用蚊虫酯酶单克隆抗体进行库蚊有机磷抗性的免疫学检测

IMMUNOLOGICAL DETECTION OF ORGANOPHOSPHATE RESISTANCE OF CULEX MOSQUITOES USING ANTI ESTERASE MONOCLONAL ANTIBODY

对不同品系致倦库蚊或淡色库蚊分别用抗致倦库蚊酯酶单克隆抗体进行ｄｏｔ－ＥＬＩＳＡ和夹心法ＥＬＩＳＡ测定，并与生物测试法和生化微量板法相比较。免疫学检测发现的抗性程度高于后两法，发现的抗性频率高于生化法。ＥＬＩＳＡ方法诊断抗性的阈值为ＯＤ４５０≥０．５，生化法阈值为β－Ｎ浓度≥２．５×１０－３μｍｏｌ／ｍｉｎ·ｍｇ蛋白。ｄｏｔ－ＥＬＩＳＡ法适用于现场快速筛选抗性，对高酯酶活力抗性品系的检测优于夹心法ＥＬＩＳＡ。

Different strains of Culex mosquitoes(Cx.pipiens quinquefasciatus and Cx.Pipiens pallens)were immunologically detected for organophosphate resistance with anti esterase monoclonal antibody,and the results were compared with those detected by bioassay and biochemical microplate assay.It was found that the resistance detection rate detected by immunologic methods were higher than the corresponding data detected by the biochemical method,and the levels of resistance detected by sandwich ELISA were higher than the corresponding levels detected by bioassay and the biochemical method.The thresholds for resistance in sandwich ELISA were (at absorbence 450)≥0.5,and in microplate assay were (at absorbence 550)≥2.5×10 3 μmol/min·mg protein. Dot ELISA method was developed to meet the requirement of the field test and proved to be fast and convenient,especially in the detection of samples with higher esterase activity.