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目的建立一种在体外快速测定乙酰胆碱酯酶(AChE)活性的方法,为有机磷毒剂中毒的诊断及救治提供依据。方法用改良的5,5-连硫代-双-2-硝基苯甲酸(DTNB)法测定AChE活性,以兔全血作为酶源进行方法学研究,并用不同种属动物全血进行了适用性考察,用SPECTRA MAX 190读板机测定412 nm处光密度的变化即可测定AChE活性。结果不同动物全血40倍稀释后作为酶源较为合适,批内差异为0.44%～0.67%,批间差异为1.03%～2.27%,该方法适用于兔、比格犬、豚鼠和猴全血AChE测定。梭曼和沙林对兔全血AChE活性抑制具有明显的量效关系。结论该方法简便、易行,适用于不同种属动物全血AChE活性的测定。
Objective To establish a rapid in vitro determination of acetylcholinesterase (AChE) activity of the method for the diagnosis and treatment of organophosphate poisoning provide the basis. Methods The activity of AChE was determined by a modified 5,5-dithiobis-2-nitrobenzoic acid (DTNB) method and the whole rabbit blood was used as the enzyme source for the methodological study. For sexual investigation, AChE activity was measured by measuring the change in optical density at 412 nm using a SPECTRA MAX 190 plate reader. Results The results showed that 40% dilution of whole blood of different animals could be used as enzyme source. The difference between batches was 0.44% -0.67%, and the difference between batches was 1.03% -2.27%. The method was suitable for rabbit, beagle dog, guinea pig and monkey AChE assay. Soman and sarin had a significant dose-response relationship with inhibition of AChE activity in whole blood of rabbits. Conclusion The method is simple and easy to perform and is suitable for the determination of AChE activity in whole blood of different species of animals.