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Lateral flow fluorescent immunoassay based on isothermal amplification for rapid quantitative detection of Salmonella spp. | LitMetric

Lateral flow fluorescent immunoassay based on isothermal amplification for rapid quantitative detection of Salmonella spp.

Analyst

State Key Laboratory of Bioelectronics, Jiangsu Key Laboratory for Biomaterials and Devices, School of Biological Science and Medical Engineering and Collaborative Innovation Center of Suzhou Nano Science and Technology, Southeast University, Nanjing 210096, P. R. China.

Published: March 2020

AI Article Synopsis

  • Salmonella spp. are harmful germs that pose significant public health risks, and a new rapid test called SEA-LFIA has been developed for their detection without needing complex equipment.
  • This method uses a specific enzyme and fluorescent technology to identify Salmonella with a high level of accuracy, tested against numerous strains.
  • The SEA-LFIA is quick (taking about 30 minutes), cost-effective, and can detect low levels of Salmonella in various samples, making it a promising tool for on-site diagnostics in the field.

Article Abstract

Salmonella spp. are zoonotic pathogens of substantial public health concern. To enable detection in the field or under instrument-free conditions, we developed a rapid and robust lateral flow fluorescent immunoassay based on strand exchange amplification (SEA-LFIA) for the quantitative detection of Salmonella spp. As far as we know, this work is the first report regarding the use of Bst DNA polymerase-assisted SEA for fluorescence sensing to detect Salmonella spp. The SEA method was further confirmed by enzymatic digestion and Sanger dideoxy sequencing. The specificity of SEA-LFIA assay was verified by 89 Salmonella strains (18 Salmonella reference strains and 71 clinical isolates) and 15 non-Salmonella reference strains (different genera). The sensitivity of SEA-LFIA assay was 6 × 10 CFU mL of Salmonella pure culture or 3 × 10 CFU 25 g of artificially spiked raw chicken meat. Using this assay, it was found that 37 (16%) of the 236 samples collected were positive, which was consistent with the results of conventional PCR. The cutoff value is 15 and SEA-LFIA assay only takes ∼30 min without high equipment and reagent cost. In addition, the proposed strategy can be easily extended by redesigning the corresponding amplification primers to detect target analytes. In conclusion, the optimized SEA-LFIA assay is an efficient and specific method for the detection of Salmonella spp., and can potentially serve as a new on-site diagnostic tool in life sciences.

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Source
http://dx.doi.org/10.1039/c9an02011jDOI Listing

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