Ultrasensitive detection of pathogenic bacteria by primer exchange reaction coupled with PGM

文献类型： 外文期刊

第一作者： Yu, Songling

作者： Yu, Songling;Xue, Lulu;Li, Xue;Wang, Xiuran;Bu, Shengjun;Zhou, Hongyu;Wan, Jiayu

作者机构：

关键词： Detection; Invertase; Personal glucose meter; Primer exchange reaction

期刊名称：MICROCHEMICAL JOURNAL （ 影响因子：4.8； 五年影响因子：4.5 ）

ISSN： 0026-265X

年卷期： 2023 年 187 卷

页码：

收录情况： SCI

摘要： This study proposes an innovative approach for highly sensitive and selective detection of pathogenic bacteria based on primer exchange reaction (PER) coupled with a personal glucose meter (PGM). The modified nucleic acid capture probe (CP, 3'-end modified biotin) was immobilized onto the surface of magnetic beads (MBs) and hybridized with the specific aptamer of Escherichia coli O157:H7 (AP) to form a double-stranded structure on the surface of MBs. The AP preferentially bound to E. coli O157:H7, thereby exposing some CP. Further, CP hy-bridized with a 5'-end sequence of long single-stranded DNA, which was autonomously synthesized by PER, and captured a nucleic acid reporter probe (5'-end modified biotin) via nucleic acid hybridization. Subsequently, numerous SA-labeled invertase conjugates were added. Next, glucose was produced on adding the substrate (sucrose) of the invertase, which was detected using PGM. Under optimal conditions, the experimental results showed the corresponding detection of E. coli O157:H7 with limits as low as 10 CFU/mL and linear ranges 10-107 CFU/mL. The proposed biosensor was used to detect pathogenic bacteria in milk samples successfully.

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