Real-time recombinase polymerase amplification assay for fast quantification of tetracycline resistance genes tetA in the surface water.

The tetracycline resistance gene tetA is a widely detected antibiotic resistance gene (ARG) posing significant ecological health risks in surface water. The development of rapid quantitative assays for tetA can substantially reduce both the time and economic costs associated with real-time monitoring of tetA transportation dynamics in the environment. In this study, a novel method for the quantification of tetracycline resistance gene tetA using real-time recombinase polymerase amplification was developed, which can complete the quantification of tetA within 20 minutes at a constant temperature of 39 ℃, achieving a detection limit of 50 copies/μL with 100 % sensitivity and specificity. Notably, this method facilitates the rapid quantification of tetA in surface water samples without the necessity for DNA extraction and purification, with a result deviation of less than 9.5 % compared to the conventional PCR method. This approach provides critical technical support for the on-site detection of tetracycline resistance genes in surface water.