Aim: Since molecular diagnostic methods are expensive and not suitable for the diagnosis of asymptomatic infections, our goal is to obtain a recombinant DNA containing the HPV 18 LI gene. The expressed recombinant protein could act as an antigen in a diagnostic test.
Partners: Coordinator: „Stefan S. Nicolau” Institute of Virology, project director Adriana Plesa, PhD
Working team: Plesa A, Anton G, Repanovici R, Stoian M
Funding: 1 200 000 LEI
Project financed by VIASAN contract no 006
Implementation period: 22.10.2001-31.12.2003
Summary: The assessment of antibody responses against HPVs (previously hampered by the lack of viral source) was enabled by the expression of the LI major capsid viral protein type 18 (HPV 18) into L929 murine cells using the pTARGET mammalian expression vector system (MEVS). The cloning was validated by PCR with specific primers for the LI gene, as well as by enzyme restriction and in situ hybridization. The evidence for the viral cloned gene expression was acquired by RT-PCR. Presence and antigenic properties of the recombinant LI protein were shown using it as antigen in an indirect enzyme linked immunosorbent assay (ELISA) system. Significantly higher reactivity was noted when the sera samples were from persons infected with HPV 18 as compared with the non-infected individuals but a moderately different reactivity was observed when the sera from patients infected with other HPV genotypes were tested. The results showed that the murine transfected cells could be used as antigen in order to delect the presence of the specific antibodies in HPV infected persons.
Results:
- Adriana Plesa, Gabriela Anton, Carmen Diaconu, Mihai Stoian, Rodica Repanovici, Costin Cernescu. Cloning and expression of the L1 major capsid protein of the human papillomavirus 18 in murine cells. International Journal of Molecular Medicine. 2003; 12(6): 1021-1026. https://doi.org/10.3892/ijmm.12.6.1021. IP=5.314/2021
- Adriana Plesa, Carmen Diaconu, Gabriela Anton, Rodica Repanovici. ELISA indirecta pentru detectia anticorpilor neutralizanti din serul unor pacienti, utilizand antigenul viral recombinat L1 HPV18. Stud. Cerc. Virusologie 32, 34-39, 2002.
- Adriana Plesa, Carmen Diaconu, Gabriela Anton, Rodica Repanovici. Detection of anti-L1 HPV antibodies using a recombinant viral antigen. 3rd International Conference of the Chemical Societies of the South-Eastern European Countries on Chemistry in the new millennium-an endless frontier. Sept 22-25, Bucharest, Book of abstracts vol 1, p 116, 2002. ISBN 973-569-666-3
- Adriana Plesa, Gabriela Anton, Carmen C. Diaconu, M. Stoian, Rodica Repanovici – Diagnosis and / or prevention of HPV infection using viral antigens produced by gene cloning – Exhibition “Made in ROMANIA”, 2002. Bucharest
