【摘要】 目的:用自制重金属铜单抗和合成铜完全抗原,建立检测重金属铜离子的间接竞争化学发光酶联免疫（CLEIA）法,为植物中草药和土壤环境中铜的快速定量测定提供简便、可靠的方法。方法:使用本实验室前期制备并鉴定合格的优质铜单抗和合成铜抗原,建立铜间接竞争CLEIA法,并对其准确度、精密度、特异性和稳定性进行评价;采集植物中草药和土壤样品共31份,通过微波消解联合沉淀法进行前处理并制备待测样,由间接竞争CLEIA法进行铜含量测定,以ICP-AES仪器法所测结果为对照评价其可靠性。结果:该方法检测灵敏度为17. 9 ng/ml,检测限为1. 9 ng/ml,线性范围为3. 0～200 ng/ml;准确度试验回收率为98. 8%～105. 5%,精密度试验RSD≤5. 0%;特异性试验仅与Mg²⁺、Zn²⁺有13. 3%、3. 3%的交叉反应;酶标板置-20℃避光保存6周内其检测结果准确性未出现明显改变。对植物中草药和土壤样品进行前处理并制样检测,对比ICP-AES法检测结果,相关系数分别为0. 972和0. 989,两种方法具有较好相关性,表明间接竞争CLEIA法可应用于一般环境样品中铜的简便快速定量测定。结论:成功建立铜间接竞争CLEIA检测法,其灵敏度及各参数均符合国家法定相关样品铜离子检测技术指标的要求,可应用于一般环境样品中铜含量的快速测定。更多还原

【Abstract】 Objective: Use the prepared anti copper McAb and the synthesized copper antigen to establish the indirect competitive chemiluminescence linked enzyme immunoassay（ CLEIA） for simple,fast and quantitative detection of copper content in herbal plants and soil samples. Methods: Use the anti copper McAb and antigen to establish the indirect competitive CLEIA,and evaluate its accuracy,precision,specificity and stability performance. A total of 31 samples include herbal plants and soil were collected and pretreated by microwave digestion combined with precipitation. The copper content was determined by indirect competition CLEIA and ICPAES after sample preparation,and the relevance of the two methods was evaluated. Results: The copper detection sensitivity was 17. 9 ng/ml,the low detection limit was 1. 9 ng/ml,and the linear range was 3. 0 to 200 ng/ml. The recovery rate of accuracy test was 98. 8%to 105. 5%,and the RSD was less or equal to 5%. Specific test showed that the anti copper McAb had a slight cross reactivity rate of13. 3%and 3. 3%to Mg²⁺and Zn²⁺. Storage the CLEIA microplates at-20℃ away from light for 2 to 8 weeks,the stability test showed no significant change in 6 weeks. Herbal plants and soil samples were pre-treated and prepared into test samples detected by indirect competitive CLEIA and ICP-AES for copper content. The two methods showed good relativity with the correlation coefficient value of0. 972 and 0. 989. The above results indicated that the indirect competitive CLEIA method can be applied to the detection of copper content in various environmental samples. Conclusion: Successfully established the indirect competitive CLEIA for simple,fast and quantitative detection of copper content in various environmental samples,and the detection sensitivity and other parameters are in line with the requirements of national inspection technical.更多还原