probes were designed and synthesized,the specific DNA fragment were amplified by conventional PCR.The products of PCR above were purified and cloned into T-vector named PMD-18 plasmid and the recombined plasmid was transformed into DH5α E coli,and was extracted and from the reproduced E coli.After identifying and purifying the sequences were used as quantitative standard templates for the fluorescent quantitative PCR detecting.Results: The specific DNA sequence was recombined with PMD-18T vector was proved by PCR amplification and sequence analysis.Conclusions: We established the standards for detecting the EBVDNA using fluorescent quantitative PCR successfully.