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采用胶体金免疫层析法快速检测水产品中氯霉素残留量

Rapid detection of chloramphenicol residues in aquatic products using colloidal gold immunochromatographic assay.

作者信息

Zhou Chennan, Zhang Xueyin, Huang Xinxin, Guo Xishan, Cai Qiang, Zhu Songming

机构信息

College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou 310058, China.

Shanghai Entry-Exit Inspection and Quarantine Bureau, Shanghai 200000, China.

出版信息

Sensors (Basel). 2014 Nov 18;14(11):21872-88. doi: 10.3390/s141121872.

Abstract

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.

摘要

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。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/64cc/4279566/76458bddea0a/sensors-14-21872f1.jpg

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