A Novel Sensitive Recombinase-Aided Amplification Integrated Test Strip for in Milk via Dual Gene Probes.

is the main spoilage bacterium in milk, and its proliferation is one of the factors leading to the deterioration of the quality of raw milk. In this study, a rapid detection system for was developed based on recombinase-aided amplification combined with a test strip (RAA-TS), which contained a double test line (DTL) targeting the virulence gene of and the housekeeping gene of . Visual observation could detect (50 CFU/mL) and (250 CFU/mL) within 90 min, including sample pretreatment and RAA reaction and detection steps. No cross-reactions were observed with or other bacteria (n = 19). The quantitative detection limits (LOD) of and for in milk were 37 CFU/mL and 233 CFU/mL, respectively. Compared with polymerase chain reaction-agarose gel electrophoresis (PCR-AGE), the sensitivity of the developed RAA-TS-DTL system was increased by approximately four times. Furthermore, it could detect live in milk when combined with optimized sample pretreatment by propidium monoazide (PMAxx). Its consistency with the traditional culture method in the detection of spiked in milk samples (n = 25) was 100%. The developed RAA-TS-DTL had the advantages of high accuracy and short time consumption. Thus, it provides a new way or tool for the rapid screening or detection of in milk.