EVALUATION OF THE SENSITIVITY OF NITROCELLULOSE MEMBRANE LOW-COST ALTERNATIVES USED IN TISSUE BLOT IMMUNOASSAY (TBIA) FOR DETECTION OF SOME PLANT VIRUSES

Abstract

In an attempt of reducing the cost of tissue blot immunoassay (TBIA), some kinds of papers as alternative solid phases in place of nitrocellulose membrane (NCM), were tested for detection of three viruses: Cowpea mosaic virus (CPMV), Tomato mosaic virus (ToMV) and Zucchini yellow mosaic virus (ZYMV). Most of the examined papers developed positive reaction with all tested viruses. Canson paper (150g/m² and 300g/m²) were sensitive for detection of the tested viruses as NCM and persist in good condition during the processing. Filter paper (Whatman No. 1), local drawing paper (60g/m²), photocopy paper (80g/m²), (56g/m²) and (70g/m²) were very weak and need to taking care in handling. High glossy photo paper (200g/m²) was not effective however, it did not show any infected or healthy tissue print. Good results were obtained when both faces of NCM and Canson paper (300g/m^-2) were printed with all tested viruses. TBIA could detect CPMV in infected leaves using the unused face of NCM previously processed with 7 or 11 years ago. CPMV, ToMV and ZYMV were detected 2, 4, 6, 8 and16 days after inoculation with NCM and Canson paper (300g/m^-2). Also, the tested viruses were detected in leaf, petiole, stem and root of infected plants. ToMV could be detected using NCM and Canson paper (300g/m^-2) in naturally infected tomato plant samples.