Phenotypic and genotypic perspectives on detection methods for bacterial antimicrobial resistance in a One Health context: research progress and prospects.

The widespread spread of bacterial antimicrobial resistance (AMR) and multidrug-resistant bacteria poses a significant threat to global public health. Traditional methods for detecting bacterial AMR are simple, reproducible, and intuitive, requiring long time incubation and high labor intensity. To quickly identify and detect bacterial AMR is urgent for clinical treatment to reduce mortality rate, and many new methods and technologies were required to be developed. This review summarizes the current phenotypic and genotypic detection methods for bacterial AMR. Phenotypic detection methods mainly include antimicrobial susceptibility tests, while genotypic detection methods have higher sensitivity and specificity and can detect known or even unknown drug resistance genes. However, most of the current tests are either genotypic or phenotypic and rarely combined. Combining the advantages of phenotypic and genotypic methods, combined with the joint application of multiple rapid detection methods may be the trend for future AMR testing. Driven by rapid diagnostic technology, big data analysis, and artificial intelligence, detection methods of bacterial AMR are expected to constantly develop and innovate. Adopting rational detection methods and scientific data analysis can better address the challenges of bacterial AMR and ensure human health and social well-being.

Formaldehyde contamination in seafood industry: an update on detection methods and legislations.

Seafood is abundant in high-quality protein, healthy fats (n-3 and n-6 PUFAs), minerals (calcium, magnesium, copper, selenium, and so on), and vitamin D. Functional compounds in seafood can protect against lifestyle-related diseases. Having had all the merits mentioned, it is also a highly putrefiable food item. To maintain quality and prolong seafood's shelf life, various chemicals have been added, including nitrite, sulfur dioxide, and formaldehyde. In this review, we summarize the uses, the incidence of added formalin contamination, and the approved limit for seafood products. Additionally, worldwide regulations or standards for the use of formalin in seafood products, as well as recent changes relevant to new methods, are highlighted. Although strict limits and regulations have been placed on the utilization of formaldehyde for seafood preservation, there are few incidences reported of formalin/formaldehyde detection in seafood products around Asian countries. In this context, various qualitative and quantitative detection methods for formaldehyde have been developed to ensure the presence of formaldehyde within acceptable limits. Besides this, different rules and regulations have been forced by each country to control formaldehyde incidence. Although it is not an issue of formaldehyde incidence in European countries, strict regulations are implemented and followed.

Recent progress of methods for cuproptosis detection.

Varying from other identified cell death pathways, cuproptosis is a new type of regulated cell death characterized by excess Cu ions, abnormal aggregation of lipoylated proteins in TCA cycle, loss of Fe-S cluster proteins, upregulation of HSP70, leading to proteotoxic and oxidative stress. Cuproptosis is highly concerned by scientific community and as the field of cuproptosis further develops, remarkable progress has been made in the verification and mechanism of cuproptosis, and methods used to detect cuproptosis have been continuously improved. According to the characteristic changes of cuproptosis, techniques based on cell death verification, Cu content, morphology, molecular biology of protein levels of cuproptosis-related molecules and biochemical pathways of cuproptosis-related enzyme activity and metabolites of oxidative stress, lipoic acid, TCA cycle, Fe-S cluster proteins, oxidative phosphorylation, cell respiration intensity have been subject to cuproptosis verification and research. In order to further deepen the understanding of detecting cuproptosis, the principle and application of common cuproptosis detection methods are reviewed and categorized in cellular phenomena and molecular mechanism in terms of cell death, Cu content, morphology, molecular biology, biochemical pathways with a flow chart. All the indicating results have been displayed in response to the markers of cuproptosis, their advantages and limitations are summaried, and comparison of cuproptosis and ferroptosis detection is performed in this study. Our collection of methods for cuproptosis detection will provide a great basis for cuproptosis verification and research in the future.

Recent Development in Detection and Control of Psychrotrophic Bacteria in Dairy Production: Ensuring Milk Quality.

Milk is an ideal environment for the growth of microorganisms, especially psychrotrophic bacteria, which can survive under cold conditions and produce heat-resistant enzymes. Psychrotrophic bacteria create the great problem of spoiling milk quality and safety. Several ways that milk might get contaminated by psychrotrophic bacteria include animal health, cowshed hygiene, water quality, feeding strategy, as well as milk collection, processing, etc. Maintaining the quality of raw milk is critically essential in dairy processing, and the dairy sector is still affected by the premature milk deterioration of market-processed products. This review focused on the recent detection and control strategies of psychrotrophic bacteria and emphasizes the significance of advanced sensing methods for early detection. It highlights the ongoing challenges in the dairy industry caused by these microorganisms and discusses future perspectives in enhancing milk quality through innovative rapid detection methods and stringent processing controls. This review advocates for a shift towards more sophisticated on-farm detection technologies and improved control practices to prevent spoilage and economic losses in the dairy sector.

Occurrence of Uncultured Legionella spp. in Treated Wastewater Effluent and Its Impact on Human Health (SCA.Re.S Project).

Wastewater treatment plants (WWTPs) provide optimal conditions for the environmental spread of Legionella. As part of the Evaluation of Sanitary Risk Related to the Discharge of Wastewater to the Ground (SCA.Re.S) project, this study was conducted to evaluate the presence of Legionella in WWTP effluent and in groundwater samples collected from two wells located downstream from the plant. The samples were analyzed to determine the concentrations of Legionella spp using the standard culture-based method and molecular techniques, followed by genomic sequencing analysis. Legionella was detected only with the molecular methods (except in one sample of effluent positive for L. pneumophila serogroup 6), which showed viable Legionella pneumophila and L. non-pneumophila through the use of free DNA removal solution in both the effluent and groundwater, with concentrations that progressively decreased downstream from the plant. Viable L. pneumophila appeared to be slightly more concentrated in warm months. However, no significant differences (p ≥ 0.05) in concentrations between cold and warm months were observed. A genotypic analysis characterized the species present in the samples and found that uncultured Legionella spp, as yet undefined, constituted the prevalent species in all the samples (range 77.15-83.17%). WWTPs play an important role in the hygienic and sanitary quality of groundwater for different uses. The application of Legionella control systems during the purification of effluents is warranted to prevent possible outbreaks of legionellosis.

Development of Biolayer Interferometry (BLI)-Based Double-Stranded RNA Detection Method with Application in mRNA-Based Therapeutics and Vaccines.

Background: In vitro-transcribed (IVT) mRNA has been established as a promising platform for therapeutics and vaccine development. Double-stranded RNA (dsRNA) is a major impurity of IVT mRNA and can trigger unfavored immune responses, potentially causing adverse events in patients. Existing dsRNA detection and quantitation methods, such as gel electrophoresis, ELISA, or homogeneous time-resolved fluorescence (HTRF), have low sensitivity or are time-consuming. A recently published lateral flow immunoassay (LFSA) was shown to be fast, but it lacks the sensitivity for dsRNA with uridine modifications. Methods: In this study, we provided a possible explanation for the reduced sensitivity of existing quantitation methods for dsRNA with modified uridines by characterizing the binding affinities of commonly used anti-dsRNA antibodies. Then, a rapid and sensitive biolayer interferometry (BLI) dsRNA detection assay utilizing Flock House Virus (FHV) B2 protein was developed to overcome the challenges in dsRNA detection and the reduced sensitivity. Results: This assay allows the detection of dsRNA with different uridine modifications (ψ, m1ψ, 5 moU) with similar sensitivity as dsRNA without modification. Furthermore, we demonstrated this method can be used to quantify both short and long dsRNA, as well as hairpin-structured dsRNA, providing a more comprehensive detection for dsRNA impurities. Moreover, we applied this assay to monitor dsRNA removal through a purification process. Conclusions: Taken together, this BLI method could enable real-time monitoring of impurities in IVT mRNA production to prevent immunogenicity stemming from dsRNA.

Linker-specific monoclonal antibodies present a simple and reliable detection method for scFv-based CAR NK cells.

Chimeric antigen receptor (CAR) T cell therapies have demonstrated significant successes in treating cancer. Currently, there are six approved CAR T cell products available on the market that target different malignancies of the B cell lineage. However, to overcome the limitations of CAR T cell therapies, other immune cells are being investigated for CAR-based cell therapies. CAR natural killer (NK) cells can be applied as allogeneic cell therapy, providing an economical, safe, and efficient alternative to autologous CAR T cells. To improve CAR research and future in-patient monitoring of cell therapeutics, a simple, reliable, and versatile CAR detection reagent is crucial. As most existing CARs contain a single-chain variable fragment (scFv) with either a Whitlow or a G4S linker site, linker-specific monoclonal antibodies (mAbs) can detect a broad range of CARs. This study demonstrates that these linker-specific mAbs can detect different CAR NK cells in vitro, spiked in whole blood, and within patient-derived tumor spheroids with high specificity and sensitivity, providing an effective and almost universal alternative for scFv-based CAR detection. Additionally, we confirm that linker-specific antibodies can be used for functional testing and enrichment of CAR NK cells, thereby providing a useful research tool to fast-track the development of novel CAR-based therapies.

On the Utility of Indirect Methods for Detecting Faking.

Indirect indices for faking detection in questionnaires make use of a respondent's deviant or unlikely response pattern over the course of the questionnaire to identify them as a faker. Compared with established direct faking indices (i.e., lying and social desirability scales), indirect indices have at least two advantages: First, they cannot be detected by the test taker. Second, their usage does not require changes to the questionnaire. In the last decades, several such indirect indices have been proposed. However, at present, the researcher's choice between different indirect faking detection indices is guided by relatively little information, especially if conceptually different indices are to be used together. Thus, we examined and compared how well indices of a representative selection of 12 conceptionally different indirect indices perform and how well they perform individually and jointly compared with an established direct faking measure or validity scale. We found that, first, the score on the agreement factor of the Likert-type item response process tree model, the proportion of desirable scale endpoint responses, and the covariance index were the best-performing indirect indices. Second, using indirect indices in combination resulted in comparable and in some cases even better detection rates than when using direct faking measures. Third, some effective indirect indices were only minimally correlated with substantive scales and could therefore be used to partial faking variance from response sets without losing substance. We, therefore, encourage researchers to use indirect indices instead of direct faking measures when they aim to detect faking in their data.

Critical reassessment of microplastic abundances in the marine environment.

Microplastics (MPs) pose a growing concern in the marine environment, but their global prevalence remains largely unknown due to the absence of precise and standardized detection methods. This review critically evaluates existing techniques for quantifying MP abundances in marine field studies, addressing inaccuracies resulting from the exclusion of particle sizes, polymer types, or limitations in identification methods. These traced inaccuracies were considered to recalculate MP abundances for particle sizes from 10 to 5000 µm, providing the first corrected global overview of MP distribution that enables quality assessment and reliable comparisons between adjusted data. The recalculations indicate that MP abundances are up to 15 times higher in marine waters (average (1.5 ± 36.2) × 105 items m-3) and up to 11 times higher in the marine sediments (average (2.7 ± 117.9) × 105 items kg-1) than previously reported in the literature. The Australasian Mediterranean Sea (average (1.2 ± 10.6) × 106 items m-3) and the North Atlantic (average (2.1 ± 37.6) × 105 items kg-1) emerged as the most polluted regions in marine waters and sediments, respectively, with primary contributors being the coasts of Southeast Asia and East America. This review demonstrates that previous field studies, global estimates, and models have significantly underestimated MP levels in marine environments in many cases, which could result in misinterpretations of both local and global pollution levels. This work highlights the critical need for precise handling of microplastic samples and urges future researchers to adopt standardized protocols for MP analysis to avoid inaccurate and misleading outcomes.

Advances in understanding the molecular mechanisms of borderline ovarian tumors.

Borderline ovarian tumors (BOTs), which are a special type of epithelial tumor of the ovary, lie between benign and malignant tumors and have low malignant potential. Due to the fact that the early symptoms of these tumors are relatively subtle, they are not easy to diagnose clinically. This study explores advancements in clinical detection methods and provides a comprehensive overview of molecules such as cell migration factors, cell transcription factors, cell damage repair factors, cell cycle regulators, and tumor suppressor genes that are related to the development of BOTs and their related mechanisms in recent years, thus aiming to provide more sensitive, specific, and efficient differential diagnosis and treatment plans for patients to improve their prognosis and survival outcomes.

Comparative analysis of microplastics detection methods applied to marine sediments: A case study in the Bay of Marseille.

An intercomparison exercise on "microplastics in sediment" was carried out by five laboratories using samples collected in the Bay of Marseille in September 2021. The results from different extraction and identification methods varied depending on the type and size classes of MPs, and was better than 80 % for the size class >300 µm and for the fragments. The variability in recovery rates can be attributed to the choice of reagents and extraction protocols. Recovery rates per laboratory were between 47 % and 113 % and the use of ZnCl2 and NaI increased recovery rates by an average of 70 %. The lowest recovery rates (47 and 53 %) were attributed to the reference methods (FTIR and LDIR), conversely the highest (80 and 87 %) were attributed to identification by Nile Red. The average ranged between 23 and 53 items /50 g d.w. with decreases offshore and at greater depth.

Development of human papillomavirus and its detection methods (Review).

Human papillomavirus (HPV) infection plays an important role in cervical cancer. HPV is classified within the Papillomaviridae family and is a non-enveloped, small DNA virus. HPV infection can be classified into two distinct scenarios: i) With or without integration into the host chromosomes. Detection of its infection can be useful in the study of cervical lesions. In the present review, the structural and functional features of HPV, HPV typing, infection and transmission mode, the risk factors for cervical susceptibility to infection and HPV detection methods are described in detail. The development of HPV detection methods may have far-reaching significance in the prevention and treatment of cervical disease. This review summarizes the advantages and limitations of each HPV detection method.

Recent research progress in tetrodotoxin detection and quantitative analysis methods.

Tetrodotoxin (TTX) is a highly potent and widely distributed ion-channel marine neurotoxin; it has no specific antidote and poses a great risk to human health. Therefore, detecting and quantifying TTX to effectively implement prevention strategies is important for food safety. The development of novel and highly sensitive, highly specific, rapid, and simple techniques for trace TTX detection has attracted widespread attention. This review summarizes the latest advances in the detection and quantitative analysis of TTX, covering detection methods based on biological and cellular sensors, immunoassays and immunosensors, aptamers, and liquid chromatography-mass spectrometry. It further discusses the advantages and applications of various detection technologies developed for TTX and focuses on the frontier areas and development directions of TTX detection, providing relevant information for further investigations.

Advances on multiclass pesticide residue determination in citrus fruits and citrus-derived products - A critical review.

The application of agrochemicals in citrus fruits is widely used to improve the quality of crops, increase production yields, and prolong post-harvest life. However, these substances are potentially toxic for humans and the ecosystem due to their widespread use, high stability, and bioaccumulation. Conventional techniques for determining pesticide residues in citrus fruits are chromatographic methods coupled with different detectors. However, in recent years, the need for analytical strategies that are less polluting for the environment has encouraged the appearance of new alternatives, such as sensors and biosensors, which allow selective and sensitive detection of pesticide residues in real time. A comprehensive overview of the analytical platforms used to determine pesticide residues in citrus fruits and citrus-derived products is presented herein. The review focuses on the evolution of these methods since 2015, their limitations, and possible future perspectives for improving pesticide residue determination and reducing environmental contamination.

A narrative review of wastewater surveillance: pathogens of concern, applications, detection methods, and challenges.

Introduction: The emergence and resurgence of pathogens have led to significant global health challenges. Wastewater surveillance has historically been used to track water-borne or fecal-orally transmitted pathogens, providing a sensitive means of monitoring pathogens within a community. This technique offers a comprehensive, real-time, and cost-effective approach to disease surveillance, especially for diseases that are difficult to monitor through individual clinical screenings. Methods: This narrative review examines the current state of knowledge on wastewater surveillance, emphasizing important findings and techniques used to detect potential pathogens from wastewater. It includes a review of literature on the detection methods, the pathogens of concern, and the challenges faced in the surveillance process. Results: Wastewater surveillance has proven to be a powerful tool for early warning and timely intervention of infectious diseases. It can detect pathogens shed by asymptomatic and pre-symptomatic individuals, providing an accurate population-level view of disease transmission. The review highlights the applications of wastewater surveillance in tracking key pathogens of concern, such as gastrointestinal pathogens, respiratory pathogens, and viruses like SARS-CoV-2. Discussion: The review discusses the benefits of wastewater surveillance in public health, particularly its role in enhancing existing systems for infectious disease surveillance. It also addresses the challenges faced, such as the need for improved detection methods and the management of antimicrobial resistance. The potential for wastewater surveillance to inform public health mitigation strategies and outbreak response protocols is emphasized. Conclusion: Wastewater surveillance is a valuable tool in the fight against infectious diseases. It offers a unique perspective on the spread and evolution of pathogens, aiding in the prevention and control of disease epidemics. This review underscores the importance of continued research and development in this field to overcome current challenges and maximize the potential of wastewater surveillance in public health.

Research progress of loop-mediated isothermal amplification in the detection of Salmonella for food safety applications.

Salmonella, the prevailing zoonotic pathogen within the Enterobacteriaceae family, holds the foremost position in global bacterial poisoning incidents, thereby signifying its paramount importance in public health. Consequently, the imperative for expeditious and uncomplicated detection techniques for Salmonella in food is underscored. After more than two decades of development, loop-mediated isothermal amplification (LAMP) has emerged as a potent adjunct to the polymerase chain reaction, demonstrating significant advantages in the realm of isothermal amplification. Its growing prominence is evident in the increasing number of reports on its application in the rapid detection of Salmonella. This paper provides a systematic exposition of the technical principles and characteristics of LAMP, along with an overview of the research progress made in the rapid detection of Salmonella using LAMP and its derivatives. Additionally, the target genes reported in various levels, including Salmonella genus, species, serogroup, and serotype, are summarized, aiming to offer a valuable reference for the advancement of LAMP application in Salmonella detection. Finally, we look forward to the development direction of LAMP and expect more competitive methods to provide strong support for food safety applications.

An overview of analytical methods for detecting endogenous hydrogen sulfide (H2S) in plants.

The significance of hydrogen sulfide (H2S) as a crucial gasotransmitter has been shown extensively in plants, and endogenous H2S is often modulated to activate H2S signaling when plants respond to numerous developmental and environmental cues. Consequently, elucidating the H2S physiological concentrations and the H2S generation intensity of plants is key to understanding the activation mechanism of H2S signaling, which has attracted increasing attention. Currently, a variety of reaction-based methods have been reported for monitoring H2S concentration in vivo and in vitro. In this review, we summarize and describe in detail several methods for quantifying and bioimaging endogenous H2S in plants systems, mainly the spectrophotometer-dependent methylene blue (MB) method and fluorescence probes, including the reaction mechanisms, design strategies, response principles, and application details. Moreover, we also summarize the advantages and disadvantages of these methods as well as the research scenarios in which they are applicable. We expect that this review will provide some guidelines on the selection of methods for H2S sensing and the comprehensive investigations into H2S signaling in plants.

Recognition of necroptosis: From molecular mechanisms to detection methods.

Necroptosis is a crucial modality of programmed cell death characterized by distinct morphological and biochemical hallmarks, including cell membrane rupture, organelle swelling, cytoplasmic and nuclear disintegration, cellular contents leakage, and release of damage-associated molecular patterns (DAMPs), accompanied by the inflammatory responses. Studies have shown that necroptosis is involved in the etiology and evolution of a variety of pathologies including organ damage, inflammation disorders, and cancer. Despite its significance, the field of necroptosis research grapples with the challenge of non-standardized detection methodologies. In this review, we introduce the fundamental concepts and molecular mechanisms of necroptosis and critically appraise the principles, merits, and inherent limitations of current detection technologies. This endeavor seeks to establish a methodological framework for necroptosis detection, thereby propelling deeper insights into the research of cell necroptosis.

Diagnosis of Alzheimer's disease: Towards accuracy and accessibility.

Alzheimer's disease (AD) is a serious dementia afflicting aging population and is characterized by cognitive decline, amyloid-ß plaques, and neurofibrillary tangles. AD substantially impairs the life quality of the victims and poses a heavy burden on the society at large. The number of people with dementia due to AD, prodromal AD, and preclinical AD is estimated to stand at roughly 3.2, 69, and 315 million worldwide, respectively. Current clinical diagnosis is based on clinical symptoms, and clinical research demonstrated that positron emission tomography (PET) and cerebrospinal fluid (CSF) biomarkers had excellent diagnostic performance. However, the application of CSF biomarker tests and PET are restricted by the invasiveness and high cost. The presence of clinical symptoms means that AD pathology has been progressing for many years, and only a few drugs have been approved for the traetemnt of AD. Therefore, early diagnosis is extremely important for controlling the outcomes caused by AD. In this review, we provided an overview of developing clinical diagnostic criteria, diagnostic strategies under clinical research, developing blood based-biomarker assays, and promising nanotechnologically-based assays.

Recent Advances on Jamming and Spoofing Detection in GNSS.

Increased interest in the development and integration of navigation and positioning services into a wide range of receivers makes them susceptible to a variety of security attacks such as Global Navigation Satellite Systems (GNSS) jamming and spoofing attacks. The availability of low-cost devices including software-defined radios (SDRs) provides a wide accessibility of affordable platforms that can be used to perform these attacks. Early detection of jamming and spoofing interferences is essential for mitigation and avoidance of service degradation. For these reasons, the development of efficient detection methods has become an important research topic and a number of effective methods has been reported in the literature. This survey offers the reader a comprehensive and systematic review of methods for detection of GNSS jamming and spoofing interferences. The categorization and classification of selected methods according to specific parameters and features is provided with a focus on recent advances in the field. Although many different detection methods have been reported, significant research efforts toward developing new and more efficient methods remain ongoing. These efforts are driven by the rapid development and increased number of attacks that pose high-security risks. The presented review of GNSS jamming and spoofing detection methods may be used for the selection of the most appropriate solution for specific purposes and constraints and also to provide a reference for future research.