四环素类抗生素酶联适体分析试剂盒的研制

Preparation of enzyme-linked aptamer assay kit for determining tetracycline antibiotics

研制了适用于检测动物源性食品中四环素类抗生素残留的竞争取代酶联适体分析试剂盒。以土霉素为靶标建立检测的标准曲线,结果表明:研制的试剂盒检测限为2. 3μg/L,线性检测范围在5～100μg/L。将四环素、土霉素、金霉素和强力霉素添加到猪肉、蜂蜜、牛奶样品中进行添加回收实验,并对试剂盒的精密度、特异性、稳定性和有效期进行测定。结果显示:四环素、土霉素、金霉素和强力霉素的添加回收率分别为77. 6%～88. 2%,82. 9%～93. 5%,78. 3%～86. 5%和86. 4%～92. 7%,供试样品检测结果的批内和批间变异系数均低于15%,符合兽药残留分析要求;该试剂盒对四环素类抗生素具有良好的特异性,能抵抗反复冻融60次以上,在4℃至少可保存6个月。

An enzyme-linked aptamer kit for competitive detection of tetracyclines residues in animal origin foods was developed.The standard curve of the test was established with oxytetracycline as the target,and the results showed that the detection limit of the kit was 2.3 g/L and the linear detection range was 5-100 μg/L.The recovery experiment was performed by adding the tetracycline,oxytetracycline,chlortetracycline and doxycycline to the samples of pork,honey and milk.The sensitivity,precision,specificity,stability and validity of the kit were measured,and the results showed that recoveries of tetracycline,oxytetracycline,chlortetracycline and doxycycline were77.6%-88.2%,82.9%-93.5%,78.3%-86.5% and 86.4%-92.7%,respectively.The coefficient of variation of intra-assay and inter-assay was less than 15%,which meets the analysis requirements of veterinary drug residues.The kit has good specificity for tetracycline antibiotics and good stability even after repeated freezing and thawing more than 60 times.The stability can be maintained under storage at 4℃ for at least six months.