Development of an immunochromatographic test strip for the rapid detection of fumonisin B1
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摘要: 目的:为制备一种快速检测玉米中伏马菌素B1（FB1）胶体金免疫层析试纸条。方法:采用碳化二亚胺法合成检测抗原FB1-BSA,柠檬酸三钠还原法制备胶体金溶液,辛酸-饱和硫酸铵法对抗FB1单克隆抗体腹水进行纯化,将金标抗体喷于金标垫,检测抗原FB1-BSA（T线）和羊抗鼠二抗（C线）喷涂于硝酸纤维素膜（NC膜）。结果:得到的单克隆抗体效价为1.28×105。该试纸条的NC膜喷涂的检测抗原浓度为200μg/m L,羊抗鼠二抗浓度为1.0 mg/mL,喷涂量分别为0.74μL/cm,试纸条灵敏度为20 ng/mL,检测时间只需5 min,试纸条于4℃至少可保存12个月。结论:采用制备的试纸条对玉米实际样品进行检测,检测结果与高效液相和酶联免疫吸附法检测结果相一致,说明该试纸条适合现场快速检测伏马菌素B1。Abstract: Objective : An immunochromatographic test strip( ICG) had been developed for rapid detection of fumonisin B1 residues in maize samples. Methods:For this purpose,FB1 coupled to bovine serum albumin(BSA) via the modified EDCA method was prepared as capture antigen. The colloidal gold was prepared by the reduction of tetrachloroauric(III) acid trihydrate with citric acid trisodium salt. Results :Using an antibody purified by the ammonium sulfate precipitation,and a titer of the antibody was about 1.28 ×105by ELISA. The ICG was composed of NC membrane,sample pad,probe pad(colloidal gold-monoclonal antibody probes for FB1 and absorbent pad. Moreover,the sensitivity,specificity and stability of the ICG were also detected. A total of 0.74 μL/cm of FB1-BSA(200 μg/m L) and the goat anti-mouse Ig G antibody(1.0 mg/m L) were sprayed onto the NC membrane as the test(T line) and control lines(C line),respectively. The sensitivity of the test strip was20 ng/m L. The test could be accomplished within 5 min. The stability of the ICG was about 12 months at 4℃.Conclusion:Analysis of FB1 in maize samples revealed that data obtained from the ICG were in good agreement with those obtained from HPLC and ELISA. The results demonstrated that the ICG could be used as qualitative tool for rapid screening of FB1on-site.
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Keywords:
- rapid detection /
- immunochromatographic strip test /
- fumonisin B1 /
- maize
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