Institute of Analytical Chemistry, Chemo- and Biosensors, University of Regensburg, 93053 Regensburg, Germany.
Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem 9190401, Israel.
Sensors (Basel). 2024 Oct 11;24(20):6549. doi: 10.3390/s24206549.
The continuous emergence of new illegal compounds, particularly psychoactive chemicals, poses significant challenges for current drug detection methods. Developing new protocols and kits for each new drug requires substantial time, effort, and dedicated manpower. Whole-cell bacterial bioreporters have been proven capable of detecting diverse hazardous compounds in both laboratory and field settings, identifying not only single compounds but also chemical families. We present the development of a microbial bioreporter for the detection of cocaine, the nervous system stimulant that is the second-most widely used illegal drug in the US. was transformed with a plasmid containing a bacterial bioluminescence gene cassette, activated by a cocaine-responsive signaling cascade. The engineered bioreporter is demonstrated to be a sensitive and specific first-generation detection system for cocaine, with detection thresholds of 17 ± 8 μg/L and 130 ± 50 μg/L in a buffer solution and in urine, respectively. Further improvement of the sensor's performance was achieved by altering the nucleotide sequence of the gene promoter, the construct's sensing element, using accelerated site-directed evolution. The applicability of ready-to-use paper strips with immobilized bioreporter cells was demonstrated for cocaine detection in aqueous solutions.
新的非法化合物不断涌现，特别是精神活性化学品，这给当前的毒品检测方法带来了重大挑战。为每种新毒品开发新的方案和试剂盒需要大量的时间、精力和专门的人力。全细胞细菌生物传感器已被证明能够在实验室和现场环境中检测到各种危险化合物，不仅能检测单一化合物，还能检测化学家族。我们开发了一种用于可卡因检测的微生物生物传感器，可卡因是美国第二大最广泛使用的非法药物，属于神经系统兴奋剂。将含有细菌生物发光基因盒的质粒转化为，该基因盒受可卡因反应信号级联激活。结果表明，该工程生物传感器是一种灵敏和特异的可卡因第一代检测系统，在缓冲溶液和尿液中的检测阈值分别为 17 ± 8 μg/L 和 130 ± 50 μg/L。通过使用加速的定向进化改变 基因启动子的核苷酸序列，即构建物的感应元件，进一步提高了传感器的性能。使用固定化生物传感器细胞的即用型纸条证明了其在水溶液中可卡因检测的适用性。
