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| Expression of Rta Recombinant Antigen of Epstein-Barr Virus and Establishment of ELISA Method for Nasopharyngeal Carcinoma Screening |
| ZHENG Fengjiao, FU Yonghang, CHEN Wensi, LIU Nan* |
| Department of Clinical Laboratory, Hospital 458 of PLA, Guangzhou 510602,. China |
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Abstract Objective To prepare and express the Rta recombinant antigen of Epstein-Barr virus and to establish an ELISA method in order to facilitate the screening of nasopharyngeal carcinoma (NPC).Methods The target gene BRLF1 which coded for Rta was ampliflied by PCR with a specific primer. The recombinant vector pPICZαA-BRLF1 was electronically transformed into pichia pastoris GS115 to express Rta protein so as to screen the His+Mut+ phenotype transformant before proteins were purified and the immunogenicity of the product was proved by Western blot. ELISA was established with the IgG/Rta antibody and was applied to the detection of 300 patients with NPC and 200 healthy people to assess the capability of detection.Results The pPICZαA-BRLF1 recombinant plasmid was successfully constructed and the target protein was secreted and expressed with high-performance (50%). Protein electrophoresis demonstrated a corresponding band in 66 KDa. Western blot showed a good immune-competence with the IgG monoclonal antibody of Rta. The Rta recombinant antigen was used for establishment of the ELISA method. The positive rate of this method was significantly higher than that of IgA/VCA Kit for screening NPC (P<0.01). Conclusion A eukaryotic expression vector of the Rta recombinant antigen of Epstein-Barr virus has been constructed that can ensure the high-performance of foreign proteins with good immunogenicity for the development of IgG/Rta ELISA methods. The detection capability can be improved by combining other antigens and this method is applicable to large-scale screening and diagnosis of NPC.
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Published: 14 September 2016
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2016, Vol. 34 
