In this work, two different deposition methods of 3-aminopropyltriethoxysilane (APTES) on glass slides were compared in order to study the adhesion effect of cervical exfoliated cells on smear slides. Glass slides were modified by vapor-phase deposition (V-D) and liquid-phase deposition (L-D), respectively. The topographic images and amine density of the modified slides were investigated by using atomic force microscopy, UV-vis spectroscopy and X-ray photoelectron spectroscopy. The numbers of cells adhered on the slides functionalized by V-D and L-D were counted and compared under the microscope. The data showed significant differences between the two methods (t-test: P < 0.05). The results presented here have made it theoretically possible to produce amine slides by V-D method for the ThinPrep cytologic test.
Pathogen infections present a considerable
threat to global health
owing to the high morbidity and mortality, and usually multiple pathogens
coexist in food and the environment. Consequently, it is in urgent
need to develop some multiplexed and sensitive approaches for pathogen
detection. Here, we presented a novel strategy using mass tag-mediated
surface engineering for simultaneous detection of multiple bacteria
by matrix-assisted laser desorption/ionization-time of flight mass
spectrometry (MALDI-TOF MS). Following aptamer binding, primer amplification,
and DNA hybridization, bacteria were specifically labeled by their
corresponding mass tags, which could be released and ionized after
laser irradiation. This strategy converted the detection of bacteria
to the analysis of mass tags, allowing simultaneous detection of multiple
bacteria and avoiding the dependence of microbial mass spectra databases.
In addition, this approach applied two rolling circle amplification
(RCA) reactions to improve both the capture efficiency and detection
sensitivity of the target bacteria. The specificity and the real sample
detection were evaluated, and the results demonstrated a potential
application of this approach in milk safety monitoring.
A portable microfluidic biosensor was developed for the detection of E. coli O157:H7 using finger actuation. The chip was assembled with three functional zones, immunomagnetic separation, nucleic acid extraction and purification, and signal detection. First, antibody-modified magnetic nanoparticles (MNPs) were used to separate the target bacteria from the sample. The captured bacteria were then lysed and silica-coated MNPs were used to absorb DNA, followed by washing and eluting to obtain purified DNA. The obtained DNA was subjected to amplification and fluorescence detection based on the recombinase polymerase amplification-clustered regularly interspaced short palindromic repeat-associated protein/Cas12a reaction. The fluorescence images were collected and analyzed using a smartphone app under a 3D-printed detection device. It could quantitatively detect E. coli O157:H7 from 10 2 to 10 8 CFU/mL in 2.5 h with a limit of detection (LOD) of 10 CFU/mL. The recovery rate ranged from 104 to 120%. Overall, the biosensor realizes "sample-in and answer-out" assay for E. coli O157:H7 and eliminates the need for external pumps and skilled personnel.
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