Establishment of ELISA method for testing tobacco mosaic virus (TMV) infecting Zingiber officinale Rosc

VernacularTitle:检测侵染生姜的烟草花叶病毒ELISA的建立
Author: Chuanbao WEI ; Hui DENG
Keywords: Tobacco mosaic virus(TMV); Zingiber offcinale Rosc.; ELISA method
From: Chinese Journal of Immunology 2017;33(9):1355-1358,1360
CountryChina
Language:Chinese
Abstract: Objective:In order to establish ELISA method for testing tobacco mosaic virus (TMV) infecting Zingiber officinale Rosc.Methods: Tobacco mosaic virus(TMV) infecting Zingiber officinale Rosc was tested by RT-PCR.Zingiber officinale Rosc leaves which only contained TMV were choiced.TMV particle was purified by centrifugation method.TMV CP was purified through preparation electrophoresis including 12％ SDS-PAGE first and then 5％-20％ gradient SDS-PAGE.Polyacrylamide gel contained TMV CP was ground into suspension.Mice were immuned with the suspension and antiserum was obtained.Antiserum quality was tested by Western blot and ELISA test.IgG was purified through affinity chromatography method.IgG solution was concentrated and dialyzed to a suitable concentration.The IgG then mixed with glycerol.Results: IgG in antiserum only combined with TMV CP protein and it could combine with nature TMV particle CP protein.Its quality was up to standard.Conclusion: Establishment of ELISA method for testing TMV infecting Zingiber offcinale Rosc is successfull by using this IgG.