Immunodetection of Furcraea Necrotic Streak Virus-FSNV in Fique Plants (Furcraea Macrophylla Baker) Using A Polyclonal Antibody IgY Produced in Chicken Egg Yolk
The necrotic streak of the fique (Furcraea spp.) or “Macana” disease is considered the most limiting disease for this crop in Colombia, whose causal agent is the Furcraea Necrotic Streak Virus - FSNV (RNA +). Currently, there are no strategies to control the disease, being necessary to develop methods for early detection of this disease in the planting material before being taken to the field. In this study, polyclonal antibodies produced in egg yolk (IgY) were assesses for detection FSNV. Two immunoenzymatic methodologies were standardized: Dot Blot Immunobinding Assay (DBIA) and Enzyme Linked Immunosorbent Assay (ELISA), determining their specificity and sensitivity. The detection limit by DBIA corresponded to 8 µg/mL of purified virus suspension using 10 µg/mL of primary antibody. In the ELISA test, the primary antibody concentration of 3 µg/mL (1:800 dilution) detected the antigen at concentrations between 10 and 70 µg/mL. The polyclonal antibody anti-FNSV IgY allowed the detection of FNSV in samples of purified virus and extracts of roots and leaves of fique plants with symptoms of “Macana” disease and did not produce any signal with the control samples. Results showed the potential of using egg yolk IgY for immunological test as a novel approach for the early detection of FNSV in fique plants.
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