School of Medicine, Hebei University of Engineering, Handan, 056000, Hebei Province, China.
Handan Municipal Centre for Disease Control and Prevention, Handan, 056000, Hebei Province, China.
Appl Microbiol Biotechnol. 2020 Jul;104(14):6375-6383. doi: 10.1007/s00253-020-10699-5. Epub 2020 Jun 2.
The emergence of lactic acid bacteria (LABs) resistant to existing antimicrobial drugs is a growing health crisis. To decrease the overuse of antibiotics, molecular diagnostic systems that can rapidly determine the presence of antibiotic resistance (AR) genes in LABs from yogurt samples are needed. This paper describes a fully integrated, miniaturized plastic chip and closed-tube detection chemistry that performs multiplex nucleic acid amplification. High-throughput identification of AR genes was achieved through this approach, and six AR genes were analyzed simultaneously in < 2 h. This time-to-result included the time required for the extraction of DNA. The detection limit of the chip was 10 CFU mL, which was consistent with that of tube LAMP. We detected and identified multiple DNAs, including streptomycin, tetracycline, and vancomycin resistance-associated genes, with complete concordance to the Kirby-Bauer disk diffusion method.Key Points• A miniaturized chip was presented, and multiplex nucleic acid amplification was performed.• The device can be integrated with LAMP for rapid detection of antibiotic resistance genes.• The approach had a high throughput of AR gene analysis in lactic acid bacteria.
乳酸菌（LAB）对抗生素耐药性的出现是一个日益严重的健康危机。为了减少抗生素的过度使用，需要一种分子诊断系统，能够从酸奶样本中快速确定 LAB 中抗生素耐药（AR）基因的存在。本文介绍了一种完全集成的、微型化的塑料芯片和封闭式检测化学试剂，可进行多重核酸扩增。通过这种方法可以实现高通量的 AR 基因鉴定，并且可以在 <2 小时内同时分析 6 个 AR 基因。这个结果时间包括 DNA 提取所需的时间。该芯片的检测限为 10 CFU mL，与管环介导等温扩增（LAMP）一致。我们用该芯片检测和鉴定了多种 DNA，包括链霉素、四环素和万古霉素耐药相关基因，与 Kirby-Bauer 圆盘扩散法完全一致。
关键点
• 提出了一种微型化的芯片，并进行了多重核酸扩增。
• 该装置可以与 LAMP 集成，用于快速检测抗生素耐药基因。
• 该方法在乳酸菌中具有高通量的 AR 基因分析能力。
