A novel method for visual microarray detection of antibiotic resistance genes

Abstract

Antibiotic resistance genes (ARGs) are emerging pollutants that pose significant threats to both the environment and human health. Rapid detection of ARGs is essential for monitoring their levels and controlling their spread. However, traditional detection methods are often time-consuming and require specialized equipment, leading to delays. To address this issue, this study developed a novel method for visual microarray detection of ARGs, including sul1, tetA, qepA, macB, and vanR. This method employs ARG-specific dual probes combined with silver staining signal cascade amplification technology. A centrifuge-free concentration device has been developed that can directly enrich nucleic acid fragments from environmental samples, with traditional nucleic acid extraction and PCR amplification being eliminated. Using a dual probe combined with silver staining enhancement dual probe amplification technology, rapid high-throughput detection of low-concentration ARG is achieved. No specialized equipment is required, and on-site visual detection of ARG was realized. The detection method can detect target genes (sul1, tetA, qepA, macB, and vanR) within a concentration range of 0.411 µg mL⁻¹ to 55.9 µg mL⁻¹, with the lowest detectable target gene concentration being 0.411 µg mL⁻¹. This study marks the first integration of a centrifuge-free concentration device, specific dual probes, and silver staining signal cascade amplification technology to establish a rapid visual detection method for ARGs using a microarray. The developed detection method holds significant potential not only for the detection and monitoring of ARGs but also as a prototype for devising future detection strategies.