中文摘要：

基于萤火虫萤光素酶生物发光反应体系，以SpectraMaxM5酶标仪为发光强度测试设备，建立了测定化学物对萤光素酶抑制毒性的微板发光测试新方法。系统地研究了萤光素酶浓度、萤光素浓度、ATP浓度、pH、温度、反应时间等实验条件对发光强度的影响。发现ATP对发光呈现双相响应关系，最大发光度出现在ATP浓度为1．1×10μmol·L-1。应用该方法成功地测定了NaF，NaCl，KBr和NaBF4对萤光素酶的发光抑制毒性效应，基于非线性最小二乘法拟合化学物对萤光素酶毒性的剂量一效应曲线（DRC），拟合效应与实验结果之间的决定系数（R2）均大于0．99。通过拟合的DRC参数，准确地计算了化学物的半数效应浓度EC50。对比有关文献方法，萤光素酶毒性测试的微板发光法具有更简便快捷，节省试剂药品，便于多次平行测定从而提高准确度等优点。

英文摘要：

A new microplate luminometry for the toxicity bioassay of chemicals on firefly luciferase, was developed using the multifunctional microplate reader （SpectraMax MS） to measure the luminous intensity of luciferase. Elects of luciferase concen- tration, luciferin concentration, ATP concentration, pH, temperature, and reaction time on the luminescence were systematical- ly investigated. It was found that ATP exerted a biphasic response on the luciferase luminescence and the maximum relative light units （RLU） occurred at an ATP concentration of 1.1 × 10^-4 mol . L-1. The method was successfully employed in the toxic effect test of NaF, NaC1, KBr and NaBF4 on luciferase. Using nonlinear least square technique, the dose-response curves （DRC） of the 4 chemicals were accurately fitted with the coefficient of determination （R2） between the fitted and observed responses be- ing greater than 0. 99. The median effective concentration （EC50） of the 4 chemicals were accurately measured from the DRC models. Compared with some literatures, the bioassay is a fast easy-operate and cost-effective method with high accuracy.