ABSTRACT
The electromagnetic eddy current non-destructive testing system enables the non-destructive analysis of surface defect information on tested materials. Based on the principles of eddy current detection, this paper presents a digital eddy current detection method using high-speed sampling based on STM32. A differential eddy current coil is used as the detection probe, and the combination of a differential bridge and a differential amplifier circuit helps to reduce common-mode noise interference. The detection signal is collected via an STM32-based acquisition circuit and transmitted to the host computer through Ethernet for digital demodulation processing. The host computer performs operations such as smoothing averaging, sinusoidal fitting, and outlier removal to extract the amplitude and phase of the detection signal. The system also visually displays the condition of the tested object's surface in real time through graphical visualization. Testing showed that this system can operate at frequencies up to 8.84 MHz and clearly identify defects as narrow as 1 mm on the surface of the tested steel plate.
ABSTRACT
Single-nucleotide mutations (SNMs) in the bacterial genome may cause antibiotic resistance. The visualization of SNMs can indicate antibiotic resistance phenotypes at the single-cell level but remains challenging. Herein, we proposed an in situ allele-specific isothermal amplification proceeded inside cells, allowing us to image bacterial genes with single-nucleotide resolution. The primer for loop-mediated isothermal amplification (LAMP) was designed with artificial mismatch bases to serve as an allele-specific probe, endowing LAMP to specifically amplify genes with SNMs. Due to the high amplification efficiency of LAMP, the method termed AlleLAMP can generate high gain for imaging SNMs and precisely quantify mutated quinolone-resistant Salmonella in bacterial mixture. We utilized AlleLAMP to survey the selection of antibiotic resistance under the preservative stress and found that the mutant quinolone-resistant strain owned a survival advantage over the wild-type quinolone-sensitive strain under the stress of preservatives. AlleLAMP can serve as a single-cell tool for analyzing the relationship between bacterial genotype and phenotype.
Subject(s)
Nucleotides , Quinolones , Genotype , Alleles , MutationABSTRACT
Staphylococcus aureus (S. aureus) causes serious food-borne diseases, and tools able to directly profile intact S. aureus would greatly facilitate food safety and public health. Herein, we proposed a biosensing platform for culture-independent and separation-free profiling S. aureus, thus allow us to directly detect intact S. aureus in complex samples. The binding protection effect of aptamer-cell complex was introduced to construct the aptasensor, and it allowed to eliminate the optimization of aptamer probe sequences. The proposed aptasensor, terms enzymatic cleavage aptasensor could achieve a sensitive (a detection limit of 64 CFU/mL) and broad-concentration quantification (dynamic range 102-107 CFU/mL) of S. aureus. Furthermore, it could specifically identify intact S. aureus in complex samples, and the quantifying of S. aureus was achieved in tap water, milk and porker with high precision. Therefore, enzymatic cleavage aptasensor could be a good candidate for on-site biosensing platform of S. aureus, as well as other pathogens by replacing the aptamer sequences.
Subject(s)
Aptamers, Nucleotide , Biosensing Techniques , Staphylococcal Infections , Animals , Limit of Detection , Staphylococcus aureus , WaterABSTRACT
Excessive use of antibiotics in aquatic products is a serious problem for food safety and human health, and on-site detection of antibiotics is highly demanded. Herein, we proposed multivalence aptamer probes, allowing sensitive, label-free, and homogeneous detection of antibiotics in different aquatic products. Compared to commonly used aptamers, multivalence aptamer probes can provide multiple binding sites and a higher affinity for target molecules, and the iterative binding on different binding sites contributes to an amplified recognition effect, sharply increasing the response and sensitivity of aptamer probes. The 2-valence aptamer probes conferred a limit of detection of 0.097 nM for kanamycin detection, where it is estimated that their sensitivity is enhanced 12 times compared to 1-valence aptamer probes. Meanwhile, multivalence aptamer probes allowed us to specifically identify kanamycin among other antibiotics. It could detect kanamycin residual in aquatic products including river eel and puffer fish, as well as tap water with high precision. A multivalence design strategy of aptamer probes would significantly improve the detection performance of aptamers, facilitating the translation of aptamer for food safety control.