CAJ | 학술논문

为快速有效监控蔬菜（油菜薹）醚菊酯农药残留，采用醚菊酯与牛血清白蛋白偶联物（ ethofenprox -BSA）抗原免疫新西兰大白兔获得抗醚菊酯多克隆抗体。以醚菊酯与卵清白蛋白偶联物（ethofenprox - OVA）为检测抗原，利用此抗体，进行间接竞争 ELISA 法试验。通过棋盘格实验确定了最佳抗原包被反应质量浓度为0.25μg/mL，最佳抗原包被条件为37℃孵育2h；一抗最佳稀释度为1：32000；最佳 pH 值与离子浓度分别为6.0和0.16mol/L；最佳封闭试剂与有机溶剂分别为脱脂奶粉和10%甲醇/ PBS。在此优化条件下，建立了醚菊酯竞争抑制曲线，抗体的灵敏度（IC50）为0.28μg/ mL，检测线性范围（IC20～ IC80）为0.0798～3.578μg/ mL。采用该方法对市场抽样进行加标回收测定，平均回收率在75.26%～97.56%之间，实际样品检测间接 ELISA 与 GC/ MS 检测结果相关性系数达到0.994，灵敏度、精密度和准确度均符合快速检测要求。该法为快速、简便、低耗的醚菊酯免疫检测方法提供了可能。
위쾌속유효감공소채（유채대）미국지농약잔류，채용미국지여우혈청백단백우련물（ ethofenprox -BSA）항원면역신서란대백토획득항미국지다극륭항체。이미국지여란청백단백우련물（ethofenprox - OVA）위검측항원，이용차항체，진행간접경쟁 ELISA 법시험。통과기반격실험학정료최가항원포피반응질량농도위0.25μg/mL，최가항원포피조건위37℃부육2h；일항최가희석도위1：32000；최가 pH 치여리자농도분별위6.0화0.16mol/L；최가봉폐시제여유궤용제분별위탈지내분화10%갑순/ PBS。재차우화조건하，건립료미국지경쟁억제곡선，항체적령민도（IC50）위0.28μg/ mL，검측선성범위（IC20～ IC80）위0.0798～3.578μg/ mL。채용해방법대시장추양진행가표회수측정，평균회수솔재75.26%～97.56%지간，실제양품검측간접 ELISA 여 GC/ MS 검측결과상관성계수체도0.994，령민도、정밀도화준학도균부합쾌속검측요구。해법위쾌속、간편、저모적미국지면역검측방법제공료가능。
For rapid and accurate ELISA detection of ethofenprox residue in rapeseed stalks( vegetable), polyclonal antibody was obtained from rabbits with ethofenprox - bovine serum albumin(BSA)conjugate(ethofen-prox - BSA). Based on the antibody,an indirect competitive ELISA was developed with ethofenprox - OVA as the envelope antigen. Assays were performed in the ethofenprox - BSA(0. 25μg / mL)coated ELISA format in which the antibody was 1: 32 000 diluted. The optimized physicochemical factors(pH,ionic strength and blocking solu-tion)in the performance were obtained:blocking reagent was skim milk powder,pH6. 0,and ionic strength was 0. 16mol/ L. Calibration curve was established under the optimal conditions,from which the sensitivity(IC50 )was 0. 28μg / mL of antibody and the linear working range were IC20 - IC80 of 0. 079 8 - 3. 578μg / mL. Ethofenprox pes-ticides residue in rapeseed stalks collected from markets were screened by the developed ELISA with average recov-ery of 75. 26% - 97. 56% . The correlation coefficient of results obtained by both indirect competitive ELISA and GC / MS method was 0. 994. It concluded a practical ELISA method,and the validation of ethofenprox detection by ELISA and GC - MS in rapeseed stalks was proven.