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A ratiometric probe is described for the fluorometric determination of Cu(II) ions based on their quenching effect on the luminescence of dually-emitting quantum dots (QDs). ZnS QDs were doped with Mn(II) and subsequently modified with mercaptopropionic acid to give the QD probe which consists of a sole fluorophore but has two emission peaks (at 430 and 590 nm under 310 nm excitation, respectively). On addition of Cu(II) ions, the 590 nm band is quenched while the 430 nm band exhibits a little change. The changes in the intensity ratios of the yellow and the purple bands increases linearly in the 0 to 3.0 µM Cu(II) concentration range, and the detection limit reached 14 nM. The QD probe was validated and successfully applied to the determination of Cu(II) in spiked real water samples. Graphical abstract Mn-doped ZnS (ZnS:Mn(II)) quantum dots were synthesized with yellow fluorescence. After the modification of 3-mercaptopropionic acid (MPA), ZnS:Mn(II) was transferred to aqueous phase and became MPA modified Mn-doped ZnS (MPA- ZnS:Mn(II)). The fluorescence was changed to purple upon the addition of copper ions because the yellow band was largely quenched while the purple band only changed a little.
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Apoptosis is a tightly regulated cellular process that plays an essential role in the development, aging, cancer biology, immune response, and pathogenesis of various diseases. Herein, we report a new SERS sensing strategy for in vitro sensitive detection of early apoptotic cells. The principle of this method is to in situ synthesize silver nanoparticles (AgNPs) on the phosphatidylserine (PS) of the apoptotic cell membrane during the early apoptosis, which enables distinguishing normal and apoptotic cells. The total assay time of the presented method is only 10 min, thus being faster, cheaper and simpler than current techniques for the detection of apoptosis. The intrinsic mechanism was verified by different approaches based on externalized phosphatidylserine. In addition, the detection process is real-time and label-free; i.e., the intrinsic SERS spectra from the cellular membrane are directly employed for apoptosis real-time detection, which avoids using additional chemical or biological reagents as external signal indicators. Therefore, our SERS approach may serve as a potentially practical tool for sensitive and real-time detection of early cell apoptosis, complementing the state-of-the-art strategies, e.g. flow cytometry. While further investigation is required to better understand the intrinsic mechanism of the in situ coating method, the current results may provide another choice for real-time detection of early apoptosis.
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Techniques to distinguish between live and dead bacteria in a quantitative manner are in high demand in numerous fields including medical care, food safety, and public security as well as basic science research. This work demonstrates new nanostructures (silver nanoparticles coating bacteria structure, Bacteria@AgNPs) and their utility for rapid counting of live and dead bacteria by surface-enhanced Raman scattering (SERS). We found that suspensions containing Gram-negative organisms as well as AgNPs give strong SERS signals of live bacteria when generated selectively on the particle surface. However, almost no SERS signals can be detected from Bacteria@AgNPs suspensions containing dead bacteria. We demonstrate successful quantification of different percentages of dead bacteria both in bulk liquid and on glass surfaces by using SERS mapping on a single cell basis. Furthermore, different chemicals have been used to elucidate the mechanism involved in this observation. Finally, we used the Bacteria@AgNPs method to detect antibiotic resistance of E. coli strains against several antibiotics used in human medicine.
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Bactérias/isolamento & purificação , Análise Espectral Raman/métodos , Nanopartículas Metálicas , Microscopia Eletrônica de Transmissão , Prata/químicaRESUMO
This paper describes a novel double-stranded DNA detection method through resonance between SYBR Green I and DNA with the surface-enhanced resonance Raman scattering (SERRS) assay, which opens an avenue to the quantitative and reliable application of SERRS in DNA detection.
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DNA/análise , Corantes Fluorescentes/química , Compostos Orgânicos/química , Reação em Cadeia da Polimerase/métodos , Análise Espectral Raman/métodos , Benzotiazóis , Diaminas , QuinolinasRESUMO
The bio-sensing for the convenient detection of bacteria has been widely explored with the use of various sensing materials and techniques. It is still a challenge to achieve an ultrasensitive and selective, but simple, rapid, and inexpensive detection method for bacteria. We report on surface-enhanced Raman scattering (SERS) for the detection of living bacteria in drinking water by employing a synthesis of silver nanoparticles coating the cell wall of bacteria. We found that the Raman signals intensity of bacteria after AgNP synthesis mainly depends on the zeta potential of the cell wall. The enhancement of the Raman signal of bacteria using this strategy is about 30-fold higher than that in the case of a simply mixed colloid-bacterial suspension. The total assay time required is only 10 min and the total reactants' volume needed to analyze bacteria in a real environment is as low as 1 mL. Particularly, only one droplet of 3 µL sample is necessary for each SERS measurement. Furthermore, we can use this novel strategy to discriminate three strains of Escherichia coli and one strain of Staphylococcus epidermidis by hierarchy cluster analysis. Finally, we can detect bacteria down to 2.5 × 10(2) cells/mL on a hydrophobic glass slide by SERS mapping. Thus, our detection method offers prominent advantages, such as reduced assay time, simple handling, low reactant volumes, small amount of sample, and higher sensitivity and selectivity compared to previously reported label free methods. This novel strategy may be extended to open an avenue for developing various SERS-based biosensors.
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Escherichia coli/isolamento & purificação , Nanopartículas Metálicas/química , Prata/química , Análise Espectral Raman/métodos , Staphylococcus epidermidis/isolamento & purificação , Microbiologia da Água , Parede Celular/química , Análise por Conglomerados , Escherichia coli/citologia , Escherichia coli/fisiologia , Hidroxilamina/química , Viabilidade Microbiana , Nitrato de Prata/química , Staphylococcus epidermidis/citologia , Staphylococcus epidermidis/fisiologia , Propriedades de SuperfícieRESUMO
Ethyl carbamate, a by-product of fermentation and storage with widespread occurrence in fermented food and alcoholic beverages, is a compound potentially toxic to humans. In this work, a new approach for quantitative detection of ethyl carbamate in alcoholic beverages, based on surface-enhanced Raman scattering (SERS), is reported. Individual silver-coated gold nanoparticle colloids are used as SERS amplifiers, yielding high Raman enhancement of ethyl carbamate in three kinds of alcoholic beverages (vodka, Obstler, and white rum). The characteristic band at 1,003 cm(-1), which is the strongest and best reproducible peak in the SERS spectra, was used for quantitative evaluation of ethyl carbamate. The limit of detection, which corresponds to a signal-to-noise ratio of 3, was 9.0 × 10(-9) M (0.8 µg · L(-1)), 1.3 × 10(-7) M (11.6 µg · L(-1)), and 7.8 × 10(-8) M (6.9 µg · L(-1)), respectively. Surface-enhanced Raman spectroscopy offers great practical potential for the in situ assessment and identification of ethyl carbamate in the alcoholic beverage industry.
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Bebidas Alcoólicas/análise , Contaminação de Alimentos/análise , Análise Espectral Raman/métodos , Uretana/análiseRESUMO
Interferon gamma (IFN-γ), can serve as an active diagnostic biomarker of a broad spectrum of diseases such as auto inflammatory disease, viral and bacterial, parasites infections, and tumor control. The low physiological concentration of IFN-γ at pgâ§mL-1 level for most diseases such as tuberculosis and lung cancer demand highly sensitive and selective detection methods. To achieve the goal, a novel paper-based SERS aptasensor towards rapid, dual-modal (visual and ultrasensitive) detection of IFN-γ is presented for the first time. A lateral flow platform with low-cost and user-friendly format in this study is adopted. The detection relies on the competition of the specific aptamer sequence of IFN-γ between its complementary DNA in the test line and IFN-γ in the sample solution. The presence of IFN-γ can be easily observed in the test line by naked eye and detected at pgâ§mL-1 level by a portable Raman spectrometer. Linear detection range of 10-2000 pgâ§mL-1 could be obtained with detection limit of 8.7 pgâ§mL-1. In addition, as low as 10 pg/mL of IFN-γ in human serum could be detected, which is comparable with the results from ELISA.Clinical Relevance- This study establishes a simple, rapid, and low-cost assay for dual-modal detection of IFN-γ, which is in urgent demand in clinics especially vitally important in resource-limited areas.
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Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Humanos , Técnicas Biossensoriais/métodos , Interferon gamaRESUMO
Botrytis cinerea is a phytopathogenic fungus that causes gray mold, a major postharvest disease of fruits and vegetables. Chemical fungicides remain the main solution to control Botrytis disease, but concerns have raised about their safety to environment and human health, and there is an increasing need for development of more effective and less toxic treatments. In this study the divalent cation chelating agent ethylenediaminetetraacetic acid (EDTA) exhibited marked antifungal activity against B. cinerea, including inhibition of spore germination, mycelial growth, infection cushion formation, stimulation of cell death, and impairment of fungal virulence. These adverse effects of EDTA could be reversed by the addition of calcium ion, implying that metal ion chelation is involved in the fungicidal mechanism. Bean leaf and tomato fruit protection assay indicated that EDTA treatment led to a significant reduction of infection by B. cinerea. Furthermore, the antifungal activity of EDTA was significantly enhanced when used in combination with fenhexamid. These findings suggest that EDTA could be a promising tool to control B. cinerea, and application of EDTA may reduce the use of conventional chemical fungicides.
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Antifúngicos , Fungicidas Industriais , Humanos , Antifúngicos/farmacologia , Fungicidas Industriais/farmacologia , Ácido Edético/farmacologia , Botrytis , Quelantes/farmacologia , Doenças das Plantas/prevenção & controle , Doenças das Plantas/microbiologiaRESUMO
CRISPR/Cas biotechnology provides an exceptional platform for biosensor development. To date, the reported CRISPR/Cas biosensing systems have shown extraordinary performance for nucleic acids, small molecules, small proteins and microorganism detection. The CRISPR/Cas12a biosensing system, as a typical example, has been well established and applied for both nucleic acids and non-nucleic acids target detection. However, all established CRISPR/Cas12a biosensing systems are based on DNA reporters, which potentially limits further application.In this study, we established an RNA reporter based CRISPR/Cas12a biosensing system. A basic biosensing system was evaluated, and the limit of detection was found to be 1 nM. Afterwards, we optimized this biosensing system using both temperature and chemical enhancers. The final optimal biosensing system (with DTT & 37°C) shows fluorescence signal increased by a factor of ~10 compared with the basic system. The optimal biosensing system was further applied for the detection of circulating tumor DNA (ctDNA), which shows over 4 orders of magnitude detection range from 1pM to 25 nM, with the limit of detection of 1pM. This RNA reporter based CRISPR/Cas12a biosensing system provides an effective platform for nucleic acids quantification.Clinical Relevance- This research provides a novel approach for ctDNA diagnostics, which is an attractive biomarker for noninvasive monitoring of tumor growth, response, and spread.
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DNA Tumoral Circulante , Ácidos Nucleicos , RNA , Sistemas CRISPR-Cas , FluorescênciaRESUMO
Purpose: The importance of body composition on cancer outcomes is of great clinical interest. Measures of body composition that differentiate fat mass from skeletal muscle mass can help redefine our understanding of body composition for cancer survival. We investigated whether the risk of all-cause and cancer-specific mortality differ by levels of total fat mass and sarcopenia status in cancer survivors. Our secondary aim was a subgroup analysis assessing the role of race within these associations. Methods: Participants included 1682 adult cancer survivors who had undergone a dual-energy X-ray absorptiometry (DXA) examination to measure body composition, from the 1999-2006 and 2011-2018 National Health and Nutrition Examination Survey (NHANES). Total fat mass was categorized into tertiles (we assessed high vs. low tertiles), and sarcopenia was considered as having an appendicular skeletal muscle mass index less than 7.26 kg/m2 for males and less than 5.45 kg/m2 for females. Multivariable Cox proportional hazard models estimated the adjusted hazard ratio (aHR) and 95% confidence interval (CI). Results: The mean age of study participants was 61.9 years, and they were followed up for an average of 9.67 years. The prevalence of sarcopenia was 25.0% (N = 304), and 33.4% (N = 561) had a high total fat mass. Participants with a higher fat mass (aHR = 1.30, 95% CI = 1.06-1.61) and with sarcopenia (aHR = 1.51, 95% CI = 1.22-1.88) had a 30% and 51% increased risk of all-cause mortality compared to participants with a low fat mass and with no sarcopenia, respectively. Further, sarcopenia (aHR = 1.74, 95% CI = 1.23-2.29) was associated with a higher risk of cancer-specific mortality in cancer survivors. The association between sarcopenia and all-cause mortality was twice as strong in Black people (aHR = 2.99, 95% CI = 1.39-6.06) compared to White people (aHR = 1.53, 95% CI = 1.19-1.95). Conclusions: Our findings show the opposing relations of fat mass and appendicular skeletal muscle mass index with mortality in a national sample of cancer survivors, and that the relationships may differ by race. These results emphasize the importance of maintaining a healthy body composition among cancer survivors.
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BACKGROUND: Head and neck cancer (HNC) mortality differs by race, ethnicity, and socioeconomic status (SES). However, it is unclear whether the relationship between race/ethnicity and HNC-specific mortality varies according to the residence-level SES. METHODS: Data from the Surveillance Epidemiology and End Results database included participants with primary HNC between 2006 and 2017 (followed through 2018) to assess the joint association of race/ethnicity and census-tract level SES Yost-index groups (quintiles) with all-cause and HNC-specific mortalities. Relative survival rates at 1, 5, and 10 years were calculated. Multivariable Cox proportional hazard regression models estimated hazard-ratios and 95% confidence intervals for all-cause mortality, and Fine-Gray subdistribution hazard models for HNC-specific mortality. Cumulative incidence curves for HNC-specific deaths were estimated. RESULTS: 76,095 patients were included in the analysis: 63.2% were <65 years, 73.4% male, and 11.3% non-Hispanic (NH) Black. Most patients (58.3%) were diagnosed at regional or distant stages and 20.6% died of HNC. The five-year relative survival rate increased with SES group, with 51.6% in the lowest SES group, and 74.1% in the highest SES group. NH-Black patients had higher risk of all-cause and HNC-specific mortality than NH-White patients, regardless of the SES group. NH-Asian/Pacific Islander and Hispanic patients had higher risk of HNC-specific mortality in some SES groups. CONCLUSIONS: NH-Black patients of all SES strata had significantly worse outcomes. Other factors, such as healthcare quality, may be associated with persistent disparities. IMPACT: The study highlights the persistence of significant racial disparities in HNC survival across socioeconomic categories. There is need to consider additional factors underlying these disparities.
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Etnicidade , Neoplasias de Cabeça e Pescoço , Disparidades nos Níveis de Saúde , Fatores Socioeconômicos , Feminino , Humanos , Masculino , Neoplasias de Cabeça e Pescoço/etnologia , Enquadramento Interseccional , Programa de SEER , Classe Social , Grupos Raciais , Negro ou Afro-AmericanoRESUMO
Background: Allostatic load has been linked to an increased risk of death in various populations. However, to date, there is no research specifically investigating the effect of allostatic load on mortality in older cancer survivors. Aims: To investigate the association between allostatic load (AL) and mortality in older cancer survivors. Method: A total of 1,291 adults aged 60 years or older who survived for ≥1 year since cancer diagnoses were identified from the 1999-2010 National Health and Nutrition Examination Survey. AL was the exposure of interest incorporating 9 clinical measures/biomarkers; one point was added to AL if any of the measures/biomarkers exceeded the normal level. The sum of points was categorized as an ordinal variable to reflect low, moderate, and high AL. Our outcomes of interest were all-cause, cancer-specific, and cardiovascular disease (CVD)-specific mortality. Death was identified by linkage to the National Death Index. Multivariable Cox proportional hazards models were used to estimate adjusted hazard ratio (aHR) and 95% confidence intervals (CI) of mortality by AL category. Results: Overall, 53.6% of participants were male and 78.4% were white. The mean age of study participants at interview was 72.8 years (SD=7.1). A total of 546 participants died during the follow-up (median follow-up time: 8.0 years). Among them, 158 died of cancer and 106 died of cardiovascular events. Results from multivariable Cox proportional hazards models showed that higher ALS was positively associated with higher all-cause mortality (ALS=4-9 vs. ALS =0-1: aHR=1.52, 95% CI =1.17-1.98, p-trend<0.01) and higher cancer-specific mortality (ALS=4-9 vs. ALS =0-1: aHR=1.80, 95% CI =1.12-2.90, p-trend=0.01). The association between ALS and cardiovascular mortality was positive but non-significant (ALS=4-9 vs. ALS =0-1: aHR=1.59, 95% CI =0.86-2.94, p-trend=0.11). Conclusions: Our study suggests that older cancer survivors can have a higher risk of death if they have a high burden of AL.
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We sought to compare overall survival (OS) by comorbidity burden among patients with stage I/II non-small cell lung cancer (NSCLC) who received thoracoscopic resection. Utilizing data from the National Cancer Database, we conducted a survival analysis among patients aged 50+ with stage I/II NSCLC who received thoracoscopic resection between 2010 and 2017. The comorbidity burden was measured by the Charlson comorbidity index (CCI, 0, 1, 2+). Multivariable Cox proportional hazard models were used to compare overall survival relative to the CCI (CCI of 0 as the referent). Subgroup analyses were conducted considering sex, age groups, days from diagnosis to surgery, facility type, laterality, and type of surgery. For this study, 61,760 patients were included, with a mean age of 69.1 years (SD: 8.5). Notably, 51.2% had a CCI of 0, 31.8% had a CCI of 1, and 17.0% had a CCI of 2+. Most participants were non-Hispanic White (87.5%), and 56.9% were female. We found that an increase in the CCI was associated with a higher risk of all-cause mortality (CCI 1 vs. 0 aHR: 1.24, 95% CI: 1.20-1.28; CCI 2+ vs. 0 aHR: 1.51, 95% CI: 1.45-1.57; p-trend < 0.01). Our subgroup analysis according to sex suggested that the association between CCI and risk of death was stronger in women.
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OBJECTIVES: Individuals with prior cancer diagnosis are more likely to have low muscle mass (LMM) than their cancer-free counterparts. Understanding the effects of LMM on the prognosis of cancer survivors can be clinically important. The aim of this study was to investigate whether risks for all-cause and cardiovascular disease (CVD)-specific mortality differ by status of LMM in cancer survivors and a matched cohort without cancer history. METHODS: We used cohort data from the 1999-2006 and 2011-2014 National Health and Nutrition Examination Survey. Participants included 946 adults surviving for ≥1 since cancer diagnosis and a matched cohort (by age, sex, and race) without cancer history (N = 1857). LMM was defined by appendicular lean mass and body height (men <7.26 kg/m2, women <5.45 kg/m2). Death was ascertained via the National Death Index and cause of death was assessed via International Classification of Diseases, Tenth Revision. Multivariable Cox proportional hazards models were used to estimate adjusted hazard ratio (aHR) and 95% confidence interval (CI) of LMM. RESULTS: The mean age of cancer survivors and matched cohort was 60.6 y (SD 15) and 60.2 y (SD 14.9), respectively. The median follow-up was 10.5 y for survivors and 10.9 y for matched cohort. Overall, 22.2% of cancer survivors and 19.7% of the matched cohort had LMM, respectively. In all, 321 survivors (33.9%) and 495 participants (26.7%) in the matched cohort died during follow-up. CVD-specific deaths were identified in 58 survivors (6.1%) and 122 participants in the matched cohort (6.6%). The multivariable Cox model suggested that LMM was positively associated with all-cause (aHR, 1.73; 95% CI, 1.31-2.29) and CVD-specific (aHR, 2.13; 95% CI, 1.14-4.00) mortality in cancer survivors. The associations between LMM and risk for all-cause (aHR, 1.24; 95% CI, 0.98-1.56) and CVD-specific (aHR, 1.21; 95% CI, 0.75-1.93) mortality were not statistically significant in the matched cohort. CONCLUSION: Cancer survivors with LMM have an increased risk for all-cause and CVD-specific mortality. This increase appears to be larger than that in counterparts without cancer history.
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Sobreviventes de Câncer , Doenças Cardiovasculares , Neoplasias , Masculino , Adulto , Humanos , Feminino , Doenças Cardiovasculares/diagnóstico , Inquéritos Nutricionais , Prognóstico , Neoplasias/complicações , Músculos , Fatores de RiscoRESUMO
Objective: To slow down the spread of SARS-CoV-2, many countries have instituted preventive approaches (masks, social distancing) as well as the distribution of vaccines. Adherence to these preventive measures is crucial to the success of controlling the pandemic but decreased perceptions of disease severity could limit adherence. The aim of our study was to observe changes in perceived personal severity and perceived community severity; the study also explored their predictors. Methods: In a longitudinal study from an address-based probability survey in Detroit, we asked participants to rate their perceived severity of COVID-19 for themselves and for their community. In our analysis, 746 participants were queried across 5 waves of the Detroit Metro Area Communities Study surveys from March 31 to October 27 in 2020. We tested for trends in changes of self-reported perceived severity for themselves and for their community; we assessed the effects of different predictors of the two severities through mixed effects logistic regression models. Results: Our results highlight that the overall levels of perceived community and personal severity were decreasing over time even though both severities were fluctuating with rising confirmed case counts. Compared with non-Hispanic (NH) White Detroiters, NH Black Detroiters reported a higher perceived personal severity (OR: 5.30, 95% CI: 2.97, 9.47) but both groups reported similar levels of perceived community severity. We found steeper declines in perceived severity in NH White than NH Black Detroiters over time; the impact of education and income on perceived severity was attenuated in NH Black Detroiters compared with NH White Detroiters. Conclusions: Our findings suggested that perceived severity for COVID-19 decreased through time and was affected by different factors among varied racial/ethnic groups. Future interventions to slow the pace of the pandemic should take into account perceived personal and community severities among varied ethnic/racial subgroups.
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COVID-19 , Humanos , Estudos Longitudinais , Michigan/epidemiologia , Pandemias/prevenção & controle , SARS-CoV-2RESUMO
Signal amplification is crucial in developing a reliable disposable screen-printed carbon electrodes (SPCEs)-based biosensor for analyte detection with a narrow detection window. This work demonstrated a novel label-free electrochemical aptasensor based on SPCEs for the ultrasensitive detection of ochratoxin A (OTA). The graphene oxide-DNA (GO-DNA) complex as a signal amplifier with easy preparation was investigated for the first time. The proposed aptasensor based on the SPCEs/GO/cDNA-aptamer/3D-rGO-AuNPs structure was formed through the hybridization of aptamer-linked 3D-rGO/AuNPs and its complementary DNA-linked GO (GO-cDNA). The presence of OTA was discerned by its specific aptamer forming a curled OTA-aptamer complex and releasing the GO-cDNA from the surface of SPCEs. The resulting OTA-aptamer complex hindered interfacial electron transfer on the sensing surface, leading to the decreased peak current. The GO-cDNA further amplified the peak current change. This electrochemical aptasensor showed a low limit of detection of 5 fg/mL as well as good reproducibility with the relative standard deviation (RSD) of 4.38%. Moreover, the detection result of OTA in the rice and oat samples was comparable with that of the enzyme-linked immunosorbent assay (ELISA) kit. In general, the OTA aptasensor used in this work with convenient preparation, low-cost, good selectivity, high sensitivity and acceptable reproducibility can be proposed as a reliable point-of-care (POC) technique for OTA determination.
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Pseudomonas aeruginosa (P. aeruginosa), a ubiquitous opportunistic pathogen, can frequently cause chronic obstructive pulmonary disease, cystic fibrosis and chronic wounds, and potentially lead to severe morbidity and mortality. Timely and adequate treatment of nosocomial infection in clinic depends on rapid detection and accurate identification of P. aeruginosa and its early-stage antibiotic susceptibility test. Traditional methods like plating culture, polymerase chain reaction, and enzyme-linked immune sorbent assays are time-consuming and require expensive equipment, limiting the rapid diagnostic application. Advanced sensing strategy capable of fast, sensitive and simple detection with low cost has therefore become highly desired in point of care testing (POCT) of nosocomial pathogens. Within this review, advanced detection and sensing strategies for P. aeruginosa cells along with associated quorum sensing (QS) molecules over the last ten years are discussed and summarized. Firstly, the principles of four commonly used sensing strategies including localized surface plasmon resonance (LSPR), surface-enhanced Raman spectroscopy (SERS), electrochemistry, and fluorescence are briefly overviewed. Then, the advancement of the above sensing techniques for P. aeruginosa cells and its QS biomarkers detection are introduced, respectively. In addition, the integration with novel compatible platforms towards clinical application is highlighted in each section. Finally, the current achievements are summarized along with proposed challenges and prospects.
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Infecções por Pseudomonas , Pseudomonas aeruginosa , Antibacterianos , Proteínas de Bactérias , Biomarcadores , Humanos , Infecções por Pseudomonas/diagnóstico , Percepção de QuorumRESUMO
Campylobacter jejuni is the major cause of campylobacteriosis, one of the most common foodborne illnesses worldwide. Here, we report the development of RAA-exo-probe and RAA-CRIPSR/Cas12a assays for the detection of C. jejuni in food samples. The two assays were found to be highly specific to C. jejuni and highly sensitive, as they were one log more sensitive compared to the traditional culture method, with detection thresholds of 9 and 5 copies per reaction, respectively. These assays successfully detected C. jejuni in spiked chicken samples and natural meat samples (chicken, beef, mutton, etc.) and were overall less dependent on expensive equipment, only requiring a fluorescent reader. Their ease of use compared to other nucleic acid amplification-based methods indicates that these assays could be adapted for the rapid, routine surveillance of C. jejuni contamination in food samples, particularly for work done in the field or poorly equipped labs.
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Campylobacter jejuni , Análise de Alimentos , Animais , Sistemas CRISPR-Cas , Campylobacter jejuni/genética , Bovinos , Galinhas , Técnicas de Amplificação de Ácido Nucleico/métodos , Recombinases/genéticaRESUMO
Ochratoxin A (OTA) primarily obtained by the genera aspergillus and penicillium, is one of the toxic substances for different organs and systems of the human body such as the kidney, liver, neurons and the immune system. Moreover, it is considered to cause tumors and fetal malformation even at a very low concentration. Fast and sensitive assay for detection of OTA at ultralow levels in foods and agricultural products has been an increasing demand. In this study, a new label-free electrochemical biosensor based on three-dimensional reduced graphene oxide/gold nanoparticles/aptamer for OTA detection was constructed. The 3D-rGO/Au NPs nanocomposites were firstly synthesized using a one-pot hydrothermal process under optimized experimental conditions. The 3D-rGO/Au NPs with considerable particular surface area and outstanding electrical conductivity was then coated on a glass carbon electrode to provide tremendous binding sites for -SH modified aptamer via the distinctive Au-S linkage. The presence of OTA was specifically captured by aptamer and resulted in electrochemical impedance spectroscopy (EIS) signal response accordingly. The constructed impedimetric aptasensor obtained a broad linear response from 1 pg/mL to 10 ng/mL with an LOD of 0.34 pg/mL toward OTA detection, highlighting the excellent sensitivity. Satisfactory reproducibility was also achieved with the relative standard deviation (RSD) of 1.393%. Moreover, the proposed aptasensor obtained a good recovery of OTA detection in red wine samples within the range of 93.14 to 112.75% along with a low LOD of 0.023 ng/mL, indicating its applicability for OTA detection in real samples along with economical, specific, susceptible, fast, easy, and transportable merits.
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Técnicas Biossensoriais , Ocratoxinas/análise , Aptâmeros de Nucleotídeos , Carbono , Espectroscopia Dielétrica , Técnicas Eletroquímicas , Eletrodos , Ouro/química , Grafite/química , Humanos , Limite de Detecção , Nanopartículas Metálicas/química , Nanocompostos , Reprodutibilidade dos Testes , VinhoRESUMO
The rapid, highly sensitive, and accurate detection of bacteria is the focus of various fields, especially food safety and public health. Surface-enhanced Raman spectroscopy (SERS), with the advantages of being fast, sensitive, and nondestructive, can be used to directly obtain molecular fingerprint information, as well as for the on-line qualitative analysis of multicomponent samples. It has therefore become an effective technique for bacterial detection. Within this progress report, advances in the detection of bacteria using SERS and other compatible techniques are discussed in order to summarize its development in recent years. First, the enhancement principle and mechanism of SERS technology are briefly overviewed. The second part is devoted to a label-free strategy for the detection of bacterial cells and bacterial metabolites. In this section, important considerations that must be made to improve bacterial SERS signals are discussed. Then, the label-based SERS strategy involves the design strategy of SERS tags, the immunomagnetic separation of SERS tags, and the capture of bacteria from solution and dye-labeled SERS primers. In the third part, several novel SERS compatible technologies and applications in clinical and food safety are introduced. In the final part, the results achieved are summarized and future perspectives are proposed.