Development and Evaluation of a Novel ELISA for Detection of Antibodies against HTLV-I Using Chimeric Peptides
AbstractWe aimed to develope a peptide-based indirect ELISA to detect antibodies against Human T-lymphotropic virus type I (HTLV-I). Two chimeric peptides (CP-1 and CP-2) were designed using linear immunodominant epitopes of gp-46-I, and gp21-I proteins, according to the sequence from Uniprot database. These peptides were studied initially in the ELISA using infected sera. The most promising peptideCP-1, was used to develop a peptide ELISA for detection of HTLV-I infected sera. The optimal conditions for CP-1ELISA were: the optimum coating buffer was 100mM NaHCO3, pH 9.6; coating peptide concentration was 10 µg/mL; the optimal blocking buffer was5% fetal bovine serum (FBS); the secondary antibody concentration was 1:2000; and serum dilution was 1:20. 20serum samples from HTLV-I infected patients were evaluated by ELISA developed. CP-1 showed high antigenicity while lacking any cross-reactivity with normal human sera. The results of evaluations indicated that in comparison with commercial ELISA, CP-1 ELISA showed good sensitivity and specificity. With further validation, CP-1as described in the present study could be introduced as novel reliable and cost-effective candidates for the high-specific screening of HTLV-I/-II infections in endemic regions.
1. Ishitsuka K, Tamura K. Human T-cell leukaemia virus type I and adult T-cell leukaemia-lymphoma. Lancet Oncol 2014;15(11):e517-26.2. Montesdeoca Andrade MJ, Correa Diaz EP, Buestan ME. HTLV-1-associated myelopathy in a solid organ transplant recipient. BMJ Case Rep 2016;2016.3. Iwanaga M, Watanabe T, Utsunomiya A, Okayama A, Uchimaru K, Koh KR, et al. Human T-cell leukemia virus type I (HTLV-1) proviral load and disease progression in asymptomatic HTLV-1 carriers: a nationwide prospective study in Japan. Blood 2010;116(8):1211-9.4. Marano G, Vaglio S, Pupella S, Facco G, Catalano L, Piccinini V, et al. Human T-lymphotropic virus and transfusion safety: does one size fit all? Transfusion 2016;56(1):249-60.5. Thorstensson R, Albert J, Andersson S. Strategies for diagnosis of HTLV-I and -II. Transfusion 2002;42(6):780-91.6. Heydari Zarnagh H, Hassanpour K, Rasaee MJ. Constructing Chimeric Antigen for Precise Screening of HTLV-I Infection. Iran J Allergy Asthma Immunol 2015;14(4):427-36.7. Cossen C, Hagens S, Fukuchi R, Forghani B, Gallo D, Ascher M. Comparison of six commercial human T-cell lymphotropic virus type I (HTLV-I) enzyme immunoassay kits for detection of antibody to HTLV-I and -II. J Clin Microbiol 1992;30(3):724-5.8. Hernandez Marin M, Marquez Bocalandro Y, Vallejo RV, et al. Use of a chimeric synthetic peptide from the core p19 protein and the envelope gp46 glycoprotein in the immunodiagnosis of HTLV-II virus infection. Prep Biochem Biotechnol 2003;33(1):29-38.9. Li J, Zhang WB, McManus DP. Recombinant antigens for immunodiagnosis of cystic echinococcosis. Biol Proced Online 2004;6:67-77.10. Marin MH, Rodriguez-Tanty C, Higginson-Clarke D, Bocalandro YM, Pena LP. Study of the peptide length and amino acid specific substitution in the antigenic activity of the chimeric synthetic peptides, containing the p19 core and gp46 envelope proteins of the HTLV-I virus. Biochem Biophys Res Commun 2005;336(3):983-6.11. Yamada K, Kuroda N, Washitani Y, Shiraki H, Maeda Y. Enzyme immunoassay with synthetic peptides to detectanti-HTLV-I antibodies. Clin Chem 1992;38(5):699-703.12. Hernandez M, Selles ME, Pozo Pena L, Gomez I, Melchor A. Antigenicity of chimeric synthetic peptides based on HTLV-1 antigens and the impact of epitope orientation. Biochem Biophys Res Commun 2000;276(3):1085-8.13. Heydari Zarnagh H, Ravanshad M, Pourfatollah AA, Rasaee MJ. Expression and Purification of a Novel Computationally Designed Antigen for Simultaneously Detection of HTLV-1 and HBV Antibodies. Iran J Allergy Asthma Immunol 2015;14(2):168-78.