An Enhanced Lateral Flow Assay Based on Aptamer-Magnetic Separation and Multifold AuNPs for Ultrasensitive Detection of Salmonella Typhimurium in Milk

In this paper, a novel and ultrasensitive lateral flow assay (LFA) based on aptamer-magnetic separation, and multifold Au nanoparticles (AuNPs) was developed for visual detecting Salmonella enterica ser. Typhimurium (S. Typhimurium). The method realized magnetic enrichment and signal transduction via magnetic separation and achieved signal amplification through hybridizing AuNPs-capture probes and AuNPs-amplification probes to form multifold AuNPs. Two different thiolated single-strand DNA (ssDNA) on the AuNPs-capture probe played different roles. One was combined with the AuNPs-amplification probe on the conjugate pad to achieve enhanced signals. The other was connected to transduction ssDNA1 released by aptamer-magnetic capture of S. Typhimurium, and captured by the T-line, forming a positive signal. This method had an excellent linear relationship ranging from 8.6 x 10(2) CFU/mL to 8.6 x 10(7) CFU/mL with the limit of detection (LOD) as low as 8.6 x 10(0) CFU/mL in pure culture. In actual samples, the visual LOD was 4.1 x 10(2) CFU/mL, which did not carry out nucleic acid amplification and pre-enrichment, increasing three orders of magnitudes than unenhanced assays with single-dose AuNPs and no magnetic separation. Furthermore, the system showed high specificity, having no reaction with other nontarget strains. This visual signal amplificated system would be a potential platform for ultrasensitive monitoring S. Typhimurium in milk samples.

Automated summary

A novel lateral flow assay based on aptamer-magnetic separation and multifold Au nanoparticles was developed for visual detection of Salmonella enterica ser. Typhimurium. The method achieved high sensitivity and specificity without the need for nucleic acid amplification or pre-enrichment, significantly improving the detection limit compared to traditional assays.