Byzova Nadezhda A, Smirnova Nataliya I, Zherdev Anatoly V, Eremin Sergei A, Shanin Ilya A, Lei Hong-Tao, Sun Yuanming, Dzantiev Boris B
A.N. Bach Institute of Biochemistry, Russian Academy of Sciences, Leninsky prospect 33, 119071 Moscow, Russia.
A.N. Bach Institute of Biochemistry, Russian Academy of Sciences, Leninsky prospect 33, 119071 Moscow, Russia; Faculty of Chemistry, M.V. Lomonosov Moscow State University, Leninskie gory 1, building 3, 119991 Moscow, Russia.
Talanta. 2014 Feb;119:125-32. doi: 10.1016/j.talanta.2013.10.054. Epub 2013 Oct 30.
An immunochromatographic assay was developed to detect fluoroquinolone antibiotic ofloxacin based on the competitive binding of ofloxacin and the membrane-immobilized ofloxacin-protein conjugate to colloidal gold-labeled antibodies in the course of the labeled antibodies, and to test sample flow through the membrane. The specific feature of labeling by colloidal gold is that native antiserum is used instead of purified immunoglobulins or specific antibodies. This makes the synthetic procedure easier, with no sacrifice in the detection limit. The proposed test makes it possible to detect down to 30 ng mL(-1) of ofloxacin, which corresponds to the demands of food safety assessment. The assay time is 10 min. The assay provides reliable information on the ofloxacin content in milk without the sample preparation and in chicken and pork meat with the minimum sample preparation (the separation of the insoluble fraction of the homogenate by centrifugation). The high degree of detection of ofloxacin in foodstuffs by the proposed assay (70-112%) was shown by a comparison with the data obtained with the use of a commercial immunoenzymatic kit.
开发了一种免疫层析测定法，用于检测氟喹诺酮类抗生素氧氟沙星。该方法基于在标记抗体过程中，氧氟沙星与膜固定的氧氟沙星 - 蛋白质偶联物竞争结合胶体金标记抗体，并使测试样品流经该膜。胶体金标记的独特之处在于使用天然抗血清而非纯化的免疫球蛋白或特异性抗体。这使得合成过程更简便，且检测限不受影响。所提出的测试方法能够检测低至30 ng mL(-1)的氧氟沙星，符合食品安全评估的要求。检测时间为10分钟。该测定法无需样品制备即可提供牛奶中氧氟沙星含量的可靠信息，对于鸡肉和猪肉，只需进行最少的样品制备（通过离心分离匀浆中的不溶部分）。与使用商业免疫酶试剂盒获得的数据相比，所提出的测定法对食品中氧氟沙星的检测率较高（70 - 112%）。
