Pemanfaatan Antibodi Monoklonal dalam Immunoassay untuk Deteksi Baculovirus oryctes

https://doi.org/10.22146/jpti.12384

Susamto Somowiyarjo(1*), Hery Haryanto(2), Sri Wening(3)

(1) Fakultas Pertanian Universitas Gadjah Mada
(2) Fakultas Pertanian Universitas Mataram
(3) Fakultas Pertanian Universitas Gadjah Mada
(*) Corresponding Author

Abstract


The application of non-precoated Indirect Enzyme-Linked Immunosorbent Assay (ELISA) and Dot Immunobinding Assay (DIBA) employing monoclonal antibodies (MCA) against Yogyakarta isolate of Baculovirus oryctes Huger. was described. The MCA-Bv-4 having subclass of IgG2a and titer in vitro of 10^4 - 10^5 proved to be useful antibody for virus detection. The great potential of I-ELISA using MCA-Bv-4 has been it's specificity being able to discriminate between healthy an virus-infected coconut beetle (Oryctes rhinoceros L.). The assay could also differentiate between B. oryctes and Monodon baculovirus, the pathogen of shrimp disease. The best tissue for preparing virus antigen from crude extract was found to be the midgut of the beetle. Head, thorax, abdomen and tibia were not suitable for preparing the test antigen. The crude extract of beetle showed high endogenous enzyme activity to the substrate of DIBA, which precluded the detection of B. oryctes using DIBA. The MCA-Bv-4 could be used to improve the monitoring of the virus to support the program of biological control of coconut beetle using B. oryctes.


Keywords


monoclonal antibodies; immunoassay; Baculovirus oryctes

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References

Banttari, E.E. & P.H. Goodwin. 1985. Detection of Potato Viruses S, X, and Y by Enzyme-Linked Immunosorbent assay on Nitrocellulose Membranes (Dot-ELISA). Plant Dis. 69: 202-205.

Crook. E.N. & C.C. Payne. 1980. Comparison of three Methods of ELISA for Baculoviruses. J. gen. Virol. 46: 29-37.

Galfre, G., S.C. Howe, C. Milstein G.W. Butcer &. J.C. Boward. 1977. Antibodies to Major Histocompatibility Antigens Produced by Hybrid Cell Lines. Nature 266: 550-552.

Hibi, T. & Y. Saito. 1985. A Dot Immunobinding Assay for the Detection of Tobacco Mosaic Virus in Infected Tissues. J. gen. Virol. 66: 1191-1194.

Kalshoven, L.G.E. I 98 I. The Pest of Crops in Indonesia. P.T. lchtiar Baru-Van Hoeve, Jakarta. 701 p.

Kobayashi, J. & S. Somowiyarjo. 1995. Properties of Baculovirus oryctes Isolated in Indonesia. Indon. J. Plant Prat, I: 41-45.

Kohler, G. & C. Milstein. 1975. Continous Culture of Fused Cells Secreting Antibodies of Predefined Specifity. Nature 256: 495-497.

Koenig, R. 1981. Indirect-ELISA Methods for the Broad Specifity Detection of Plant Viruses. J. gen. Virol. 67: 53-62.

Mangoendihardjo. S. 1995. Pengendalian Hayati Komponen Utama Pengelolaan Jasad Pengganggu. Pidato Pengukuhan Jabatan guru Besar dalam Ilmu Hama Tumbuhan pada Fakultas Pertanian tanggal 22 Maret 1995, Yogyakarta. 26 hlm.

Mink, G.I, W.E. Howell & P.R. Fridlund. 1985. Apple tip Leaf Antigens That Cause Spurious Reactions with Tomato Ringspot Virus Antisera in Enzyme-Linked Immunosorbent Assay. Phvtopathologv 75: 325-329.

Murwantoko. 1997. Isolasi dan Karakterisasi Parsial Biomolekul Beberapa Virus Penyebab Penyakit Udang. Tesis S2 UGM, Yogyakarta (Tidak dipublikasikan). 64 hlm.

O'Reilly, D.R., L.K. Miller & V.A. Luckow. 1992. Baculovirus Expression Vectors, A Laboratory Manual. W.H. Freeman and Company. New York. 347 p.

Schreier. M., G. Kohler, H. Hengartner, C. Berek, M. Trucco, L. Forni, T. Staehelin, J. Stocker & B. Takaes. 1980. Hybridoma Techniques. Cold Spring Harbor, New York. 65 p.

Sikora. K. & H.M. Smedley. 1984. Monoclonal Antibodies. Blackwell Scientific Publications. London. 132 p.

Soemarwoto, O. 1996. Pembangunan Pertanian Berkelanjutan. Orasi llmiah pada Peringatan Setengah Abad Fakultas Pertanian UGM tanggal 27 September 1996. 18 hlm.

Somowiyarjo. S., N. Sako & F. Nonaka. 1985. Application of Non-Precoated Indirect ELISA for Detecting Zucchini Yellow Mosaic Virus. Ann. Phytopath. Soc. Japan 51: 569-575.

Somowiyarjo, S., N. Sako & F. Nonaka. 1986. Comparison of Two Indirect ELISA Procedures for Detecting Zucchini Ye1low Mosaic Virus. Ann. Phytopath. Soc. Japan 52: 653-659.

Somowiyarjo. S., N. Sako & F. Nonaka. 1988. The Use of Monoclonal Antibody for Detecting Zucchini Yellow Mosaic Virus. Ann. Phytopath. Soc. Japan 54: 436-433.

Somowiyarjo, S., Y.B. Surnardiyono, S. Hartono, Triharso & J. Kobayashi. 1995. Propagation and Purification of Baculovirus oryctes Huger. Im/on. J. Plant. Prot. I: 38-40.

Sumardiyono, Y.B. 1995. Elektron Mikroskopi dan lmunogenisitas Baculovirus lsolat Yogyakarta. lndon. J. Plant Prot. I: 46-49.

Summers, M.D. 1977. Baculoviruses (Baculoviridae), p. 3-9. In Maramorosch. K. (ed.) The Atlas of Insect and Plant iruses.. Acad. Press, New York.



DOI: https://doi.org/10.22146/jpti.12384

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