Abstract
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 103 CFU mL−1, 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. |
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This study was supported by the provincial Key Research and Development Program of Hebei (17275505D).
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ZJ, GD, and WL wrote the manuscript. ZJ, GD, and GY designed the chip and conducted experiments. WL conceived and designed research. GD and GY analyzed data. GL, WZ, and LY reviewed and edited the manuscript. All authors read and approved the manuscript.
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Jin, Z., Ding, G., Yang, G. et al. Rapid detection of antibiotic resistance genes in lactic acid bacteria using PMMA-based microreactor arrays. Appl Microbiol Biotechnol 104, 6375–6383 (2020). https://doi.org/10.1007/s00253-020-10699-5
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DOI: https://doi.org/10.1007/s00253-020-10699-5