Liu Nan, Su Pu, Gao Zhixian, Zhu Maoxiang, Yang Zhihua, Pan Xiujie, Fang Yanjun, Chao Fuhuan
Institute of Hygiene and Environmental Medicine, Tianjin 300050, China.
Anal Chim Acta. 2009 Jan 19;632(1):128-34. doi: 10.1016/j.aca.2008.10.061. Epub 2008 Nov 5.
Suspension array technology for simultaneous detection of three kinds of veterinary drugs, chloramphenicol (CAP), clenbuterol and 17-beta-estradiol has been developed. Conjugates of chloramphenicol and clenbuterol coupled with bovine serum albumin were synthesized and purified. Probes of suspension array were constituted by coupling the three conjugates on the fluorescent microspheres/beads and the microstructures of the beads' surface were observed by scanning electron microscopy which was a direct confirmation for the successful conjugates' coupling. The optimal addition of conjugates and the amounts of antibodies were optimized and selected, respectively. Standard curves were plotted and the coefficient of determination-R(2) was greater than 0.989 which suggested good logistic correlation. The detection ranges for the three veterinary drugs are 40-6.25x10(5) ng L(-1), 50-7.81x10(5) ng L(-1) and 1x10(3-)7.29x10(5) ng L(-1), respectively and the lowest detection limits (LDLs) of them are 40, 50 and 1000 ng L(-1), respectively. The suspension array is specific and has no significant cross-reactivity with other chemicals. Meanwhile, unknown samples were detected by suspension array and ELISA in comparison with each other. The errors between found and real for the detection of the unknown samples were relatively small to both of the two methods, whereas, the detection ranges of suspension array are broader and sensitive than that of the traditional ELISA. The high-throughput suspension array is proved to be a novel method for multi-analysis of veterinary drugs with simple operation, high sensitivity and low cost.
已开发出用于同时检测三种兽药(氯霉素(CAP)、克伦特罗和17-β-雌二醇)的悬浮阵列技术。合成并纯化了与牛血清白蛋白偶联的氯霉素和克伦特罗偶联物。通过将三种偶联物偶联到荧光微球/微珠上构成悬浮阵列探针,并通过扫描电子显微镜观察微珠表面的微观结构,这是偶联物成功偶联的直接证据。分别优化并选择了偶联物的最佳添加量和抗体量。绘制了标准曲线,测定系数R²大于0.989,表明具有良好的逻辑相关性。三种兽药的检测范围分别为40 - 6.25×10⁵ ng L⁻¹、50 - 7.81×10⁵ ng L⁻¹和1×10³ - 7.29×10⁵ ng L⁻¹,它们的最低检测限(LDL)分别为40、50和1000 ng L⁻¹。该悬浮阵列具有特异性,与其他化学物质无明显交叉反应。同时,用悬浮阵列和酶联免疫吸附测定法(ELISA)相互比较检测未知样品。两种方法检测未知样品时,测定值与真实值之间的误差都相对较小,然而,悬浮阵列的检测范围比传统ELISA更宽且更灵敏。高通量悬浮阵列被证明是一种用于兽药多分析的新方法,操作简单、灵敏度高且成本低。