Ullah N.Suchanta N.Pimpitak U.Santanirand P.Am-In N.Chaichanawongsaroj N.Mahidol University2024-11-062024-11-062024-10-01Antibiotics Vol.13 No.10 (2024)https://repository.li.mahidol.ac.th/handle/20.500.14594/101907Background/Objectives: The emergence of the mobilized colistin resistance 1 (mcr-1) gene, which causes colistin resistance, is a serious concern in animal husbandry, particularly in pigs. Although antibiotic regulations in many countries have prohibited the use of colistin in livestock, the persistence and dissemination of this plasmid-mediated gene require effective and rapid monitoring. Therefore, a rapid, sensitive, and specific method combining recombinase polymerase amplification (RPA) with an in-house lateral flow assay (LFA) for the mcr-1 gene detection was developed. Methods: The colistin agar test and broth microdilution were employed to screen 152 E. coli isolates from pig fecal samples of five antibiotic-used farms. The established RPA-in-house LFA was validated with PCR for mcr-1 gene detection. Results: The RPA-in-house LFA was completed within 35 min (20 min of amplification and 5–15 min on LFA detection) at 37 °C. The sensitivity, specificity, and accuracy were entirely 100% in concordance with PCR results. No cross-reactivity was detected with seven common pathogenic bacteria or other mcr gene variants. Conclusions: Therefore, the in-house RPA-LFA serves as a point-of-care testing tool that is rapid, simple, and portable, facilitating effective surveillance of colistin resistance in both veterinary and clinical settings, thereby enhancing health outcomes.Pharmacology, Toxicology and PharmaceuticsBiochemistry, Genetics and Molecular BiologyMedicineImmunology and MicrobiologyArticleSCOPUS10.3390/antibiotics131009842-s2.0-8520764023120796382