토마토덤불위축바이러스 현장진단용 급속 면역 금 나노입자 막대 키트 개발

Abstract

A rapid, user-friendly and simple immune-chromatographic dipstick kit named ‘rapid immuno-gold strip’ (RIGS) kit was developed in a novel single strip format to detect on-site detection of Tomato bushy stunt virus (TBSV). Immunoglobulin G (IgG) from polyclonal antisera raised in rabbits against TBSV was purified through protein-A affinity chromatography and then the purified TBSV-IgG was conjugated to colloidal gold nano-particles which served as a test line on nitrocellulose membrane. Protein A that non-specifically binds to TBSV antibody was used as a control line on the same strip. The diagnosis process with the RIGS-TBSV involves simply grinding the suspect plant sample in a bag that contains the extraction buffer and inserting the strip the bag. Results can be seen in 5-10 minutes. The flow of the complexes of gold particles coated with TBSV-IgG and a crude sap from TBSV infected tomato plants resulted in intensive color formed on the test lines proportional to the concentrations of TBSV. The RIGS-TBSV kit did not show any cross-reactions against Tomato spotted wilt virus and Tomato yellow leaf curl virus that are major causal viruses. These results indicate that the RIGS-TBSV kit is highly sensitive and is not required for laboratory training and experience prior to testing. The RIGS-TBSV kit is suitable for on-site determination of suspect TBSV-infected plants as well as for laboratory diagnosis.