食品科学 ›› 2019, Vol. 40 ›› Issue (14): 333-338.doi: 10.7506/spkx1002-6630-20181029-330

• 安全检测 • 上一篇    下一篇

高通量抗生素残留初筛试剂盒的研制及应用

范 维,高晓月,李贺楠,郭文萍,陈淑敏,孙 勇,李莹莹   

  1. 中国肉类食品综合研究中心,北京食品科学研究院,北京 100068
  • 出版日期:2019-07-25 发布日期:2019-07-23
  • 基金资助:
    “十三五”国家重点研发计划重点专项(2018YFC1603400);北京市优秀人才培养资助青年骨干个人项目(2017754154700G101)

Development and Application of Kit for High-Throughput Detection of Antibiotic Residues

FAN Wei, GAO Xiaoyue, LI Henan, GUO Wenping, CHEN Shumin, SUN Yong, LI Yingying   

  1. China Meat Research Center, Beijing Academy of Food Science, Beijing 100068, China
  • Online:2019-07-25 Published:2019-07-23

摘要: 以动物源性食品中常见的单一或配伍使用的抗生素为检测目标,开发基于微生物显色原理的高通量快速检测其残留的初筛试剂盒。选取嗜热脂肪芽孢杆菌(Bacillus sterothermophilus CICC 10392)作为指示菌,优化其微胶囊包埋条件,并固定于96 微孔板中,结合前期研究成果制成可同时对禽畜肉中常见的4 类抗生素进行联合初筛的试剂盒,克服了因菌种活化耗时较长及液体菌种运输困难等问题。结果表明:以0.1 g/mL聚乙烯醇为载体、pH 6.0的硼酸-磷酸盐溶液为交联剂对初始吸光度(A600 nm)为0.8的指示菌进行包埋,其机械强度、细胞活性及检测效果最好;该试剂盒对四环素类检出限为40~60 μg/kg,氨基糖苷类检出限为60~120 μg/kg,大环内酯类检出限为60~100 μg/kg,β-内酰胺类检出限为20~40 μg/kg,四环素与氨基糖苷类配伍使用检出限为20~40 μg/kg,β-内酰胺类与氨基糖苷类配伍使用检出限为10 μg/kg,符合国内外对抗生素残留限量的要求;用色谱法对试剂盒检测的70 份样品进行验证,无假阴性存在,说明该试剂盒结果可靠,可用于动物源性食品中抗生素残留的初筛检测。

关键词: 高通量, 抗生素残留, 包埋固定, 动物源性食品

Abstract: In this study, a rapid high-throughput screening kit for the detection of mono- and multi-residues of common antibiotics in animal-derived foods was developed based on the microbial chromogenic assay. Bacillus sterothermophilus (CICC 10392) was used as the indicator strain. Its cells were immobilized under optimized conditions and seeded into the wells of a 96-well microplate to detect four types of common antibiotic residues in livestock and poultry meat. As a result, the problem of time-consuming cultivation and the difficulty of transporting liquid bacterial culture could be overcome. The results showed that use of 0.1 g/mL polyvinyl alcohol as the carrier, boric acid-phosphate solution (pH 6.0) as the crosslinking agent, and an initial bacterial suspension concentration (A600 nm) of 0.8 were the best immobilization conditions in terms of the mechanical stability and microbial activity. The detection limits for tetracyclines, aminoglycosides, macrolides, β-lactams separately, and aminoglycosides in combination with tetracyclines or β-lactams were 40–60, 60–120, 60–100, 20–40, 20–40, and 10 μg/kg, respectively, conforming to the requirements for antibiotic residue limits. The same results were obtained on 70 animal-derived food samples using the kit and chromatography. The results indicated that the kit could be used to reliably detect antibiotic residues in animal-derived food samples.

Key words: high-throughput, antibiotic residues, immobilization, animal-derived food

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