Department of Analytical Chemistry, Faculty of Chemistry, University of Tabriz, Tabriz, 51666 16471, Iran.
Pharmaceutical Analysis Research Center and Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran.
Mikrochim Acta. 2023 Mar 4;190(4):112. doi: 10.1007/s00604-023-05687-1.
For sensitive detection of the L-fuculokinase genome related to the Haemophilus influenzae (H. influenzae), this research work demonstrates the label-free electrochemical-based oligonucleotide genosensing assay relying on the performing hybridization process. To enhance the electrochemical responses, multiple electrochemical modifier-tagged agents were effectively utilized. For attaining this goal, NiCr-layered double hydroxide (NiCr LDH) has been synthesized and combined with biochar (BC) to create an efficient electrochemical signal amplifier that has been immobilized on the surface of the bare Au electrode. Low detection and quantification limits (LOD and LOQ) associated with the designed genosensing bio-platform to detect L-fuculokinase have been achieved to 6.14 fM and 11 fM, respectively. Moreover, the wide linear range of 0.1 to 1000 pM demonstrates the capability of the designed platform. Investigated were the 1-, 2-, and 3-base mismatched sequences, and the negative control samples clarified the high selectivity and better performance of the engineered assay. The values of 96.6-104% and 2.3-3.4% have been obtained for the recoveries and RSDs, respectively. Furthermore, the repeatability and reproducibility of the associated bio-assay have been studied. Consequently, the novel method is appropriate for rapidly and quantitatively detecting H. influenzae, and is considered a better candidate for advanced tests on biological samples such as urine samples.
为了敏感地检测与流感嗜血杆菌(Haemophilus influenzae,H. influenzae)相关的 L-岩藻糖激酶基因组,本研究工作展示了一种基于无标记电化学的寡核苷酸基因传感分析方法,该方法依赖于执行杂交过程。为了增强电化学响应,有效地利用了多种电化学修饰剂标记的试剂。为了实现这一目标,合成了 NiCr 层状双氢氧化物(NiCr LDH)并与生物炭(BC)结合,创建了一种有效的电化学信号放大器,将其固定在裸 Au 电极表面。所设计的基因传感生物平台用于检测 L-岩藻糖激酶的检测和定量下限(LOD 和 LOQ)分别达到 6.14 fM 和 11 fM。此外,设计平台的线性范围为 0.1 至 1000 pM,显示出其能力。研究了 1、2 和 3 个碱基错配序列,而阴性对照样品则表明了工程化测定的高选择性和更好性能。回收率和 RSD 的值分别为 96.6-104%和 2.3-3.4%。此外,还研究了相关生物测定的重复性和重现性。因此,该新方法适用于快速定量检测 H. influenzae,并且被认为是对尿液等生物样本进行高级测试的更好候选者。