Monoclonal antibody produced by heterologous indirect enzyme-linked immunosorbent assay and its application for parathion residue determination in agricultural and environmental samples

A heterologous indirect enzyme-linked immunosorbent assay (ELISA) was developed, which was based on monoclonal antibody (Mab) to determine parathion residue in agricultural and environmental samples. Eight Mabs were produced by introducing the heterologous indirect ELISA into the screening procedure. It was shown that these Mabs had more broad-reactivity with twenty competitors than that of 5H7 (Mab produced by homologous screening). So it became much easier using these new Mabs to develop heterologous immunoassays. In addition, all the developed heterologous ELISAs could be used to determine parathion residue in semiquantitative or quantitative level, and their detection limitation (LOD) was around 2 ng/mL. Compared with the previous heterologous ELISA (hapten 11/5H7) with IC₅₀ of 13.3 ng/mL, a better heterologous ELISA (hapten 11/1E1) was obtained with IC₅₀ of 3.8 ng/mL, which improved the sensitivity 3.5 times. Finally, the latter was applied to parathion residue determination in spiked agricultural and environmental samples without extraction or cleanup. The average recoveries of parathion added to water, soil, cucumber, Chinese cabbage and carrot were between 80.4 % and 111.8 %. The LOD for water and soil samples was 5 ng/mL, and the LOD for cucumber, rice and corn samples was 10 ng/mL.     

Persistence of Bacillus thuringiensis deposits in a hardwood forest,after aerial application of a commercial formulation at two dosage rates

Abstract

A commercial formulation of Bacillus thuringiensis Berliner var. kurstaki (BTK), Foray® 48B, was sprayed aerially over four blocks B13, B14, B15A and B15B in an oak forest in Wayne County, Pennsylvania during May 1990. B13 and B14 were sprayed at 75 billion international units (BIU) in 5.91 litres/ha and the other two at 50 BIU in 3.94 litres/ha. Oak foliage was collected at different intervals of time after treatment. Three types of bioassays were conducted against fourth instar gypsy moth larvae, viz., direct feeding of sprayed foliage, feeding on diet containing homogenized foliage, and force‐feeding of foliar extracts. Larval mortalities were converted into international units of BTK activity per unit area (IU/cm²) of foliage. Foliar extracts were also subjected to enzyme‐linked immunosorbent assay (ELISA) to determine the concentration of delta‐endotoxin protein. Regardless of the type of bioassay used, bioactivity of BTK persisted in foliage for about a week in all the blocks. The half‐life of inactivation, DT₅₀, ranged from ca 12 to 22 h. The immunoassay data indicated a shorter duration of persistence (i.e., about 2 d) of the delta‐endotoxin protein, with DT₅₀ values ranging from 10 to 15 h. Formulation ingredients present in Foray 48B played a role in the toxicity of BTK to gypsy moth larvae.     

Production of the monoclonal antibody against tylosin for immunoassay of macrolide antibiotics in milk

The objective of the present study was to develop an immunoassay for multi-determination of the residues of four macrolide antibiotics in milk. Tylosin was derivatized with 6-aminohexanoic acid to synthesize a hapten that was used to produce the monoclonal antibody. The obtained monoclonal antibody simultaneously recognized tylosin, acetylisovaleryltylosin, tilmicosin and desmycosin with crossreactivities of 100%, 91%, 49% and 76%, respectively. Then an indirect competitive immunoassay was developed to determine the four analytes in milk simultaneously. The limits of detection for the four analytes were in the range of 5.5–11.7 ng/mL depending on the compound. The recoveries of the four analytes from fortified blank milk were in the range of 78%–96% with coefficients of variation lower than 11.4%. The developed immunoassay could be used as a practical tool for rapid screening the residues of the four anlytes in milk.     

基于荧光免疫法的地表水中2，4-D原位快速检测

采用荧光免疫法原位测定地表水中2,4-二氯苯氧乙酸(2,4-D),方法在7.2μg/L～211.7μg/L范围内线性良好,最低检出限为2.6 μg/L.对空白水、长江水和湖泊水样品做2,4-D的4个质量浓度水平的加标回收试验,7次测定结果的RSD为3.9％～5.8％,加标回收率为85.9％～106％.用该方法与国标法同时测定黄石市内某长江水源地和某湖泊水中的2,4-D,结果无明显差异.     

环境样品中多氯联苯的生物检测方法研究进展

多氯联苯(PCBs)是一种环境中广泛存在的典型的持久性有毒有机污染物.对人类和环境产生了许多有害影响.随着对多氯联苯的研究越来越广泛,其监测方法研究发展得也很快.本文综述了近些年来国内外采用的多种多氯联苯的分析方法.特别介绍了酶联免疫法、生物传感器法、荧光免疫PCR 等近年来发展迅速的检测技术的研究情况.比较了各种免疫方法的检测范围和检测灵敏度,指出了目前利用免疫方法检测多氯联苯存在的问题,并对免疫分析技术在多氯联苯检测中的发展前景进行了展望.     

关于多氯联苯测定的探讨

对PCB测定中所使用的Aroclor和单体标准品的区别、联系和使用进行了分析阐述,并对PCB测定中的气相色谱法、气相色谱质谱联用法、生物分析法、免疫分析法的优点、局限性进行了比较.对今后的PCB的测定提出了建议。     

乙腈和正己烷对环境特征污染物免疫传感分析的影响

利用前期研发的倏逝波全光纤免疫传感系统,使用间接竞争免疫模式对双酚A(bisphenol A,BPA)进行了定量分析,其检测灵敏度可达0.2μg·L~(-1),线性检测区间为0.3~33.4μg·L~(-1).重点考察了乙腈和正己烷两种常用有机溶剂对双酚A免疫传感分析的影响,初步探讨了有机溶剂对免疫传感分析的影响机制.结果表明,正己烷含量高达10%都不会对双酚A免疫传感分析产生明显影响,而乙腈的影响比较大.当乙腈含量较低时,可用于双酚A的定量检测;而当乙腈含量较高时,可完全抑制均相溶液中抗体抗原之间的亲和反应,不能用于双酚A的定量分析.其原因可能是乙腈取代了一部分结合在抗体表面的水分子,导致抗体构象改变或者抗体与抗原的亲和力变化而造成的.     

农药残留速测技术研究进展

本文综述了国内外农药快速检测技术的研究状况。在速测技术中 ,色 -质联用分析法已发展得比较成熟 ,所得结果比较可靠 ,但该法的测定时间仍较长 ;免疫分析法具有简单、快速、价廉、特异性高、需样品量少等优点 ;生物传感器法更方便、快捷。     

Application of a permethrin immunosorbent assay method to residential soil and dust samples

A low-cost, high throughput bioanalytical screening method was developed for monitoring cis/trans-permethrin in dust and soil samples. The method consisted of a simple sample preparation procedure [sonication with dichloromethane followed by a solvent exchange into methanol:water (1:1)] with bioanalytical detection using a magnetic particle enzyme-linked immunosorbent assay (ELISA). Quantitative recoveries (83–126 %) of cis/trans-permethrin were obtained for spiked soil and dust samples. The percent difference of duplicate ELISA analyses was within ± 20 % for standards and ± 35 % for samples. Similar sample preparation procedures were used for the conventional gas chromatography/mass spectrometry (GC/MS) analysis except that additional cleanup steps were required. Recoveries of cis/trans-permethrin ranged from 81 to 108 % for spiked soil and dust samples by GC/MS. The ELISA-derived permethrin concentrations were highly correlated with the GC/MS-derived sum of cis/trans-permethrin concentrations with a correlation coefficient (r) of 0.986. The ELISA method provided a rapid qualitative screen for cis/trans-permethrin in soil and dust while providing a higher sample throughput with a lower cost as compared to the GC/MS method. The ELISA can be applied as a complementary, low-cost screening tool to prioritize and rank samples prior to instrumental analysis for exposure studies.     

pH和铜离子对微囊藻毒素-LR荧光免疫检测的影响与消除

研究了pH和铜离子对倏逝波全光纤免疫传感器检测微囊藻毒素-LR（MC-LR）的影响及其消除措施.研究表明,过酸或过碱条件对MC-LR免疫检测均有较强烈的影响,当pH<6或pH>8时,系统检测的信号随pH的降低或增大明显下降;而当pH在6～8之间时,检测标准曲线的IC₅₀为1.01～1.04 μg/L,检测区间在0.12～10.5 μg/L之间,较适合MC-LR的检测.低浓度铜离子对MC-LR免疫检测的影响不大,当CuSO₄浓度>5 mg/Ｌ时,系统检测荧光信号明显下降,而当CuSO₄浓度达到10 mg/Ｌ时,系统检测信号下降70％以上.在预反应混合物中添加1%的螯合剂EDTA,能有效抑制铜离子对免疫检测的影响.