Monoclonal Antibodies against the Recombinant Nucleocapsid Protein of Tomato spotted wilt virus and its Application in Virus Detection

Tomato spotted wilt virus (TSWV) is the type member of the tospovirus genus and causes significant losses in a wide range of economically important ornamental and vegetable crops worldwide. The nucleocapsid gene, located on the ambisense S RNA segment of TSWV was expressed in Escherichia coli using pET‐32a as vector and correct expression of recombinant protein was confirmed by Western blot using an anti‐TSWV monoclonal antibody (MAb). The recombinant protein was purified using Ni‐NTA agarose and the purified protein was used for the production of MAbs. Three murine MAbs against the recombinant nucleocapsid protein were produced. Triple antibody sandwich enzyme‐linked immunosorbent assay and immunocapture RT‐PCR methods were then established for reliable and efficient detection of TSWV using the produced MAbs.