Palavras-chave
Dengue
MAC-ELISA
Immunoblotting
proteínas recombinantes
MAC-ELISA
Immunoblotting
proteínas recombinantes
Como Citar
1.
Luisa Barbosa M, Maria M. Souza-Felippe J, A. Fonseca B, N. Manson P, I. B. Ferreira I, Antonio Jerez J. Detecção de vírus Dengue sorotipos 1 e 2 pela técnica de immunoblotting utilizando proteínas recombinantes. Rev Inst Adolfo Lutz [Internet]. 30º de dezembro de 1999 [citado 29º de abril de 2024];58(2):121-7. Disponível em: https://periodicos.saude.sp.gov.br/RIAL/article/view/36704
Resumo
Os vírus dengue são patógenos que vêm afetando milhões de pessoas durante os dois últimos séculos. No presente trabalho, comparou-se a técnica tradicional de MAC-ELISA e o teste de Immunoblotting, utilizando-se proteínas recombinantes, para pesquisa de anticorpos da classe M, contra os vírus Dengue sorotipos 1 e 2. Foram testadas 100 amostras de soro de pacientes, com suspeita clínica de dengue, por MAC-ELISA e Immunoblotting. Estes soros quando submetidos ao MAC-ELISA resultaram 56 positivos para dengue e 44 negativos. Estas mesmas amostras avaliadas por Immunoblotting revelaram 56 soros negativos, 36 soros positivos para DEN-l, 6 para DEN-2 e 2 para DEN-l e 2. Proteínas recombinantes (DEN-l e DEN-2) utilizadas como antígeno em Immunoblotting possibilitam um diagnóstico diferencial dos sorotipos de dengue circulantes em determinada área, enquanto o teste de MAC-ELISA apesar de sensível e rápido não tipifica o sorotipo de dengue, dado importante durante os períodos de epidemias.
Referências
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21. RICE, e.M.; STRAUSS, E.G. & STRAUSS, J.H. Structure of the flavivirus genome. ln: S. Schlesinger & M.J. Schlesinger. Togaviridae and Flaviviridae, New York: Plenum, 1986, 279326.
22. ROEHRIG, J.T.; JOHNSON, A.J.; HUNT, A.R; BOLIN, RA. & CHU, M.C.. Antibodies to dengue 2 virus E-glycoprotein synthetic peptides identify antigenic conformation. Virology, 177: 668-675, 1990.
23. ROEHRIG, J.T.; MATHEWS, J.H. & TRENT, D.W .. ldentification of epitopes on the E glycoprotein of St. Louis encephalitis virus using monoclonal antibodies. Virology, 128: 118, 1983.
24. SAMBROOK, I; FRITSCH, E.F. & MANIA TIS , T. Moleeular cloning: a laboratory manual. 2nd. ed., New York, Cold Spring Harbor Laboratory Press, 1989.
25. SELB, B. & WEBER, B.. A study 01'human reovirus IgG and IgA antibodies by ELISA and Western blot. J. ViroI. Methods, 47: 15-26, 1994.
26. SMITH, G.W. & WHIGHT, P.L Synthesis 01'proteins and glycoproteins in dengue type 2 virusinfected Vero and Aedes albopictus cells. 1. Gen. Virol., 66: 559-571, 1985.
27. TOWBIN, H.; STAEHELIN, T. & GORDON, L Eletrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proe. Natl. Aead. Sei. USA, 76: 4350-4354, 1979.
28. VOLLER, A. &BIDWELL, D.. Enzime-linked immunosorbent assay. ln Rose ,N.R.I Fridman, H & Fahey, IH. eds. Manual of Clinical Laboratory Immunology. Washington, De American Society for Microbiology. 99-109, 1986
29. WESTAWAY, E.O. Flavivirus rep1ication strategy. Adv. Vírus Res., 33: 45-90 1987.
30. WESTAWAY, E.O. Rep1ication of Flavivirus ln: The Togaviruses: Biology, Structure, Replication. New York: Academic Press, 1980,531- 581.
2. AUSUBEL, EM.; BRENT, R; KINGSTON, RE.; MOORE, D.D.; SEIDMAN,J.G. & STRURHL, RE. Current protocols in molecular hiology. New York, John Wiley & Sons, 1989.
3. BURNETTE, W.N.. "Western blotting": eletropho-retic transfer of proteins from sodium dodecyl sulfate-polyacrylarnide gels to unrnodified nitrocellulose and radiographic detection with antibody and radiated protein A Anal. Biochem., 112: 195-203, 1981.
4. CHAMBERS, T.J., HAHN, C.S., GALLER, R & RICE, C.M., Flavivirus genome organization expression, and replication. Annu. Rev. Microhiol., 44: 649- 688, 1990.
5. CHURDBOONCHART, v, BHAMARAPRAVITI, N.; PEAMPRAMPRECHA, S. & SIRINAVIN, S.. Antibodies Against Dengue viral proteins in primary and Secondary Dengue Hemorrhagic Fever. Am. J. Trop. Med. Hyg., 44: 481-493,1991.
6. CHURDBOONCHART, v, BHAMARAPRAVITI, N.; PEAMPRAMPRECHA, S. PINTHONG, C.; SIRINAVIN, S. & CHAIYOTHA, A. Antibodies to dengue viral polypeptides. r. Sensitivity and specificity of the viral-antigen-stripsl enzyme immunoassay. South. Asian J. Trop. Med. Publie Health, 18: 362-372, 1987.
7. CLARKE, D.H. & CASALS, L Tecniques for hemagglutination and hemagglutination- inhibition with arthropod-borne viruses. Am. J. Trop. Med. Hyg., 7: 551, 1958.
8. FONSECA B.A.L., KHOSHNOOD K.,SHOPE RE. & MANSON P.W.. Flavivirus Type- Specific Antigens Produced from Fusions of a Portion of the E Protein Gene with the Escherichia coli TrpE Gene. Am. J. Trop. Med. Hyg., 44: 500-508, 1991.
9. GUBLER, D.L DenguelDengue Hemorrhagic fever in the Americas: Prospects for the year 2000. pg-19-27. ln: Dengue: A Wordwide Problem, a Common Strategy. Ed. by S.B. Halstead & Gomez- Dantes, 1992.
10. HALL, R.A.; KAY,H.B.; BURGESS, G.W.; CLANCY, P.& FINNING, 1.D.. Epitope analysis of the envelope and non-structural glycoproteins of Murray Valley encephalitis vírus. J. Gen. Virol., 71 :2923-2930, 1990.
11. HALSTEAD, S.B .. Pathogenesis of Dengue: Challenges to Molecular Biology- Seienee, 239: 476- 481, 1988
12. HASEGAWA, W.; YOSHIDA, M.; SHIOSAKA, T.; FUJITA, S. & KOBAYASHI, Y.. Mutations in the envelope protein of Japanese encephalitis virus affect entry into cultured cells and virulence in mice. Virology, 191: 158-165, 1992.
13. HEINZ, F.X .. Epitope mapping of flavivirus glycoproteins. Adv. Virus Res., 31: 103-168, 1986.
14. HENCHAL, E.A.; McCOWN, J.M.; BURKE, D.S.; SEGUIN, M.e. & BRANDT, W.E .. Epitopic analysis of antigenic determinants on the surface of dengue-2 virions using monoclonal antibodies. Am. J. Trop. Med. Hyg., 34: 162-167, 1985.
15. KUNO, G.; GÓMEZ, L & GUBLER, D.L Detecting artificial anti-dengue IgM Immune complexes using an enzyme-linked immunosorbent assay. Am Trop. Med. Hyg., 36: 153-159, 1987.
16. LINDO, J.F.; CONWAY,D.J.; ATKINS, N.S.; BIANCO,A.E.; ROBINSON, RD. & BUNDY, D.A.P .. Prospective evaluation of enzyme-linked immunosorvent assay and immunoblot methods for the diagnosis of endemic Trongyloides stercoralis infection. Am J. Trop. Med. Hyg., 51: 175-179, 1994.
17. MANDL, C.W.; HEINZ, F.X.; STOCKL, E. & KUNZ, c.. Genome sequence of tick-borne encephalitis virus ( Western subtype) and comparative analysis of nonstructural proteins with other Flaviviruses. Virology, 173: 291-301, 1989.
18. MASON, P.W.; DALRYMPLE, J.M.; GENTRY, M.K.; McCOWN, IM.; HOKE, c.n., BURKE, D.S; FOURNIER, M.J. & MASON, T.L.. Molecular Characterization of the Japanese Encephalitis Virus Structural Glycoprotein. J. Gen. Virol., 70: 2037- 2049, 1989.
19. MASON, P.W.; ZUGEL, M.U.; SEMPRONI, A.R; FOURNIER, M.J. & MASON, T.L.. The antigenic structure of dengue type 1 virus envelope and NS 1 proteins expressed in Escherichia coli. J. Gen. Virol., 71: 2107-2114, 1990.
20. PAN AMERICAN HEALTH ORGANIZATION (PAHO). Dengue and dengue hemorrhagic fever in the Americas: Guidelines for prevention and control. Washington, o.c. PAHO, 1994.
21. RICE, e.M.; STRAUSS, E.G. & STRAUSS, J.H. Structure of the flavivirus genome. ln: S. Schlesinger & M.J. Schlesinger. Togaviridae and Flaviviridae, New York: Plenum, 1986, 279326.
22. ROEHRIG, J.T.; JOHNSON, A.J.; HUNT, A.R; BOLIN, RA. & CHU, M.C.. Antibodies to dengue 2 virus E-glycoprotein synthetic peptides identify antigenic conformation. Virology, 177: 668-675, 1990.
23. ROEHRIG, J.T.; MATHEWS, J.H. & TRENT, D.W .. ldentification of epitopes on the E glycoprotein of St. Louis encephalitis virus using monoclonal antibodies. Virology, 128: 118, 1983.
24. SAMBROOK, I; FRITSCH, E.F. & MANIA TIS , T. Moleeular cloning: a laboratory manual. 2nd. ed., New York, Cold Spring Harbor Laboratory Press, 1989.
25. SELB, B. & WEBER, B.. A study 01'human reovirus IgG and IgA antibodies by ELISA and Western blot. J. ViroI. Methods, 47: 15-26, 1994.
26. SMITH, G.W. & WHIGHT, P.L Synthesis 01'proteins and glycoproteins in dengue type 2 virusinfected Vero and Aedes albopictus cells. 1. Gen. Virol., 66: 559-571, 1985.
27. TOWBIN, H.; STAEHELIN, T. & GORDON, L Eletrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proe. Natl. Aead. Sei. USA, 76: 4350-4354, 1979.
28. VOLLER, A. &BIDWELL, D.. Enzime-linked immunosorbent assay. ln Rose ,N.R.I Fridman, H & Fahey, IH. eds. Manual of Clinical Laboratory Immunology. Washington, De American Society for Microbiology. 99-109, 1986
29. WESTAWAY, E.O. Flavivirus rep1ication strategy. Adv. Vírus Res., 33: 45-90 1987.
30. WESTAWAY, E.O. Rep1ication of Flavivirus ln: The Togaviruses: Biology, Structure, Replication. New York: Academic Press, 1980,531- 581.
Este trabalho está licenciado sob uma licença Creative Commons Attribution 4.0 International License.
Copyright (c) 1999 Maria Luisa Barbosa, Júlia Maria M. Souza-Felippe, Benedito A. Fonseca, Peter N. Manson, Ivani I. B. Ferreira, José Antonio Jerez
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