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Development of Rapid Immune-gold Strip Kit for On-Site Diagnosis of Tomato spotted wilt virus |
Ju-Yeon Yoon1, Gug-Seoun Choi2, In-Sook Cho2 and Seung-Kook Choi2* |
1Department of Horticulture, Biotechnology and Landscape, Seoul Women’s University, Seoul 139-774, Korea 2Virology Unit, Department of Horticultural Environment, National Institute of Horticultural and Herbal Science, Rural Development Administration, Suwon 441-440, Korea |
Correspondence:
<p>Seung-Kook Choi</p><p>Tel : +, Seung-Kook Choi Tel : +82-31-290-6236 Fax: +82-31-290-6259 E-mail: viroid73@gmail.com |
Received: September 24, 2013 Revised: January 27, 2014 Accepted: March 04, 2014 |
Abstract |
A rapid, user-friendly and simple immune-chromatographic dipstick kit named ‘rapid immune-gold strip’ (RIGS) kit was developed in a novel single strip format to detect on-site detection of Tomato spotted wilt virus (TSWV). Immunoglobulin G (IgG) from polyclonal antisera raised in rabbits against TSWV was purified through protein- A affinity chromatography and then the purified TSWV-IgG was conjugated to colloidal gold nano-particles which served as a test line on nitrocellulose membrane. Protein A that non-specifically binds to TSWV antibody was used as a control line on the same strip. The diagnosis process with the TSWV-RIGS 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 2-5 minutes. The flow of the complexes of gold particles coated with TSWV-IgG and a crude sap from TSWV-infected pepper, tobacco and tomato plants resulted in intensive color formed on the test lines proportional to the concentrations of TSWV. The RIGS-TSWV kit did not show any cross-reactions against other tomato-infecting viruses unrelated to TSWV. These results indicate that the TSWV-RIGS kit is highly sensitive and is not required for laboratory training and experience prior to testing. The TSWV-RIGS kit is suitable for on-site detection of suspect TSWV-infected plants as well as for laboratory diagnosis. |
Key Words:
Detection, Diagnosis, IgG, Rapid Immune-Gold Strip, On-site, Tomato spotted wilt virus |
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