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A novel fluorescence based assay for the detection of organophosphorus pesticide exposed cholinesterase activity using 1-naphthyl acetate
Biochimie ( IF 3.3 ) Pub Date : 2019-02-26 , DOI: 10.1016/j.biochi.2019.02.014
Sheemona Chowdhary , Rajasri Bhattacharyya , Dibyajyoti Banerjee

Acetylcholinesterase (AChE) is the primary target of organophosphorus pesticides (OPs). Ellman's method using Acetylthiocholine (ATCh) is the standard approach for the detection of AChE activity. Though ATCh is a popular substrate, it has certain drawbacks as well. Because of these limitations, there is a need for the development of reliable and rapid assays for determination of AChE activity in cases of OP poisoning. In the present work, we have used 1-Naphthyl acetate (1-NA) as a fluorogenic substrate for the estimation of AChE activity of human erythrocytes. It is well known that due to inter-individual variation in AChE activity, the baseline value cannot be correctly predicted. Therefore, using 1-NA, we have developed a rapid, sensitive and baseline free assay for the estimation of AChE activity. The assay is based on reactivation and fluorescence quenching using a cocktail of oximes for the determination of cholinesterase activity in a post-exposure sample. Moreover, it is free from interference due to oximolysis which is an established limitation of ATCh. We feel that such an assay using 1-NA has the potential to be explored at the point of care for rapid detection of OP poisoning.



中文翻译:

一种新颖的基于荧光的测定方法,使用乙酸1-萘酯检测有机磷农药暴露的胆碱酯酶活性

乙酰胆碱酯酶(AChE)是有机磷农药(OPs)的主要目标。使用乙酰硫胆碱(ATCh)的Ellman方法是检测AChE活性的标准方法。尽管ATCh是一种流行的底物,但它也具有某些缺点。由于这些限制,需要开发可靠和快速的测定方法,以测定OP中毒时的AChE活性。在目前的工作中,我们已使用乙酸1-萘酯(1-NA)作为荧光底物来评估人红细胞的AChE活性。众所周知,由于AChE活性之间存在个体差异,因此无法正确预测基线值。因此,使用1-NA,我们已经开发了一种快速,灵敏和无基线的测定方法,用于评估AChE活性。该测定基于使用肟混合物的再活化和荧光猝灭,用于测定暴露后样品中的胆碱酯酶活性。此外,它不受由于氧肟分解引起的干扰,而这是ATCh的既定局限性。我们认为使用1-NA的这种测定方法有可能在快速检测OP中毒的护理点上进行探索。

更新日期:2019-02-26
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