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化学激活荧光素酶基因表达检测法(CALUX生物测定法):一种用于筛选二噁英类化合物的经济高效的快速筛选技术。

CALUX bioassay: a cost-effective rapid screening technique for screening dioxins like compounds.

作者信息

Sakthivel Selvaraj, Balasubramanian Prithiviraj, Nakamura Masafumi, Ko Shunkei, Chakraborty Paromita

出版信息

Rev Environ Health. 2016 Mar;31(1):149-52. doi: 10.1515/reveh-2015-0078.

Abstract

Xenobiotic detection systems-chemically activated luciferase expression (XDS-CALUX) bioassay in determining the toxic equivalency (TEQ) of PCDD/Fs from contaminated sites reported in several papers has been discussed in this study. CALUX bioassay method has been validated by an effective combined column clean-up system followed by addition of samples to monolayer cell cultures of H1L6.1c3 cell line in 96 well plates. Cultures are then examined under microscope after 24 h incubation followed by rinsing with 75 μL phosphate buffer saline and 30 μL of cell culture lysis. The response is observed in the luminometer and expressed in relative light unit (RLUs). CALUX-TEQ is estimated from a TCDD standard curve for unknown samples. Quality control in CALUX is done by selecting the range of CALUX values falling in the center of the linear standard curve. For developing nations CALUX biossay can be used as a cost effective and rapid screening technique for screening xenobiotic compounds from the hotspots like open solid waste burning sites, informal e-waste recycling workshops and industrial zones where constant monitoring for such compounds is required.

摘要

本研究讨论了多篇论文中报道的用于测定污染场地多氯二苯并对二噁英/多氯二苯并呋喃(PCDD/Fs)毒性当量(TEQ)的异生物质检测系统——化学激活荧光素酶表达(XDS-CALUX)生物测定法。CALUX生物测定法已通过有效的组合柱净化系统进行验证,随后将样品添加到96孔板中H1L6.1c3细胞系的单层细胞培养物中。孵育24小时后,在显微镜下检查培养物,然后用75μL磷酸盐缓冲盐水冲洗,再加入30μL细胞培养裂解液。在发光计中观察反应,并以相对光单位(RLUs)表示。根据未知样品的2,3,7,8-四氯二苯并对二噁英(TCDD)标准曲线估算CALUX-TEQ。CALUX中的质量控制是通过选择落在线性标准曲线中心的CALUX值范围来完成的。对于发展中国家,CALUX生物测定法可作为一种经济高效的快速筛选技术,用于从露天固体废物焚烧场、非正规电子废物回收车间和工业区等热点地区筛选异生物质化合物,这些地区需要对这类化合物进行持续监测。

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