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全细胞纸条生物传感器用于半定量环境基质中的四环素类抗生素。

Whole-cell paper strip biosensors to semi-quantify tetracycline antibiotics in environmental matrices.

机构信息

Institute of Organic Contaminant Control and Soil Remediation, College of Resources and Environmental Sciences, Nanjing Agricultural University, Nanjing, 210095, PR China.

Department of Environment and Geography, University of York, Heslington, York, YO10 4DU, UK.

出版信息

Biosens Bioelectron. 2020 Nov 15;168:112528. doi: 10.1016/j.bios.2020.112528. Epub 2020 Aug 27.

Abstract

A novel, low-cost, and portable paper strip biosensor was developed for the detection of tetracycline antibiotics. Escherichia coli/pMTLacZ containing the tetracycline-mediated regulatory gene used as recognition elements with β-galactosidase as the reporter protein was designed and applied to cheap and portable Whatman filter paper as the carrier to prepare this paper strip biosensor. The detection process was optimized by using EDTA and polymyxin B as a sensitizer to improve the accuracy of detection for complicated matrices. The paper strip biosensor was suitable for tetracycline concentrations in the range of 75-10000 μg/L in water and 75-7500 μg/L in soil extracts. Detection limits of 5.23-17.1 μg/L for water and 5.21-35.3 μg/kg for the EDTA soil extracts were achieved at a response time of 90 min. The standard deviation (SD) of detected values by the biosensor paper strip compared to those determined by HPLC was between 13.4 and 59.6% for tetracycline and 2.01-33.5% for oxytetracycline in water and was between 6.22 and 72.8% for tetracycline and 5.90-43.4% for oxytetracycline in soil. This suggests that the paper strip biosensor was suitable for detecting both tetracycline and oxytetracycline in water, and could provide a suitable detection for extractable oxytetracycline in soils. Therefore, this biosensor provides a simple, economical, and portable piece of field kit for on-site monitoring of tetracyclines in a variety of environmental samples, such as pond water and agricultural soil that are susceptible to tetracycline pollution from feed additives and fertilization with livestock manure.

摘要

一种新型的、低成本的、便携式的纸-strip 生物传感器被开发出来,用于检测四环素类抗生素。以四环素介导的调控基因作为识别元件,以β-半乳糖苷酶作为报告蛋白,设计了含有该基因的大肠杆菌/pMTLacZ,并将其应用于廉价且便携的 Whatman 滤纸作为载体,制备了这种纸-strip 生物传感器。通过使用 EDTA 和多粘菌素 B 作为增敏剂来优化检测过程,以提高复杂基质检测的准确性。该纸-strip 生物传感器适用于水和土壤提取物中四环素浓度分别为 75-10000μg/L 和 75-7500μg/L 的范围。在 90 分钟的响应时间内,水的检测限为 5.23-17.1μg/L,EDTA 土壤提取物的检测限为 5.21-35.3μg/kg。与 HPLC 相比,该生物传感器检测值的标准偏差(SD)在水中为四环素的 13.4-59.6%,土霉素的 2.01-33.5%,在土壤中为四环素的 6.22-72.8%,土霉素的 5.90-43.4%。这表明,该纸-strip 生物传感器适用于检测水中的四环素和土霉素,并且可以为土壤中可提取的土霉素提供合适的检测。因此,该生物传感器为现场监测池塘水和农业土壤等易受饲料添加剂和牲畜粪便施肥四环素污染的各种环境样品中的四环素提供了一种简单、经济、便携的现场检测试剂盒。

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