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.
Supplementary Information
The online version contains supplementary material available at 10.1007/s00216-022-03904-z.
Salmonella is one of the most common pathogens associated with foodborne illness in seafood. Herein, an accelerated strand exchange amplification (ASEA) requiring only a pair of primers and one polymerase...
Respiratory tract infections caused by Mycoplasma pneumoniae is a serious risk for child health. It has been difficult to prevent and control for a variety of reasons; therefore, timely diagnosis is particularly important for treatment of patients. At present, the rapid M pneumoniae test kits based on nucleic acid amplification have been commercialized and used as primary diagnostic tools for M pneumoniae infection, but current kits are time-consuming, which is difficult to meet the requirement for accurate and rapid diagnosis of M pneumoniae during epidemics. Rapid and accurate test kits are urgently required to diagnose M pneumoniae infection. In this article, we evaluated the performance of a novel nucleic acid detection kit (A) for M pneumoniae from feasibility and sensitivity, and compared it with kit B. Results showed this kit has the advantage of being rapid, sensitive, and specific, which meets the demands for the diagnosis of M pneumoniae infection in clinical settings.
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