1. Hybrid RCA-DLS assay combined with aPCR for sensitive Salmonella enteritidis detection.
- Author
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Xie G, Zhan Z, Ye Y, Zhou B, Tong P, Aguilar ZP, and Xu H
- Subjects
- DNA, Bacterial analysis, DNA, Bacterial genetics, Dynamic Light Scattering, Gold, Limit of Detection, Polymerase Chain Reaction methods, Metal Nanoparticles, Salmonella enteritidis genetics
- Abstract
Salmonella infection could come from eating contaminated meat or raw eggs, and drinking milk or water contaminated by Salmonella enteritidis. Therefore, it is necessary to explore a fast and easy method for the detection of S. enteritidis in these diverse samples. For this purpose, a novel particle size sensing tool was designed for ultrasensitive and accurate S. enteritidis detection. This assay consisted of rolling circle amplification (RCA) with dynamic light scattering (DLS) using gold nanoparticles (AuNPs) modified with DNA probe as DNA-AuNPs as the capture surface into a hybrid RCA-DLS assay combined with asymmetric polymerase chain reaction (aPCR) and subsequent detection. Under optimal experimental conditions, the novel hybrid RCA-DLS assay combined with aPCR for S. enteritidis reached a limit of detection (LOD) as low as 3 × 10
0 CFU/mL in pure culture. In spiked milk samples, the LOD was 2.0 × 100 CFU/mL without pre-enriched bacteria. The total time of RCA-DLS assay was about 6 h which including genomic DNA extraction, aPCR, RCA and DLS determination. The hybrid RCA-DLS assay combined with aPCR holds promise in the specific and sensitive S. enteritidis detection., (Copyright © 2022 Elsevier Inc. All rights reserved.)- Published
- 2022
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