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Bacterial infection is a common impediment towards wound healing. Detecting bacterial infections is important to promote wound healing and curb chronic non-healing wounds. In this review, we firstly discuss bacterial communities, including aerobic and anaerobic bacteria in various types of wounds. Following the discussion of wound sampling methods (swab, biopsy) for different wounds, we then discuss laboratory based conventional methods (bacteria cultures, Gram staining, analytical profile index systems, polymerase chain reaction, and gas chromatography coupled with mass spectrometry), focusing on their recent improvement. After that we discussed the contemporary biosensor methods, including e-Nose, electrochemical sensors, surface enhanced Raman spectroscopy, and nucleic acid lateral flow immunoassay. Biosensors embedded into wound dressing, termed wearable sensors or smart wound dressing, are also discussed for their ability of enabling bacteria detection directly from wound sites without the need for obtaining swab/biopsy samples. We have compared all the detection methods for their performance according to their respective targets (either bacteria cells or volatile/non-volatile metabolites); after that we evaluate the suitability of various methods in providing timely and accurate diagnostic results towards real-time, point-of-care testing of bacterial infections.
Assuntos
Infecções Bacterianas , Técnicas Biossensoriais , Dispositivos Eletrônicos Vestíveis , Infecção dos Ferimentos , Bactérias , Infecções Bacterianas/diagnóstico , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Infecção dos Ferimentos/diagnóstico , Infecção dos Ferimentos/microbiologiaRESUMO
A portable surface-enhanced Raman spectroscopy (SERS) sensor for detecting pyocyanin (PYO) in simulated wound fluid and from bacteria samples was developed. Solution-phase SERS detection protocols are designed to be compatible with two different clinical practices for wound exudate collection, namely negative pressure liquid collection and swabbing. For citrate-coated metal nanoparticles of three different compositions, i.e. gold (AuNPs), alloyed silver/gold (AgAuNPs), and silver (AgNPs), we firstly confirmed their interaction with PYO in the complex wound fluid, using fluorescence quenching experiments, which rationalized the Raman enhancement effects. We then demonstrated the Raman enhancement effects of the metal nanoparticles in the order of AgNPs > AgAuNPs > AuNPs. The limit of detection (LOD) achieved for PYO is 1.1 µM (in a linear range of 0.1-25 µM by the AgNPs), 10.9 µM (in a linear range of 5-100 µM, by the AgAuNPs), and 17.7 µM (in a linear range of 10-100 µM by the AuNPs). The AgNP and AgAuNP sensors together cover the sensitivity and dynamic range requirements for the clinical detection of wound infection, where PYO is present at a concentration of 1-50 µM. In addition, sterilized cotton swabs were used to collect wound fluid and transfer samples into AgNP solution for SERS measurements. This detection protocol was completed within 5 minutes with a LOD of 23.1 µM (in a linear range of 15-100 µM). The SERS sensing protocol was validated by its successful detection of PYO in cultured Pseudomonas aeruginosa bacteria. The findings presented in this work pave the way towards point-of-care diagnostics of wound infections.
Assuntos
Nanopartículas Metálicas , Piocianina , Ouro , Prata , Análise Espectral RamanRESUMO
Acetylation of histones, the major protein component of eukaryotic chromosomes, contributes to the epigenetic regulation of gene expression and is also involved in cancer development. A recent study revealed the correlation between tumour formation and acetylation level of lysine K18 on histone H3. In this study, we developed two colorimetric in vitro assays using gold nanoparticles (AuNPs) for identification of lysine K18 acetylation on histone H3 peptide. In assay I, citrate ion-capped AuNP without further modification was employed. Simply mixing the K18 peptide with AuNP solution leads to distinct particle aggregation, relative to that by non-acetylated or lysine K14 acetylated control peptides. In assay II, an AuNP-peptide-antibody composite was synthesized and used as both the sensing probe and the transducing element. By mixing the sample peptides with the composite solution followed by PBS screening, different aggregation behaviours were observed between the K18 acetylated target peptide and the control sequences. Both assays are capable of identifying the acetylated peptides, and also differentiating the distinctive acetylation positions that differ merely by a distance of three amino acids.
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Técnicas Biossensoriais , Histonas/química , Lisina/química , Nanopartículas Metálicas/química , Peptídeos/química , Processamento de Proteína Pós-Traducional , Acetilação , Sequência de Aminoácidos , Anticorpos/química , Cromatina/química , Cromatina/metabolismo , Colorimetria/métodos , Colorimetria/normas , Floculação , Ouro/química , Histonas/metabolismo , Lisina/metabolismo , Peptídeos/síntese química , Peptídeos/metabolismoRESUMO
Forkhead box protein 1 (FoxA1) is a member of the forkhead family of winged helix transcription factors that plays pivotal roles in the development and differentiation of multiple organs and in the regulation of estrogen-stimulated genes. Conventional analytical methods-electrophoretic mobility shift assay (EMSA) and fluorescence anisotropy (FA)-as well as a gold nanoparticles (AuNPs)-based assay were used to study DNA binding properties of FoxA1 and ligand interruption of FoxA1-DNA binding. In the AuNPs assay, the distinct ability of protein-DNA complex to protect AuNPs against salt-induced aggregation was exploited to screen sequence selectivity and determine the binding affinity constant based on AuNPs color change and absorbance spectrum shift. Both conventional EMSA and FA and the AuNPs assay suggested that FoxA1 binds to DNA in a core sequence-dependent manner and the flanking sequence also played a role to influence the affinity. The EMSA and AuNPs were found to be more sensitive than FA in differentiation of sequence-dependent affinity. With the addition of a spin filtration step, AuNPs assay has been extended for studying small molecular ligand inhibition of FoxA1-DNA interactions enabling drug screening. The results correlate very well with those obtained using FA.
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Sondas de DNA/metabolismo , Ensaio de Desvio de Mobilidade Eletroforética , Polarização de Fluorescência , Ouro/química , Fator 3-alfa Nuclear de Hepatócito/metabolismo , Nanopartículas Metálicas/química , Sequência de Bases , Sondas de DNA/química , Fator 3-alfa Nuclear de Hepatócito/antagonistas & inibidores , Fator 3-alfa Nuclear de Hepatócito/genética , Humanos , Ligantes , Proteínas Ligantes de Maltose/genética , Proteínas Ligantes de Maltose/metabolismo , Ligação Proteica , Proteínas Recombinantes de Fusão/biossíntese , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Bibliotecas de Moléculas Pequenas/química , Bibliotecas de Moléculas Pequenas/metabolismoRESUMO
Total bacterial count in food is one of important food safety criteria. The current plate count method (Heterotrophic Plate Count) for food analysis requires microbiology lab facilities and at least 2â days turnover time. We developed a rapid fluorescence-based total bacterial count method that utilises semiconductor nanorods (SNRs) conjugated with a lectin Griffonia simplicifolia II (GSII-SNRs) to stain bacterial cells captured on syringe filters, via the common N-acetylglucosamine molecules on bacterial cell wall. This "Filter-and-Stain" detection method has a rapid turnover time of 20â min. The fluorescence emission can be seen under UV light with minimum interference from food sample background. The fluorescence intensity quantified through image analysis is proportional to the bacterial concentration with a limit of detection of 1000â CFU/mL, for total bacterial count assessment in food safety. Moreover, the GSII-SNRs do not bind to heat inactivated bacterial cells, and thus can differentiate live and dead bacteria. Our method has been validated with representative food (coffee powder, raw spinach leaves, and ready-to-eat tomato salsa) to demonstrate its high potential for on-site food safety assessment, especially in places with no immediate access to microbiology labs.
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Nanotubos , Semicondutores , Nanotubos/química , Microbiologia de Alimentos , Carga Bacteriana , Lectinas/química , Análise de Alimentos , Spinacia oleracea/química , Spinacia oleracea/microbiologia , Solanum lycopersicum/química , Solanum lycopersicum/microbiologiaRESUMO
Cortisol is a key stress biomarker in humans and animals, including fishes. In aquafarming, stress monitoring using cortisol quantification can help to optimize aquaculture practices for welfare and productivity enhancement. However, most current methods for cortisol detection rely on invasive tissue sampling. In this work, we developed a gold nanoparticle (AuNP)-based cortisol sensor to address the demand of detecting picomolar ranges of cortisol from complex fish tank water matrices as a non-invasive alternative for more effective stress monitoring. We first identified a DNA aptamer with effective binding to cortisol and then conjugated the thiol-labelled aptamer to AuNPs together with a blocker molecule (CALNN) to form an Au-Apt-CALNN conjugate that is stable in fish tank water. The cortisol detection principle is based on magnesium chloride (MgCl2)-induced particle aggregation, where the cortisol-bound aptamer on the AuNPs folds into a tertiary structure and provides greater protection for Au-Apt-CALNN against MgCl2-induced aggregation due to steric stabilization. At an optimum MgCl2 concentration, the differential stability of particles with and without cortisol binding offers a limit of detection (LOD) of 100 pM for cortisol within a 35 min reaction. The aptasensor has been validated on recirculating aquaculture system (RAS) fish tank water samples by the HPLC method and was able to detect changes in water cortisol induced by two different stress paradigms. This on-site deployable and non-invasive sensor offers opportunities for more efficient and real-time fish stress monitoring for the optimization of aquaculture practices.
Assuntos
Aptâmeros de Nucleotídeos , Técnicas Biossensoriais , Ouro , Hidrocortisona , Nanopartículas Metálicas , Ouro/química , Nanopartículas Metálicas/química , Hidrocortisona/análise , Hidrocortisona/metabolismo , Animais , Aptâmeros de Nucleotídeos/química , Técnicas Biossensoriais/métodos , Peixes , Estresse Fisiológico , Limite de DetecçãoRESUMO
In situ analysis of sweat biomarkers potentially provides noninvasive lifestyle monitoring and early diagnosis. Quantitative detection of sweat rate is crucial for thermoregulation and preventing heat injuries. Here, a skin-attachable paper fluidic patch is reported for in situ colorimetric sensing of multiple sweat markers (pH, glucose, lactate, and uric acid) with concurrent sweat rate tracking. Two sets of fluidic patterns-multiplexed detection zones and a longitudinal sweat rate channel-are directly printed by an automated ink dispenser from a specially developed ceramic-based ink. The ceramic ink thermal-cures into an impervious barrier, confining sweat within the channels. The ceramic-ink-printed boundary achieves higher pattern resolution, prevents fluid leakage, attains pattern thermal stability, and resistant to organic solvents. The cellulose matrix of the detection zones is modified with nanoparticles to improve the color homogeneity and sweat sensor sensitivity. The sweat rate channel is made moisture sensitive by incorporating a metal-salt-based dye. The change in saturation/color of the detection zones and/or channels upon sweat addition can be visually detected or quantified by a smartphone camera. A cost-effective way is provided to fabricate paper fluidic sensor patches, successfully demonstrating on-body multiplexed evaluation of sweat analytes. Such skin wearables offer on-site analysis, meaningful to an increasingly health-conscious population.
Assuntos
Técnicas Biossensoriais , Dispositivos Eletrônicos Vestíveis , Suor , Colorimetria , Tinta , GlucoseRESUMO
Total bacterial count is a routine parameter in microbial safety assessment used in many fields, such as drinking water and industrial water testing. The current gold standard method for counting bacteria is the plate culture method (or heterotrophic plate count) that requires a microbiology laboratory and a long turnover time of at least 24 hours. To tackle these shortcomings, we developed a rapid total bacterial count method that relies on gold nanoparticles (AuNPs) conjugated with affinity ligands to stain bacterial cells captured on a syringe filter. Two affinity ligands were exploited, i.e. a DNA aptamer (AB2) and a lectin Griffonia simplicifolia II (GSII) that recognize bacterial cell wall commonalities, i.e. peptidoglycan and its amino sugars. Upon proper formulation with addition of a surfactant, the AB2 conjugated AuNPs (AB2-AuNPs) can selectively stain bacterial cells captured on the filter membrane with a higher sensitivity than GSII-AuNPs. Measuring the staining intensity using an in-house-built handheld detector allowed us to correlate its intensity reading with the total number of bacterial units present. This bacteria quantification method, referred to as "Filter-and-Stain", had an efficient turnover time of 20 min suggesting its potential usage for rapid on-site applications. Additionally, the detection sensitivity provided by the AB2-AuNP nanoreagent offered a limit of detection as low as 100 CFU mL-1. We have demonstrated the use of the AB2-AuNPs for detection of bacteria from environmental water samples.
Assuntos
Aptâmeros de Nucleotídeos , Nanopartículas Metálicas , Qualidade da Água , Carga Bacteriana , Ouro , Bactérias , Limite de DetecçãoRESUMO
Wound healing is a dynamic process with multiple phases. Rapid profiling and quantitative characterization of inflammation and infection remain challenging. We report a paper-like battery-free in situ AI-enabled multiplexed (PETAL) sensor for holistic wound assessment by leveraging deep learning algorithms. This sensor consists of a wax-printed paper panel with five colorimetric sensors for temperature, pH, trimethylamine, uric acid, and moisture. Sensor images captured by a mobile phone were analyzed by neural network-based machine learning algorithms to determine healing status. For ex situ detection via exudates collected from rat perturbed wounds and burn wounds, the PETAL sensor can classify healing versus nonhealing status with an accuracy as high as 97%. With the sensor patches attached on rat burn wound models, in situ monitoring of wound progression or severity is demonstrated. This PETAL sensor allows early warning of adverse events, which could trigger immediate clinical intervention to facilitate wound care management.
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Queimaduras , Cicatrização , Ratos , Animais , Aprendizado de Máquina , AlgoritmosRESUMO
In this paper, we report a study of using cholesteric liquid crystals (CLCs) doped with dodecylamine for detecting aldehyde vapors. CLCs doped with dodecylamine show color change from red to yellow-green upon exposure to 300 ppmv pentyl aldehyde within 60 s. In contrast, no colorimetric response is observed when pure CLCs are used. Characterization by using FT-IR shows that the possible mechanism responsible for the colorimetric response is the formation of an imine bond between dodecylamine and pentyl aldehyde. A new C=N peak at 1670 cm(-1) appears in the spectrum after the exposure to aldehyde vapors. The CLCs doped with dodecylamine show good selectivity for pentyl aldehyde; they do not respond to 200 ppmv pentyl alcohol, pentylamine, acetone, ethanol, and water vapor. This study demonstrates the potential applications of doped CLCs as low-cost and portable gas sensors.
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Monoclonal antibodies (mAbs) for treatment of human diseases are typically human or humanized Immunoglobulin G (IgG) produced in mammalian cell lines. A rapid, less tedious, and high throughput method to quantify mAbs is in demand to accelerate mAb production efficiency. To quantify mAb titer, we developed gold nanoparticle (AuNPs)-based "mix and measure" fluorimetric assays by exploiting AuNPs' fluorescence quenching ability. The AuNPs are functionalized by an Fc binding protein, i. e. protein G, which binds human IgG and fluorescently labeled rat IgG (Alexa Fluor 488-rat IgG) with differential affinity. The assays can be in competition or displacement format. The competitive binding of human IgG drug and the labelled rat IgG to protein G-coated AuNP lead to varied fluorescent intensity that is proportional to the amount of human IgG analte; or the displacement of the labelled rat IgG from protein G-coated AuNP by human IgG can lead to fluorescent recovery that is also proportionally related to human IgG concentration. The assays can quantify therapeutic mAbs in the range of 10-1,000â mg/L, demonstrated for Herceptin, Avastin, and Humira in cell culture media. The assays have fast turn over time (within 15â min). They can be performed in microplates and are suitable for high throughput "on-line" or "at-line" measurement in mAbs production lines.
Assuntos
Anticorpos Monoclonais/análise , Fluorometria , Ouro/química , Nanopartículas Metálicas/química , HumanosRESUMO
Rapid and inexpensive immunodiagnostic assays to monitor severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) seroconversion are essential for conducting large-scale COVID-19 epidemiological surveillance and profiling humoral responses against SARS-CoV-2 infections or immunizations. Herein, a colorimetic serological assay to detect SARS-CoV-2 IgGs in patients' plasma was developed using short antigenic epitopes conjugated to gold nanoparticles (AuNPs). Four immunodominant linear B-cell epitopes, located on the spike (S) and nucleocapsid (N) proteins of SARS-CoV-2, were characterized for their IgG binding affinity and used as highly specific biological motifs on the nanoparticle to recognize target antibodies. Specific bivalent binding between SARS-CoV-2 antibodies and epitope-functionalized AuNPs trigger nanoparticle aggregation, which manifests as a distinct optical transition in the AuNPs' plasmon characteristics within 30 min of antibody introduction. Co-immobilization of two epitopes improved the assay sensitivity relative to single-epitope AuNPs with a limit of detection of 3.2 nM, commensurate with IgG levels in convalescent COVID-19-infected patients. A passivation strategy was further pursued to preserve the sensing response in human plasma medium. When tested against 35 clinical plasma samples of varying illness severity, the optimized nanosensor assay can successfully identify SARS-CoV-2 infection with 100% specificity and 83% sensitivity. As the epitopes are conserved within the circulating COVID-19 variants, the proposed platform holds great potential to serve as a cost-effective and highly specific alternative to classical immunoassays employing recombinant viral proteins. These epitope-enabled nanosensors further expand the serodiagnostic toolbox for COVID-19 epidemiological study, humoral response monitoring, or vaccine efficiency assessment.
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COVID-19 , Nanopartículas Metálicas , Humanos , SARS-CoV-2 , Ouro , COVID-19/diagnóstico , Glicoproteína da Espícula de Coronavírus/química , Epitopos , Anticorpos Antivirais , Imunoglobulina G , Sensibilidade e EspecificidadeRESUMO
In this paper, we report the utility of a colorimetric polymer stabilized cholesteric liquid crystal (PSCLC) array for detecting vaporous amines. The PSCLC with various polymer concentrations (5-20% w/w) is made into an array which shows distinct color changes upon exposure to 400 parts-permillion (ppm) octylamine vapor at 23-35 degrees C. Interestingly, PSCLC shows stronger response to primary amine over secondary amine, tertiary amine, ester, aldehyde, and alkane having similar molecular weights. PSCLCs also give detection limits as low as 2 ppmv for decylamine. Because PSCLC is transparent at room temperature and changes color upon exposure to amine vapors, it can be coated on windows or safety goggles to offer protection against amine vapors.
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Particulate matters (PMs), e. g. dusts, fibres, smokes, fumes, mists, liquid droplets and airborne respirable solid or liquid particles, are the major sources of air pollution concerning outdoor and indoor air quality. Among various PMs, bioaerosols are airborne particles that are either living organisms (bacteria, viruses, and fungi) or originate from living organisms (endotoxin, allergen, etc). PMs and/or bioaerosols have adverse health effects of infection, allergy, and irritation. Proper management and source identification of PMs and bioaerosols will reduce their negative health impact. In this review, we will discuss the analytical technologies and sensors for PMs and bioaerosols. We will first introduce four types of PM analysers, namely, filter-based gravimetric method (GMM), optical method, ß-ray absorption method (BAM), and tapered element oscillating microbalance (TEOM). We will provide examples of how commercial PM analyzers of different principles have been compared and calibrated for specific applications under different climate conditions of specific geographic locations. For bioaerosols, having more complex biological and biochemical identity, we will start from air sampling techniques, followed by a discussion of various detection methods (plate culture, molecular methods, immunoassays and biosensors) in association with compatible sampling technologies. Using Influenza A (H1â N1) virus and SARS-CoV-2 (COVID-19) virus as examples, we have highlighted air sampling and detection challenges for viral aerosols relative to bacterial and fungal aerosols. Finally, we provide a perspective for future trends according to the limitation of current commercial products and the key challenges in this field.
Assuntos
Aerossóis/análise , Poluição do Ar/prevenção & controle , Material Particulado/análise , Bactérias/isolamento & purificação , Fungos/isolamento & purificação , Vírus da Influenza A Subtipo H1N1/isolamento & purificação , SARS-CoV-2/isolamento & purificação , Esporos Fúngicos/isolamento & purificaçãoRESUMO
In aquaculture industry, fish, shellfish, and aquatic plants are cultivated in fresh, salt, or brackish waters. The increasing demand of aquatic products has stimulated the rapid growth of aquaculture industries. How to effectively monitor and control water quality is one of the key concerns for aquaculture industry to ensure high productivity and high quality. There are four major categories of water quality concerns that affect aquaculture cultivations, namely, (1) physical parameters, e.g., pH, temperature, dissolved oxygen, and salinity, (2) organic contaminants, (3) biochemical hazards, e.g., cyanotoxins, and (4) biological contaminants, i.e., pathogens. While the physical parameters are affected by climate changes, the latter three are considered as environmental factors. In this review, we provide a comprehensive summary of sensors, biosensors, and analytical technologies available for monitoring aquaculture water quality. They include low-cost commercial sensors and sensor network setups for physical parameters. They also include chromatography, mass spectrometry, biochemistry, and molecular methods (e.g., immunoassays and polymerase chain reaction assays), culture-based method, and biophysical technologies (e.g., biosensors and nanosensors) for environmental contamination factors. According to the different levels of sophistication of various analytical techniques and the information they can provide (either fine fingerprint, highly accurate quantification, semiquantification, qualitative detection, or fast screening), we will comment on how they may be used as complementary tools, as well as their potential and gaps toward current demand of real-time, online, and/or onsite detection.
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Detection of microorganisms and microbial toxins is important for health and safety. Due to their unique physical and chemical properties, nanomaterials have been extensively used to develop biosensors for rapid detection of microorganisms with microbial cells and toxins as target analytes. In this paper, the design principles of nanomaterials-based biosensors for four selected analyte categories (bacteria cells, toxins, mycotoxins, and protozoa cells), closely associated with the target analytes' properties is reviewed. Five signal transducing methods that are less equipment intensive (colorimetric, fluorimetric, surface enhanced Raman scattering, electrochemical, and magnetic relaxometry methods) is described and compared for their sensory performance (in term oflimit of detection, dynamic range, and response time) for all analyte categories. In the end, the suitability of these five sensing principles for on-site or field applications is discussed. With a comprehensive coverage of nanomaterials, design principles, sensing principles, and assessment on the sensory performance and suitability for on-site application, this review offers valuable insight and perspective for designing suitable nanomaterials-based microorganism biosensors for a given application.
Assuntos
Toxinas Bacterianas/isolamento & purificação , Técnicas Biossensoriais , Nanoestruturas/química , Campylobacter jejuni/isolamento & purificação , Campylobacter jejuni/patogenicidade , Legionella pneumophila/isolamento & purificação , Legionella pneumophila/patogenicidade , Salmonella typhimurium/isolamento & purificação , Salmonella typhimurium/patogenicidade , Análise Espectral RamanRESUMO
Cellulase is an important enzyme for hydrolyzing cellulose to form glucose. To recycle cellulase after the reaction, cellulase is often immobilized on solid supports but its activity is also compromised. In this study, we show a new hybrid cellulase aggregate with a silica core, which is prepared by physical adsorption of cross-linked cellulase on a highly porous solid support silica gel. The hybrid cellulase aggregate exhibits highest activity at pH 4.8 and 51°C, similar to the optimum condition of free cellulase. This hybrid cellulase aggregate can produce 3.4 g/L of glucose within 2 h, which is two times higher than glucose produced by using cross-linked cellulase aggregate alone (without silica core). Another advantage of the hybrid cellulase aggregate is that it can settle down naturally after the hydrolysis of cellulose, thanks to the presence of the silica core. To show its practical applications, we also study the hydrolysis of palm oil fiber by using the hybrid cellulase aggregate. Up to 5.0 g/L of glucose can be produced within 24h, and this process can be repeated five times with only 19% decrease in activity.
Assuntos
Biomassa , Celulase/química , Celulose/química , HidróliseRESUMO
Monitoring spatial distribution of chemicals in microfluidic devices by using traditional sensors is a challenging task. In this paper, we report utilization of a thin layer of cholesteric liquid crystal for monitoring ethanol inside microfluidic channels. This thin layer can be either a polymer dispersed cholesteric liquid crystal (PDCLC) layer or a free cholesteric liquid crystal (CLC) layer separated from the microfluidic device by using a thin film of PDMS. They both show visible colorimetric responses to 4% of ethanol solution inside the microfluidic channels. Moreover, the spatial distribution of ethanol inside the microfluidic channel can be reflected as a color map on the CLC sensing layers. By using this device, we successfully detected ethanol produced from fermentation taking place inside the microfluidic channel. These microfluidic channels with embedded PDCLC or embedded CLC offer a new sensing solution for monitoring volatile organic compounds in microfluidic devices.