Zhang Liding, Wei Qiujiang, Han Qinqin, Chen Qiang, Tai Wenlin, Zhang Jinyang, Song Yuzhu, Xia Xueshan
Molecular Medicine Research Center of Yunnan Province, Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming, China.
Kunming Biomed International, Kunming, China.
Front Microbiol. 2018 Feb 6;9:94. doi: 10.3389/fmicb.2018.00094. eCollection 2018.
is an important human food-borne zoonosis bacterial pathogen, and can cause clinically severe diarrhea. There is an urgent need to develop a specific, sensitive, and rapid methodology for detection of this pathogen. In this study, loop-mediated isothermal amplification (LAMP) combined with magnetic immunocapture assay (IC-LAMP) was first developed for the detection of in pure culture, artificial milk, and clinical stool samples. This method exhibited a detection limit of 8.7 CFU/mL. Compared with polymerase chain reaction, IC-LAMP is sensitive, specific, and reliable for monitoring Additionally, IC-LAMP is more convenient, efficient, and rapid than ordinary LAMP, as it is more efficiently enriches pathogen cells without extraction of genomic DNA. Under isothermal conditions, the amplification curves and the green fluorescence were detected within 30 min in the presence of genomic DNA template. The overall analysis time was approximately 1 h, including the enrichment and lysis of the bacterial cells, a significantly short detection time. Therefore, the IC-LAMP methodology described here is potentially useful for the efficient detection of in various samples.
是一种重要的食源性人畜共患病细菌病原体，可导致临床上严重的腹泻。迫切需要开发一种特异性、灵敏且快速的方法来检测这种病原体。在本研究中，首次开发了环介导等温扩增（LAMP）结合磁免疫捕获测定法（IC-LAMP）用于检测纯培养物、人工牛奶和临床粪便样本中的。该方法的检测限为8.7 CFU/mL。与聚合酶链反应相比，IC-LAMP对于监测而言灵敏、特异且可靠。此外，IC-LAMP比普通LAMP更方便、高效且快速，因为它无需提取基因组DNA就能更有效地富集病原体细胞。在等温条件下，在存在基因组DNA模板的情况下，30分钟内即可检测到扩增曲线和绿色荧光。总体分析时间约为1小时，包括细菌细胞的富集和裂解，检测时间显著缩短。因此，本文所述的IC-LAMP方法对于高效检测各种样本中的可能具有潜在用途。
