The sequence encoding an E2 main antigen glycoprotein of the C strain of classical swine fever virus (CSFV) was highly expressed in the host cell E. coli BL21–CodonPlus (DE3)–RIL using the pGEX-4T-1 expression vecto...The sequence encoding an E2 main antigen glycoprotein of the C strain of classical swine fever virus (CSFV) was highly expressed in the host cell E. coli BL21–CodonPlus (DE3)–RIL using the pGEX-4T-1 expression vector and the soluble recombinant product was purified with Glutathione Sepharose TM4B by centrifugation. The soluble recombinant protein showed good immune reactions and was confirmed by Western blot using anti-CSFV-specific antibodies. Then an indirect ELISA with the purified E2 protein as the coating antigen was established to detect antibody against CSFV. The result revealed that the optimal concentration of coated antigen was 0.6 μg/well and the optimal dilution of serum was 1:80. The positive cut-off value of this ELISA assay was OD tested serum / OD negative serum≥2.1. The E2-ELISA method was evaluated by comparison with the indirect hemagglutination test (IHAT). When a total of 100 field serum samples were tested the sensitivity and specificity were 90.3% and 94.7% respectively. Specificity analysis showed that there were no cross-reactions between BVD serum and the purified E2 protein in the E2-ELISA.展开更多
Extraction of AchE, relationship between substrate and enzyme concentration, and inhibition effects of the agrochemicals to AchE are discussed in this paper. Through the re-search, the proper AchE concentration for hy...Extraction of AchE, relationship between substrate and enzyme concentration, and inhibition effects of the agrochemicals to AchE are discussed in this paper. Through the re-search, the proper AchE concentration for hydrolysis of 1 ml 1mmol/L substrate and I50 val-ues of the agrochemicals to AchE are decided. It is proved that Asch-DTNB method is a rapid test tool for agrochemical residues in fruits and vegetables.A rapid test card has been developed with sensitivity of 0.05mg/L.展开更多
In this study, the susceptibility of three populations of cotton aphid, Aph＆ gossypii Glover （Hemiptera：Aphididae） was assayed to imidacloprid （35SC） and thiametoxam （50WG）. The involvement of metabolic enzyme...In this study, the susceptibility of three populations of cotton aphid, Aph＆ gossypii Glover （Hemiptera：Aphididae） was assayed to imidacloprid （35SC） and thiametoxam （50WG）. The involvement of metabolic enzymes in the resistance strain of cotton aphid to the neonicotinoids was determined by the biochemical biomarkers and the resistance mechanism was determined as CaE. In another study, three different bioassay experiments were designed for detecting the susceptibility of cotton aphid to imidacloprid and thiametoxam and the effect of these two insecticides on the enzymatic activity of cotton aphid was assessed in the adult aphids treated with three different bioassay methods using a modified version of the FAO dip test, residue bioassay procedure and starvation method. Our findings suggested that the type of bioassay methods is very important when aphids＇ populations assess for the resistance against the neonicotinod insecticides. It has shown the starvation method is the most reliable method compared with other methods.展开更多
基金Society Commonweal Study of China (2001DIA10006)
文摘The sequence encoding an E2 main antigen glycoprotein of the C strain of classical swine fever virus (CSFV) was highly expressed in the host cell E. coli BL21–CodonPlus (DE3)–RIL using the pGEX-4T-1 expression vector and the soluble recombinant product was purified with Glutathione Sepharose TM4B by centrifugation. The soluble recombinant protein showed good immune reactions and was confirmed by Western blot using anti-CSFV-specific antibodies. Then an indirect ELISA with the purified E2 protein as the coating antigen was established to detect antibody against CSFV. The result revealed that the optimal concentration of coated antigen was 0.6 μg/well and the optimal dilution of serum was 1:80. The positive cut-off value of this ELISA assay was OD tested serum / OD negative serum≥2.1. The E2-ELISA method was evaluated by comparison with the indirect hemagglutination test (IHAT). When a total of 100 field serum samples were tested the sensitivity and specificity were 90.3% and 94.7% respectively. Specificity analysis showed that there were no cross-reactions between BVD serum and the purified E2 protein in the E2-ELISA.
文摘Extraction of AchE, relationship between substrate and enzyme concentration, and inhibition effects of the agrochemicals to AchE are discussed in this paper. Through the re-search, the proper AchE concentration for hydrolysis of 1 ml 1mmol/L substrate and I50 val-ues of the agrochemicals to AchE are decided. It is proved that Asch-DTNB method is a rapid test tool for agrochemical residues in fruits and vegetables.A rapid test card has been developed with sensitivity of 0.05mg/L.
文摘In this study, the susceptibility of three populations of cotton aphid, Aph＆ gossypii Glover （Hemiptera：Aphididae） was assayed to imidacloprid （35SC） and thiametoxam （50WG）. The involvement of metabolic enzymes in the resistance strain of cotton aphid to the neonicotinoids was determined by the biochemical biomarkers and the resistance mechanism was determined as CaE. In another study, three different bioassay experiments were designed for detecting the susceptibility of cotton aphid to imidacloprid and thiametoxam and the effect of these two insecticides on the enzymatic activity of cotton aphid was assessed in the adult aphids treated with three different bioassay methods using a modified version of the FAO dip test, residue bioassay procedure and starvation method. Our findings suggested that the type of bioassay methods is very important when aphids＇ populations assess for the resistance against the neonicotinod insecticides. It has shown the starvation method is the most reliable method compared with other methods.
