A ratiometric SERS aptasensor based on catalytic hairpin self-assembly mediated cyclic signal amplification strategy for the reliable determination of E. coli O157:H7.

A ratiometric SERS aptasensor based on catalytic hairpin self-assembly (CHA) mediated cyclic signal amplification strategy was developed for the rapid and reliable determination of Escherichia coli O157:H7. The recognition probe was synthesized by modifying magnetic beads with blocked aptamers, and the SERS probe was constructed by functionalizing gold nanoparticles (Au NPs) with hairpin structured DNA and 4-mercaptobenzonitrile (4-MBN). The recognition probe captured E. coli O157:H7 specifically and released the blocker DNA, which activated the CHA reaction on the SERS probe and turned on the SERS signal of 6-carboxyl-x-rhodamine (ROX). Meanwhile, 4-MBN was used as an internal reference to calibrate the matrix interference. Thus, sensitive and reliable determination and quantification of E. coli O157:H7 was established using the ratio of the SERS signal intensities of ROX to 4-MBN. This aptasensor enabled detection of 2.44 × 102 CFU/mL of E. coli O157:H7 in approximately 3 h without pre-culture and DNA extraction. In addition, good reliability and excellent reproducibility were observed for the determination of E. coli O157:H7 in spiked water and milk samples. This study offered a new solution for the design of rapid, sensitive, and reliable SERS aptasensors.

Recent Progress on Toxicity and Detection Methods of Polychlorinated Biphenyls in Environment and Foodstuffs.

Polychlorinated biphenyls (PCBs), a class of synthetic organochlorine chemicals, were broadly employed in industrial and commercial applications in the last century due to their good thermal and chemical stability. However, PCBs have a great influence on both individual organism and the entire ecosystem. It has been proven that PCBs pose potential risks to human health with neurotoxicity, carcinogenicity, reproductive toxicity, immunotoxicity, hepatotoxicity, and cardiovascular toxicity. Moreover, PCBs exhibit the long-range transport effect on the global scale and bio-enrichment effect along the food chains. This review mainly encompasses recent progress on the toxicity and detection techniques of PCBs in environment and foodstuffs. First of all, we highlighted the latest improvements and achievements of the classification, source, distribution, and toxicity of PCBs. Then, comprehensive summaries of the current technologies for sample preparation (e.g., SPE, DSPE, SPME and SBSE) and analytical determination (e.g., GC-ECD, GC-MS, GC-HRMS, HPLC-MS/MS and sensing technologies) were given. In the end, the shortcomings and prospects of the pretreatment methods for PCBs analysis as well as the future opportunities and challenges are tentatively discussed.

Reliable and Rapid Detection and Quantification of Enrofloxacin Using a Ratiometric SERS Aptasensor.

Reliable detection and quantification of antibiotic residues in food using surface-enhanced Raman spectroscopy remain challenging, since the intensities of SERS signals are vulnerable to matrix and experimental factors. In this work, a ratiometric SERS aptasensor using 6-Carboxyl-X-Rhodamine (ROX)-labeled aptamers and 4-mercaptobenzonitrile (4-MBN)-functionalized gold nanoparticles (Au NPs) as SERS probes was established for the reliable and rapid detection and quantification of enrofloxacin. In the presence of enrofloxacin, the conformational transform of aptamers took place, and the distance between ROX and Au NP increased, which resulted in a decrease in the SERS signal intensity of ROX. Meanwhile, the intensity of the SERS signal of 4-MBN was used as an internal standard. Reliable determination of enrofloxacin was realized using the ratio of the SERS signal intensities of ROX to 4-MBN. Under optimal conditions, the developed ratiometric SERS aptasensor provided a wide linear range from 5 nM to 1 µM, with a correlation coefficient (R2) of 0.98 and a limit of detection (LOD) of 0.12 nM (0.043 ppb). In addition, the developed ratiometric SERS aptasensor was successfully applied for the determination of enrofloxacin in fish and chicken meat, with recovery values of 93.6-112.0%. Therefore, the established ratiometric SERS aptasensor is sensitive, reliable, time-efficient, and has the potential to be applied in the on-site detection of enrofloxacin in complex matrices.

Rapid and reliable detection and quantification of organophosphorus pesticides using SERS combined with dispersive liquid-liquid microextraction.

Rapid and reliable detection and quantification of pesticide residues in complex matrices by surface enhanced Raman spectroscopy (SERS) remain challenging due to the low level of target molecules and the interference of nontarget components. In this study, SERS was combined with dispersive liquid-liquid microextraction (DLLME) to develop a rapid and reliable method for the detection of organophosphorus pesticides (OPPs). In this method, DLLME was used to extract and enrich two representative OPPs (triazophos and parathion-methyl) from a liquid sample, and a portable Raman spectrometer was used to analyze the separated sediment using homemade gold nanoparticles colloids as enhancing substrates. The results showed that the developed method displayed good sensitivity and stability for the detection and quantification of triazophos and parathion-methyl with R2 ≥ 0.98. The calculated limits of detection (LODs) in the simultaneous detection of triazophos and parathion-methyl were 2.17 × 10-9 M (0.679 ppb) and 2.28 × 10-8 M (5.998 ppb), and the calculated limits of quantification (LOQs) were 7.23 × 10-9 M (2.26 ppb) and 7.62 × 10-8 M (19.098 ppb), respectively. Furthermore, the developed SERS method was successfully applied to the detection of triazophos and parathion-methyl in apple juice with recoveries between 78.07% and 110.87% and relative standard deviations (RSDs) ≤ 2.06%. Therefore, the developed DLLME facilitated liquid SERS method exhibited good sensitivity and stability for the rapid detection and quantification of OPPs and had the potential to be applied to the rapid detection of OPPs in complex matrices.

Label-free quantitative proteomic analysis of the mechanism of salt stress promoting selenium enrichment in Lactobacillus rhamnosus.

Lactobacillus rhamnosus can metabolize selenite into organic selenium and Se0. In this paper, label-free quantitative proteomics was applied to explore the mechanism of salt stress promoting selenium enrichment of L.rhamnosus. 397 proteins were up-regulated and 147 proteins were down-regulated of selenium-enriched L.rhamnosus under salt stress. The differentially expressed proteins (DEPs) were mainly involved in metabolism, membrane transport and genetic information processing. The results of quantitative real-time PCR showed that gene opuA, metN, trxR and ldh of Se-enriched L.rhamnosus with salt stress were significantly up-regulated. However, the expression levels of gene luxS, groEL, dnaK and pgk were down-regulated. It was indicated that L.rhamnosus promoted part of amino acids combining with selenium into selenoamino acids with salt stress. Secondly, sodium chloride stimulated the expression of key enzymes involved in metabolism to provide energy for the process of Se-enrichment. In addition, NaCl induced the expression of enzymes and genes involved in the synthesis of selenoproteins. SIGNIFICANCE: It was indicated that L.rhamnosus promoted part of amino acids combining with selenium into selenoamino acids with salt stress. Secondly, sodium chloride stimulated the expression of key enzymes involved in metabolism to provide energy for the process of Se-enrichment. In addition, NaCl induced the expression of enzymes and genes involved in the synthesis of selenoproteins.

Ultrasensitive CRISPR/Cas12a-Driven SERS Biosensor for On-Site Nucleic Acid Detection and Its Application to Milk Authenticity Testing.

An ultrasensitive surface-enhanced Raman scattering (SERS) biosensor driven by CRISPR/Cas12a was proposed for on-site nucleic acid detection. We tactfully modified single-strand DNA (ssDNA) with a target-responsive Prussian blue (PB) nanolabel to form a probe and fastened it in the microplate. Attributed to the specific base pairing and highly efficient trans-cleavage ability of the CRISPR/Cas12a effector, precise target DNA recognition and signal amplification can be achieved, respectively. In the presence of target DNA, trans-cleavage towards the probe was activated, leading to the release of a certain number of PB nanoparticles (NPs). Then, these free PB NPs would be removed. Under alkali treatment, the breakdown of the remaining PB NPs in the microplate was triggered, producing massive ferricyanide anions (Fe(CN)64-), which could exhibit a unique characteristic Raman peak that was located in the "biological Raman-silent region". By mixing the alkali-treated solution with the SERS substrate, Au@Ag core-shell NP, the concentration of the target DNA was finally exhibited as SERS signals with undisturbed background, which can be detected by a portable Raman spectrometer. Importantly, this strategy could display an ultralow detection limit of 224 aM for target DNA. Furthermore, by targeting cow milk as the adulterated ingredient in goat milk, the proposed biosensor was successfully applied to milk authenticity detection.

Highly sensitive multiplex detection of foodborne pathogens using a SERS immunosensor combined with novel covalent organic frameworks based biologic interference-free Raman tags.

Rapid and reliable multiplex detection of foodborne pathogens is in great demand for ensuring food safety and preventing foodborne diseases. In this study, we developed a highly sensitive SERS immunosensor for the simultaneous detection of multiple foodborne pathogens. Novel biologic interference-free Raman tags synthesized by using the covalent organic frameworks (COF) TBDP as nanocontainer to load biologic interference-free Raman reporters and specific antibodies for interested targets were used to convert and amplify signals of foodborne pathogens. In addition, lectin functionalized magnetic nanoparticles (MNPs@Con A) which could efficiently bind to the carbohydrate constituents on the surface of pathogens were prepared to capture and isolate multiple pathogens simultaneously. The recognition of the target foodborne pathogen impels the generation of sandwich-like composites of MNPs@Con A/pathogen/TBDP@Raman tags, and these composites could be quickly separated from the sample matrix with the assistance of an external magnet. Besides, a mass of Raman reporters was released by eluting the collected MNPs@Con A/pathogen/TBDP@Raman tags composites. Combined with a portable Raman system, characteristic Raman signals (2271 and 2113 cm-1) of the occupied reporters located at the biologic interference-free region were observed and used for the simultaneous detection of two different foodborne pathogenic strains. And an equal limit of detection of 101 CFU/mL was achieved for each strain. This strategy provides new insight into the application of SERS in the detection of pathogenic bacteria.

Recent advances of sensing strategies for the detection of ß-glucuronidase activity.

ß-Glucuronidase (ß-GLU), a kind of hydrolase, is widely distributed in mammalian tissues, body fluids, and microbiota. Abnormal changes of ß-GLU activity are often correlated with the occurrence of diseases and deterioration of water quality. Therefore, detection of ß-GLU activity is of great significance in biomedicine and environmental health such as cancer diagnosis and water monitoring. However, the conventional ß-GLU activity assay suffers from the limitations of low sensitivity, poor accuracy, and complex procedure. With the development of analytical chemistry, many advances have been made in the detection of ß-GLU activity in recent years. The sensors for ß-GLU activity detection which have the advantages of rapid and reliable detection have been attracting increased attentions. In this paper, the principles, performances, and limitations of these ß-GLU sensors, including colorimetric sensing, fluorescent sensing, electrochemical sensing for the determination of ß-GLU activity, have been summarized and discussed. Moreover, the challenges and research trends of ß-GLU activity assay are proposed.

Recent advances in dual recognition based surface enhanced Raman scattering for pathogenic bacteria detection: A review.

Rapid and reliable detection of pathogenic bacteria at the early stage represents a highly topical research area for food safety and public health. Although culture based method is the gold standard method for bacteria detection, recent techniques have promoted the development of alternative methods, such as surface enhanced Raman scattering (SERS). SERS provides additional advantages of high speed, simultaneous detection and characterization, multiplex analysis, and comparatively low cost. However, conventional SERS methods for bacteria detection are facing limitations of low sensitivity, susceptible to matrix interference, and poor accuracy. In recent years, specific detection of pathogenic bacteria with dual recognition based SERS methods has attracted increasing attentions. These methods include two steps recognition of target bacteria, and integrate the functions of target separation and detection. Considering their merits of excellent specificity, ultrahigh sensitivity, multiplex detection capability, and potential for on-site applications, these methods are promising alternatives for rapid and reliable detection of pathogenic bacteria. Herein, this review aims to summarize the recent advances in dual recognition based SERS methods for specific detection of pathogenic bacteria. Their advantages and limitations are discussed, and further perspectives are tentatively given. This review provides new insights into the application of SERS as a reliable tool for pathogenic bacteria detection.