An improved enzyme-linked immunosorbent assay (ELISA) for the determination of southern bean mosaic virus (SBMV) with linear sweep voltammetry based on a new system of p-aminophenol (PAP)-H2O2-horseradish peroxidase (...An improved enzyme-linked immunosorbent assay (ELISA) for the determination of southern bean mosaic virus (SBMV) with linear sweep voltammetry based on a new system of p-aminophenol (PAP)-H2O2-horseradish peroxidase (HRP) has firstly been developed. The enzymatic product 3-[(4-hydrox-yphenyl) amino]-4-(2-amino-5-hydroxyphenyl)-6-[ (4-hydrox-yphenyl)imino]-2,4-cyclohexadiene-l-one, produced from the oxidation of PAP with H2O2 catalyzed by HRP, yielded a sensitive linear sweep voltammetric response at - 0.45 V ( vs. SCE) in Britton-Robinson (BR) buffer solution. Based on the voltammetric peak, HRP can be measured with a detection limit of 0.4 mU/L and a linear range of 1.0-1.0 × 102 mU/ L. The detection limit for the SBMV is 8.0 ng/mL and the highest dilution ratio for the detection of infected leaf sap is 1: 1.5×105.展开更多
Imported cowpea seeds were detected with growing test,ELISA assay and RT-PCR method.The ELISA results showed that cowpea seedlings with symptoms reacted positively with antibody against Southern bean mosaic virus(SBMV...Imported cowpea seeds were detected with growing test,ELISA assay and RT-PCR method.The ELISA results showed that cowpea seedlings with symptoms reacted positively with antibody against Southern bean mosaic virus(SBMV).The 979 bp of fragment could be amplified from two positive ELISA samples using primers specific for Southern cowpea mosaic virus(SCPMV),and the sequence determination results proved that the pathogen existing in imported cowpea seeds was SCPMV.The positive ELISA results with SBMV antibody could be further confirmed by RT-PCR amplification with specific primers designed to amplify the coat protein gene and 3’ noncoding region of SCPMV and SBMV.The RT-PCR method presented here was suitable for molecular identification of SBMV and SCPMV in entry-exit plant quarantine laboratories.展开更多
基金Project supported by the National Natural Science Foundation of China (No.29775012) and the Foundation of State OPen Laboratory of Electrochemistry of Changchun Institute of Applied Chemistry.
文摘An improved enzyme-linked immunosorbent assay (ELISA) for the determination of southern bean mosaic virus (SBMV) with linear sweep voltammetry based on a new system of p-aminophenol (PAP)-H2O2-horseradish peroxidase (HRP) has firstly been developed. The enzymatic product 3-[(4-hydrox-yphenyl) amino]-4-(2-amino-5-hydroxyphenyl)-6-[ (4-hydrox-yphenyl)imino]-2,4-cyclohexadiene-l-one, produced from the oxidation of PAP with H2O2 catalyzed by HRP, yielded a sensitive linear sweep voltammetric response at - 0.45 V ( vs. SCE) in Britton-Robinson (BR) buffer solution. Based on the voltammetric peak, HRP can be measured with a detection limit of 0.4 mU/L and a linear range of 1.0-1.0 × 102 mU/ L. The detection limit for the SBMV is 8.0 ng/mL and the highest dilution ratio for the detection of infected leaf sap is 1: 1.5×105.
文摘Imported cowpea seeds were detected with growing test,ELISA assay and RT-PCR method.The ELISA results showed that cowpea seedlings with symptoms reacted positively with antibody against Southern bean mosaic virus(SBMV).The 979 bp of fragment could be amplified from two positive ELISA samples using primers specific for Southern cowpea mosaic virus(SCPMV),and the sequence determination results proved that the pathogen existing in imported cowpea seeds was SCPMV.The positive ELISA results with SBMV antibody could be further confirmed by RT-PCR amplification with specific primers designed to amplify the coat protein gene and 3’ noncoding region of SCPMV and SBMV.The RT-PCR method presented here was suitable for molecular identification of SBMV and SCPMV in entry-exit plant quarantine laboratories.
