期刊
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
卷 414, 期 7, 页码 2333-2341出版社
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-022-03904-z
关键词
Single-tube; Diatomaceous earth; Nucleic acid amplification; Visual detection; Salmonella; Nucleic acid extraction
资金
- National Key Research and Development Programs of China [2018YFE0113300]
- Key Project of Shandong Provincial Natural Science Foundation [ZR2020KH030]
- National Natural Science Foundation of China [81801264]
Here, we developed an ultra-fast and visual single-tube nucleic acid detection approach that combines the self-settling characteristics of chitosan-functionalized diatomaceous earth (CDE) and accelerated PCR (AC-PCR). This method not only improves the sensitivity and specificity of detection, but also allows for the completion of the entire process in a short period of time, with results visible to the naked eyes.
Herein, we developed an ultra-fast and visual single-tube nucleic acid detection approach, which combined the advantages of self-settling characteristics of chitosan-functionalized diatomaceous earth (CDE) and accelerated PCR (AC-PCR). DNA was rapidly extracted by CDE within 3 min for the next nucleic acid amplification based on the nucleic acid attached on the chitosan in pH = 5.0. Under the action of gravity, the DNA-enriched CDE self-sediments to the bottom of the tube could be directly used for AC-PCR to achieve single-tube extraction and amplification. Our method detected Salmonella culture fluids with a detection limit of 1 CFU/mL, which was 100-fold more sensitive than conventional method that have not undergone nucleic acid enrichment. Furthermore, it also displayed high specificity and sensitivity for a variety of spiked samples. The entire process could be completed within 17 min in a single tube, and in particular, the result was visualized by the naked eyes. Overall, it is an all-in-one detection strategy without the requirement of redundant procedure, which greatly improved the detection efficiency, and saved the time and the cost. With these advantages, the approach will supply a promising tool in the field of point-of-care testing for Salmonella and other foodborne pathogens.
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