摘要
以6-（1-萘氧基甲酰胺基）己酸（CNH）偶联牛血清白蛋白（BSA）作为免疫原（CNH-BSA）制备高特异性抗体,分别以CNH、4-（1-萘氧基甲酰胺基）丁酸（CNB）、3-（1-萘氧基甲酰胺基）丙酸（CNA）偶联卵清蛋白（OVA）得到包被抗原（CNH-OVA,CNB-OVA,CNA-OVA）,以上述抗体及包被原作为核心材料,研究了同源包被与异源包被模式及ELISA各影响因素对检测灵敏度的影响,建立了甲萘威异源间接竞争酶联免疫法,并考察了此方法对测定大米及土壤中甲萘威残留的适用性。结果表明：以CNA-OVA异源包被的间接竞争ELISA法具有较高的灵敏度,IC50为（10.51±0.11）μg·L-1,该方法的检测范围为2.07～47.30μg·L-1（以IC20～IC80为标准）。以0.5,1.0,2.0,4.0 mg·kg-1作为加标浓度,甲萘威在大米中的加标回收率为92.3%～111.6%,土壤中的加标回收率为85.3%～103.2%,相对标准偏差均在10%以内。与HPLC法的比对验证结果表明,ELISA和HPLC两种方法的分析结果无显著性差异（P＞0.05）。
The highly specific antibody was prepared with CNH（ 6-[（ 1-naphthyloxy） carbonyl]amino hexanoic acid） coupled with BSA（ bovine serum albumin） as immunogen. Three coating antigens,CNH-OVA,CNB-OVA and CNA-OVA were obtained with CNH（ 6-[（ 1-naphthyloxy）carbonyl] amino hexanoic acid）,CNB（ 4-[（ 1-naphthyloxy） carbonyl] amino butyric acid） and CNA（ 3-[（ 1-naphthyloxy） carbonyl]amino propanoic acid） coupled with OVA,respectively. Taking antibody and coating antigens mentioned above as key materials,the influences of homologous coating antigen,heterologous coating antigen and each ELISA influence factor on detection sensitivity were investigated,the indirect competitive ELISA of carbaryl was established,and the method＇s applicability to the measurement of carbaryl residues in rice and its soil was studied. The results showed that the sensitivity of the ELISA assay with CNA-OVA heterologous coating was high,and the IC50was（ 10. 51 ± 0. 11） μg·L-1. The detection range of ELISA was in the range of 2. 07-47. 30 μg·L-1（ IC20-IC80as a standard）. Taking 0. 5,1. 0,2. 0,4. 0 mg·kg-1as additive concentration,the spiked recoveries of carbaryl in the rice and soil samples were in the ranges of 92. 3%-111. 6%and 85. 3%-103. 2%,respectively. The relative standard deviations were less than 10%. There were no distinct difference between the results of ELISA and HPLC（ P 〉 0. 05）.
出处
《分析测试学报》
CAS
CSCD
北大核心
2014年第6期628-634,共7页
Journal of Instrumental Analysis
基金
公益性行业(农业)科研专项经费项目(201203094-2)
