Zhou Chennan, Zhang Xueyin, Huang Xinxin, Guo Xishan, Cai Qiang, Zhu Songming
College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou 310058, China.
Shanghai Entry-Exit Inspection and Quarantine Bureau, Shanghai 200000, China.
Sensors (Basel). 2014 Nov 18;14(11):21872-88. doi: 10.3390/s141121872.
A colloidal gold immunochromatographic assay (GICA) was developed for rapid detection of chloramphenicol (CAP) residues in aquatic products. A nitrocellulose (NC) membrane was used as the carrier, and the polyclonal CAP antibody was used as the marker protein. The average diameter of as-prepared colloidal gold nanoparticles (AuNPs) was about 20 nm. The optimal pH value of colloidal gold solutions and the amount of the antibody of CAP were 8.0 and 7.2 μg/mL, respectively. The CAP antibody was immobilized onto the conjugate pad after purification. The CAP conjugate and goat anti-rabbit IgG (secondary antibody) were coated onto the NC membrane. Next, the non-specific sites were blocked with 1% bovine serum albumin. The minimum detectable concentration of CAP in standard solution is 0.5 ng/mL, with good reproducibility. For the real samples from crucian carps injected with a single-dose of CAP in the dorsal muscles, the minimum detectable concentration of CAP residues was 0.5 µg/kg. The chromatographic analysis time was less than 10 min, and the strip had a long storage lifetime of more than 90 days at different temperatures. The strips provide a means for rapid detection of CAP residues in aquatic products.
建立了一种胶体金免疫层析法(GICA)用于快速检测水产品中氯霉素(CAP)残留。以硝酸纤维素(NC)膜为载体,以氯霉素多克隆抗体作为标记蛋白。所制备的胶体金纳米颗粒(AuNPs)的平均直径约为20nm。胶体金溶液的最佳pH值和氯霉素抗体用量分别为8.0和7.2μg/mL。氯霉素抗体经纯化后固定于结合垫上。将氯霉素偶联物和羊抗兔IgG(二抗)包被在NC膜上。接下来,用1%牛血清白蛋白封闭非特异性位点。标准溶液中氯霉素的最低检测浓度为0.5ng/mL,重现性良好。对于背部肌肉注射单剂量氯霉素的鲫鱼实际样品,氯霉素残留的最低检测浓度为0.5μg/kg。色谱分析时间小于10分钟,试纸条在不同温度下具有超过90天的长储存寿命。该试纸条为快速检测水产品中氯霉素残留提供了一种方法。