Liver
cancer is a kind of high mortality cancer due to the difficulty
of early diagnosis. And according to the reports, the concentration
of reactive oxygen species (ROS) was higher in cancer cells than normal
cells. Therefore, developing an effective fluorescent probe for hepatoma-selective
imaging of hypochlorous acid (HOCl) which is one of the vital ROS
is of great importance for understanding the role of HOCl in liver
cancer pathogenesis. However, the cell-selective fluorescent probe
still remains a difficult task among current reports. Herein, a galactose-appended
naphthalimide (Gal-NPA) with p-aminophenylether
as a new receptor and galactose moiety as hepatoma targeting unit
was synthesized and employed to detect endogenous HOCl in living HepG2
cells. This probe was proved to possess good water solubility and
could respond specifically to HOCl. In addition, probe Gal-NPA could completely react to HOCl within 3 s meanwhile accompanied
by tremendous fluorescence enhancement. The quantitative linear range
between fluorescence intensities and the HOCl concentrations was 0
to 1 μM (detection limit = 0.46 nM). More importantly, fluorescence
confocal imaging experiments showed that probe Gal-NPA could discriminate hepatoma cells over other cancer cells and simultaneously
trace endogenous HOCl levels in living HepG2 cells. And we thus anticipate
that probe Gal-NPA has the potential application for
revealing the functions of HOCl in hepatoma cells.
Pertussis is a highly transmissible acute respiratory infection caused by the bacterial pathogen Bordetella pertussis. The purpose of this study was to develop a rapid, simple and sensitive diagnostic test for detecting this pathogen. Methods: Here we present a recombinase aided amplification (RAA) assay incorporating competitive internal amplification control (IAC) to detect Bordetella pertussis using the DNA obtained by boiling. This assay was performed in a single closed tube at 39 C within 30 min. A total of 115 clinical samples suspected of pertussis were collected and tested by the internally controlled RAA assay using both extracted DNA with the commercial kit and the DNA obtained by boiling. For comparison, the real-time PCR (RT-PCR) was also performed with DNA extraction in parallel. Results: The sensitivity of the internally controlled RAA assay was 10 1 copies or 10 CFU/ml per reaction in detecting plasmid DNA or B. pertussis strain. The optimum concentration of the IAC plasmid was determined to be 100 copies, and the introduction of IAC effectively reduced the occurrence of false negatives. Compared to the RT-PCR, RAA results with DNA extraction obtained 100% sensitivity and specificity, and the RAA results with heat-treated DNA showed 85.96% sensitivity and 100% specificity. Conclusion: With the advantages of 45 min turnaround time and simple steps of DNA purification, this assay could become a useful diagnostic tool for Bordetella pertussis detection and is potentially suitable for point-of-care identification to guide prompt clinical treatment.
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