Interaction of Plum Pox Virus with Specific Colloidal Gold-Labeled Antibodies and Development of Immunochromatographic Assay of the Virus

• Published in: Biochemistry (Moscow) Vol. 75; no. 11; pp. 1393 - 1403
• Main Authors: Byzova, N. A, Safenkova, I. V, Chirkov, S. N, Avdienko, V. G, Guseva, A. N, Mitrofanova, I. V, Zherdev, A. V, Dzantiev, B. B, Atabekov, J. G
• Format: Journal Article
• Language: English
• Published: Dordrecht Dordrecht : SP MAIK Nauka/Interperiodica 01.11.2010; SP MAIK Nauka/Interperiodica; Springer; Springer Nature B.V
• Subjects: Animals; Antibodies, Monoclonal; Atomic force microscopy; Biochemistry; Biomedical and Life Sciences; Biomedicine; Bioorganic Chemistry; Chromatography, Thin Layer; Colorimetry; Enzyme-Linked Immunosorbent Assay; Flocculation; Gold Colloid; Health aspects; Immune system; Immunohistochemistry; Life Sciences; Limit of Detection; Mice; Mice, Inbred BALB C; Microbiology; Monoclonal antibodies; Nanoparticles; Particle Size; Pay-per-view television; Plant diseases; Plant Diseases - virology; Plant Leaves - virology; Plum pox virus; Plum Pox Virus - immunology; Prunus; Prunus - virology; Reagent Strips; Virology - methods; Viruses; colloidal gold; transmission electron microscopy; atomic force microscopy; immunochromatographic assay; plum pox virus; monoclonal antibodies
• ISSN: 0006-2979; 1608-3040
• DOI: 10.1134/S000629791011012X

Two monoclonal antibodies (mABs) raised against plum pox virus (PPV) were shown to recognize its D, M, and C strains. Conjugates of the antibodies with colloidal gold (CG) nanoparticles averaging 26 nm in diameter were synthesized. The binding constants of PPV with both the native and conjugated mABs were determined using a Biacore X device. The complexes between the CG-mAB conjugates and plum pox virions were examined by means of transmission electron and atomic force microscopy. Using the conjugates with optimal component ratio, an express immunochromatographic assay of PPV was developed with a detection limit of 3 ng/ml and duration of 10 min. The assay was tested for PPV detection in sam- ples of stone fruit tree leaves and demonstrated a good compatibility with the data obtained by “sandwich”-ELISA. The developed assay can be used in the field and applied for monitoring viral infection and for quarantine purposes.