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

Published on Jul 11, 2021in Foods4.35
· DOI :10.3390/FOODS10071605
Pingping Gao1
Estimated H-index: 1
,
Lihan Wang2
Estimated H-index: 2
+ 7 AuthorsYujun Jiang8
Estimated H-index: 8
Sources
Abstract
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 × 102 CFU/mL to 8.6 × 107 CFU/mL with the limit of detection (LOD) as low as 8.6 × 100 CFU/mL in pure culture. In actual samples, the visual LOD was 4.1 × 102 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.
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Abstract As a rapid and facile means for foodborne bacteria detection in situ, lateral flow immunoassay (LFA) still has intrinsic limitations in the construction of the existing sandwich LFA format, e.g. screening difficulties of paired antibodies (Abs), poor stability of Ab probe, etc. Here, combined the strong affinity of antibiotic with the superior specificity of antibody molecules, a novel and robust LFA based on a dual recognition strategy and magnetic separation was designed to achieve sp...
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