The Comparison of Antibodies Raised Against PLRV with Two Different Approaches - Viral Particles Purification and Recombinant Production of CP

Abstract

Serology is one of the most important techniques which extensively used in different fields especially the agriculture one. Serological methods usually which are extensively used because of their specificity in disease diagnosis and relative ease of completion. The methods are widely used in plant virology include enzyme-linked immunosorbent assay, dot-blot immunoassay, immunospecific electron microscopy, and tissue-blot immunoassay. The main stone of the serology test is antiserum which is either mono or poly. Due to the high cost of the monoantisera production and its low sensitivity of the ones immunized by the purified virus particles, the recombinant protein could be good substitution. In this study, we aimed to produce polysera using the viral particles and with the recombinant coat protein of the Potato Leaf Roll Virus as well and used both in rabbit immunization separately. The sensitivity, specificity, and reactivity of the two produced sera were tested in comparing with the real-time polymerase chain reaction. Results revealed that polysera produced by recombinant coat protein are more specific and sensitive than the sera produced by the purified particles. Moreover, the results obtained by dot and tissue blot confirmed the results obtained by enzyme-linked immunosorbent assay techniques. The real time-polymerase chain reaction results were similar to that obtained by serological methods except the time and the substrate. In conclusion, production of the polysera using recombinant protein is test easy, cheap, and sensitive test for on line vial detection.