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Surface enhanced Raman spectroscopy (SERS) is meeting the requirements in biomedical science being a highly sensitive and specific analytical tool. By employing portable Raman systems in combination with customized sample pre-treatment, point-of-care-testing (POCT) becomes feasible. Powerful SERS-active sensing surfaces with high stability and modification layers if required are available for testing and application in complex biological matrices such as body fluids, cells or tissues. This review summarizes the current state in sample collection and pretreatment in SERS detection protocols, SERS detection schemes, i.e. direct and indirect SERS as well as targeted and non-targeted SERS, and SERS-active sensing surfaces. Moreover, the recent developments and advances of SERS in biomedical application scenarios, such as infectious diseases, cancer diagnostics and therapeutic drug monitoring is given, which enables the readers to identify the sample collection and preparation protocols, SERS substrates and detection strategies that are best-suited for their specific applications in biomedicine.
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The integration of microfabrication and microfluidics techniques into cell culture technology has significantly transformed cell culture conditions, scaffold architecture, and tissue biofabrication. These tools offer precise control over cell positioning and enable high-resolution analysis and testing. Culturing cells in 3D systems, such as spheroids and organoids, enables recapitulating the interaction between cells and the extracellular matrix, thereby allowing the creation of human-based biomimetic tissue models that are well-suited for pre-clinical drug screening. Here, we demonstrate an innovative microfluidic device for the formation, culture, and testing of hepatocyte spheroids, which comprises a large array of patterned microwells for hosting hepatic spheroid culture in a reproducible and organized format in a dynamic fluidic environment. The device allows maintaining and characterizing different spheroid sizes as well as exposing to various drugs in parallel enabling high-throughput experimentation. These liver spheroids exhibit physiologically relevant hepatic functionality, as evidenced by their ability to produce albumin and urea at levels comparable to in vivo conditions and the capability to distinguish the toxic effects of selected drugs. This highlights the effectiveness of the microenvironment provided by the chip in maintaining the functionality of hepatocyte spheroids. These data support the notion that the liver-spheroid chip provides a favorable microenvironment for the maintenance of hepatocyte spheroid functionality.
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Brucellosis is a bacterial zoonotic disease that requires major attention for both health and financial facilities in many parts of the world including the Mediterranean and the Middle East. The existing gold standard diagnosis relies on the culturing technique, which is costly and time-consuming with a duration of up to 45 days. The Brucella protease biosensor represents a new detection approach that will lead to low-cost point-of-care devices for sensitive Brucella detection. In addition, the described diagnostic device is portable and simple to operate by a nurse or non-skilled clinician making it appropriate for the low-resource setting. In this study, we rely on the total extracellular protease proteolytic activity on specific peptide sequences identified using the FRET assay by high-throughput screening from the library of peptide (96 short peptides such as dipeptides and tripeptides) substrates for Brucella melitensis (B. melitensis). The B. melitensis-specific protease substrate was utilized in the development of the paper-based colorimetric assay. Two specific and highly active dipeptide substrates were identified (FITC-Ahx-K-r-K-Ahx-DABCYL and FITC-Ahx-R-r-K-Ahx-DABCYL). The peptide-magnetic bead nanoprobe sensors developed based on these substrates were able to detect the Brucella with LOD as low as 1.5 × 102 and 1.5 × 103 CFU/mL using K-r dipeptide and R-r dipeptide substrates, respectively, as the recognition element. The samples were tested using this sensor in few minutes. Cross-reactivity studies confirmed that the other proteases extracted from closely related pathogens have no significant effect on the sensor. To the best of our knowledge, this assay is the first assay that can be used with low-cost, rapid, direct, and visual detection of B. melitensis.
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Giardia intestinalis is one of the most widespread intestinal parasites and is considered a major cause of epidemic or sporadic diarrhea worldwide. In this study, we aimed to develop a rapid aptameric diagnostic technique for G. intestinalis infection. First, the SELEX (Systematic Evolution of Ligands by Exponential Enrichment) process generated DNA aptamers specific to a recombinant protein of the parasite's trophozoite. Ten selection rounds were performed; each round, the DNA library was incubated with the target protein conjugated to Sepharose beads. Then, the unbound sequences were removed by washing and the specific sequences were eluted and amplified by Polymerase Chain Reaction (PCR). Two aptamers were selected, and the dissociation constants (Kd), were determined as 2.45 and 16.95 nM, showed their high affinity for the G. intestinalis trophozoite protein. Subsequently, the aptamer sequence T1, which exhibited better affinity, was employed to develop a label-free electrochemical biosensor. A thiolated aptamer was covalently immobilized onto a gold screen-printed electrode (SPGE), and the binding of the targeted protein was monitored using square wave voltammetry (SWV). The developed aptasensor enabled accurate detection of the G. intestinalis recombinant protein within the range of 0.1 pg/mL to 100 ng/mL, with an excellent sensitivity (LOD of 0.35 pg/mL). Moreover, selectivity studies showed a negligible cross-reactivity toward other proteins such as bovine serum albumin, globulin, and G. intestinalis cyst protein.
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Aptámeros de Nucleótidos , Técnicas Biosensibles , Técnicas Electroquímicas , Giardia lamblia , Proteínas Protozoarias , Técnica SELEX de Producción de Aptámeros , Aptámeros de Nucleótidos/química , Técnicas Biosensibles/métodos , Técnica SELEX de Producción de Aptámeros/métodos , Técnicas Electroquímicas/métodos , Proteínas Protozoarias/química , ADN de Cadena Simple/química , Giardiasis/diagnóstico , Giardiasis/parasitologíaRESUMEN
Therapeutic drug monitoring (TDM) is an important tool in precision medicine as it allows estimating pharmacodynamic and pharmacokinetic effects of drugs in clinical settings. An accurate, fast and real-time determination of the drug concentrations in patients ensures fast decision-making processes at the bedside to optimize the clinical treatment. Surface-enhanced Raman spectroscopy (SERS), which is based on the application of metallic nanostructured substrates to amplify the inherent weak Raman signal, is a promising technique in medical research due to its molecular specificity and trace sensitivity accompanied with short detection times. Therefore, we developed a SERS-based detection scheme using silicon nanowires decorated with silver nanoparticles, fabricated by means of top-down etching combined with chemical deposition, to detect the antibiotic ceftriaxone (CRO) in spiked fresh plasma and microdialysis samples. We successfully detected CRO in both matrices with an LOD of 94 µM in protein-depleted fresh plasma and 1.4 µM in microdialysate.
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Nanopartículas del Metal , Nanocables , Humanos , Antibacterianos/farmacología , Plata/química , Ceftriaxona , Silicio/química , Nanopartículas del Metal/química , Nanocables/química , Espectrometría Raman/métodosRESUMEN
Matrix metalloproteinases (MMPs) are zinc-dependent proteinases that are capable of cleavage of extracellular matrix (ECM) proteins and enzymes and play an important role in lung dysfunction. Specifically, MMP-2 is produced in the lung by alveolar epithelial and endothelial cells and other immune cells, such as macrophages. MMP-2 regulatory pathway is initiated in alveolar macrophages during acute lung injury (ALI), which may increase pulmonary inflammation. Therefore, there is a critical need for fast and reliable techniques to track the acute respiratory distress syndrome (ARDS). Here, we describe near-infrared fluorescence resonance energy transfer (NI-FRET) MMP-2-based probe for the in vivo detection of ALI induced by lipopolysaccharides (LPS). LPS-induced MMP-2 was measured using near-infrared (NIR) imaging after 1, 2, 4, 5, and 24 h of LPS exposure. Our results were compared with the data obtained from ELISA and Western blotting, demonstrating that MMP-2 fluorescence probe provide a promising in vivo diagnostic tool for ALI/ARDS in infected mice.
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For the progress of point-of-care medicine, where individual health status can be easily and quickly monitored using a handheld sensor, saliva serves as one of the best-suited body fluids thanks to its availability and abundance of physiological indicators. Salivary biomarkers, combined with rapid and highly sensitive detection tools, may pave the way to new real-time health monitoring and personalized preventative therapy branches using saliva as a target matrix. Saliva is increasing in importance in liquid biopsy, a non-invasive approach that helps physicians diagnose and characterize specific diseases in patients. Here, we propose a proof-of-concept study combining the unique specificity in biomolecular recognition provided by surface-enhanced Raman spectroscopy (SERS) in combination with molecular dynamics (MD) simulations, which give leave to explore the biomolecular absorption mechanism on nanoparticle surfaces, in order to verify the traceability of two validated salivary indicators, i.e., interleukin-8 (IL-8) and lysozyme (LYZ), implicated in oropharyngeal squamous cell carcinoma (OSCC) and oral infection. This strategy simultaneously assures the detection and interpretation of protein biomarkers in saliva, ultimately opening a new route for the evolution of fast and accurate point-of-care SERS-based sensors of interest in precision medicine diagnostics.
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Carcinoma de Células Escamosas , Neoplasias de Cabeza y Cuello , Neoplasias de la Boca , Humanos , Neoplasias de la Boca/diagnóstico , Sistemas de Atención de Punto , Carcinoma de Células Escamosas/diagnóstico , Neoplasias de Cabeza y Cuello/diagnóstico , Biomarcadores/análisis , Saliva/química , Espectrometría Raman , Biomarcadores de Tumor/análisisRESUMEN
We achieved external activation of local hot-spot sites in supracolloidal assembly structures. The concept was demonstrated by boosting surface-enhanced Raman scattering (SERS) efficiency by one order of magnitude through a heating-induced process. Our approach involves assembling gold nanoparticles with distinct dimensions, i.e. 16 and 80 nm, into well-defined planet-satellite-type arrangement structures using thermoresponsive (poly(N-isopropylacrylamide)) star polymer linkers. Insights into the assembly process were obtained by calculations within the Derjaguin-Landau-Verwey-Overbeek (DLVO) theory framework. We observe one order of magnitude increase in SERS enhancement by a heating-induced volume-phase transition. This magnification aligns with simulations run using the finite-difference time-domain (FDTD) method. The implications of this adaptive supracolloidal concept are twofold: Firstly, our approach bypasses limitations of existing systems that are associated with the limited accessibility of electromagnetic hot-spot sites in strongly coupled, static assemblies of plasmonic nanoparticles, by providing the capability of dynamic hot-spot re-configuration. Second, these externally activated probes offer promising opportunities for the development of messenger materials and associated sensing strategies.
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Sepsis is an immune response to a microbial invasion that causes organ injury and dysfunction due to a systemic inflammatory response. Sepsis is a serious, life-threatening condition and a widely recognized global health challenge. Given its high death rate, it is critical to diagnose sepsis and start treatment as early as possible. There is an urgent need for a sensitive and rapid screening method for detecting sepsis. In this study, we investigated the use of MMP-9 as a biomarker for sepsis. A colorimetric paper-based biosensor was used for the detection of MMP-9 utilizing peptide-magnetic nanoparticle conjugates. The method is based on the cleavage of the MMP-9-specific peptide by the protease leading to the detaching of the magnetic beads from the sensor surface and changing of color. A fecal intraperitoneal (FIP) challenge was used to induce sepsis in mice, and an MMP-9 secretion was measured by taking blood and Bronchoalveolar Lavage (BAL) fluid samples at 1 h, 2 h, 4 h, and 20 h (early sepsis) post-challenge intervals. The results of the paper-based sensor for the detection of MMP-9 levels in blood samples and BAL samples were compared with ELISA and Western Blot. We found that both blood and BAL levels of MMP-9 increased immediately and could be detected as early as 1 h in FIP mice post-challenge. Our work adds evidence to the assertion that MMP-9 is a reliable biomarker for the detection of sepsis at early stages.
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Metaloproteinasa 9 de la Matriz , Sepsis , Animales , Ratones , Sepsis/diagnóstico , Biomarcadores , Colorimetría , Modelos Animales de EnfermedadRESUMEN
Acute respiratory distress syndrome (ARDS) is a worldwide health concern. The pathophysiological features of ALI/ARDS include a pulmonary immunological response. The development of a rapid and low-cost biosensing platform for the detection of ARDS is urgently needed. In this study, we report the development of a paper-based multiplexed sensing platform to detect human NE, PR3 and MMP-2 proteases. Through monitoring the three proteases in infected mice after the intra-nasal administration of LPS, we showed that these proteases played an essential role in ALI/ARDS. The paper-based sensor utilized a colorimetric detection approach based on the cleavage of peptide-magnetic nanoparticle conjugates, which led to a change in the gold nanoparticle-modified paper sensor. The multiplexing of human NE, PR3 and MMP-2 proteases was tested and compared after 30 min, 2 h, 4 h and 24 h of LPS administration. The multiplexing platform of the three analytes led to relatively marked peptide cleavage occurring only after 30 min and 24 h. The results demonstrated that MMP-2, PR3 and human NE can provide a promising biosensing platform for ALI/ARDS in infected mice at different stages. MMP-2 was detected at all stages (30 min-24 h); however, the detection of human NE and PR3 can be useful for early- (30 min) and late-stage (24 h) detection of ALI/ARDS. Further studies are necessary to apply these potential diagnostic biosensing platforms to detect ARDS in patients.
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Nanopartículas del Metal , Síndrome de Dificultad Respiratoria , Humanos , Animales , Ratones , Líquido del Lavado Bronquioalveolar , Lipopolisacáridos , Metaloproteinasa 2 de la Matriz , Oro , Síndrome de Dificultad Respiratoria/diagnóstico , Biomarcadores , Péptido HidrolasasRESUMEN
The early detection of Parkinson's disease (PD) can significantly improve treatment and quality of life in patients. 5-S-Cysteinyl-dopamine (CDA) is a key metabolite of high relevance for the early detection of PD. Therefore, its sensitive detection with fast and robust methods can improve its use as a biomarker. In this work we show the potentialities of label-free SERS spectroscopy in detecting CDA in aqueous solutions and artificial biofluids, with a simple, fast and sensitive approach. We present a detailed experimental SERS band assignment of CDA employing silver nanoparticle (AgNP) substrates in aqueous media, which was supported by theoretical calculations and simulated Raman and SERS spectra. The tentative orientation of CDA over the AgNP was also studied, indicating that catechol and carboxylic acid play a key role in the metallic surface adsorption. Moreover, we showed that SERS can allow us to identify CDA in aqueous media at low concentration, leading to the identification of some of its characteristic bands in pure water and in synthetic cerebrospinal fluid (SCSF) below 1 × 10-8 M, while its band identification in simulated urine (SUR) can be reached at 1 × 10-7 M. In conclusion, we show that CDA can be suitably detected by means of label-free SERS spectroscopy, which can significantly improve its sensitive detection for further analytical studies as a novel biomarker and further clinical diagnosis in PD patients.
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Nanopartículas del Metal , Enfermedad de Parkinson , Humanos , Dopamina , Espectrometría Raman/métodos , Nanopartículas del Metal/química , Calidad de Vida , Plata/química , Agua , BiomarcadoresRESUMEN
This paper describes the use of cyclodextrins (CDs) to improve the determination of fluoroquinolone antibiotics in human body fluids using surface-enhanced Raman spectroscopy (SERS). CDs were used to (i) prepare the CD-SERS substrate (synthesis and stabilization of silver nanoparticles), (ii) increase the sensitivity of the assay by enhancing the interaction between analyte molecules and the substrate, and (iii) improve the analysis accuracy by reducing the interaction between the substrate and endogenous components of body fluids. Two native CDs (α-CD and ß-CD) and two of their derivatives with hydroxypropyl groups were tested, and the best results were obtained with CD-SERS substrate prepared using native ß-CD. The CD-SERS assay has been developed and optimized for the determination of commonly used and structurally related fluoroquinolones (ciprofloxacin, norfloxacin, pefloxacin, and levofloxacin) in urine and blood plasma samples. Importantly, the non-significant difference in the interaction of the CD-modified SERS substrate with various fluoroquinolones has been successfully used to develop a versatile assay suitable for the analyte-class-specific analysis. Calibration plots were obtained for concentration ranges suitable for the determination of the antibiotics in urine (50-500 µg mL-1) and blood plasma (1-6 µg mL-1). The following figures of merit were obtained (for urine and blood plasma, respectively): RSD values are ≤15% and ≤23%, LOD values are 2.9-5.8 and 0.05-0.34 µg mL-1, recovery ranges are 96-105% and 91-111%. In addition, the influence of excessive concentrations of some main endogenous components of the body fluids on the analytical signal was studied. This step was used to evaluate possible limitations of the assay associated with the deviation of the composition of the body fluid matrix. Therefore, accounting for the short analysis time (≤15 min) and the use of a portable Raman spectrometer, the proposed assay can be suggested for therapeutic drug monitoring in hospitals.
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Líquidos Corporales , Ciclodextrinas , Nanopartículas del Metal , Humanos , Nanopartículas del Metal/química , Plata/química , Antibacterianos/orina , Espectrometría Raman/métodos , Fluoroquinolonas , PlasmaRESUMEN
A Raman-based label-free analytical method was developed to detect the antibiotic ciprofloxacin (CIP) in various pharmaceutical formulations in the presence of different matrices (e.g., ear drops, eye drops and infusion solutions). The Raman spectral analysis is performed for semiquantification in the case of a low background Raman signal, i.e., the signal originating from the excipient and carrier substance of the formulation does not interfere with the fingerprint spectrum of ciprofloxacin. In the case of a background spectrum rich in Raman modes originating from the excipient and carrier substance of the formulation, the pharmaceutical formulation is diluted 1:5000, and thus, the background signal is undetectable. Due to the high affinity of ciprofloxacin towards metallic surfaces, surface-enhanced Raman spectroscopy (SERS) is applied to allow for the sensitive detection of ciprofloxacin. Moreover, the developed measurement routine can be applied to monitor the degradation of the active component ciprofloxacin within the pharmaceutical formulation. The developed assay can therefore be extended to the pharmaceutical industry for quality control in assay applications or the preparation of individualized medicines.
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Ciprofloxacina , Excipientes , Antibacterianos/análisis , Ciprofloxacina/análisis , Composición de Medicamentos , Soluciones Oftálmicas , Espectrometría Raman/métodosRESUMEN
Aptamers are RNA/DNA oligonucleotide molecules that specifically bind to a targeted complementary molecule. As potential recognition elements with promising diagnostic and therapeutic applications, aptamers, such as monoclonal antibodies, could provide many treatment and diagnostic options for blood diseases. Aptamers present several superior features over antibodies, including a simple in vitro selection and production, ease of modification and conjugation, high stability, and low immunogenicity. Emerging as promising alternatives to antibodies, aptamers could overcome the present limitations of monoclonal antibody therapy to provide novel diagnostic, therapeutic, and preventive treatments for blood diseases. Researchers in several biomedical areas, such as biomarker detection, diagnosis, imaging, and targeted therapy, have widely investigated aptamers, and several aptamers have been developed over the past two decades. One of these is the pegaptanib sodium injection, an aptamer-based therapeutic that functions as an anti-angiogenic medicine, and it is the first aptamer approved by the U.S. Food and Drug Administration (FDA) for therapeutic use. Several other aptamers are now in clinical trials. In this review, we highlight the current state of aptamers in the clinical trial program and introduce some promising aptamers currently in pre-clinical development for blood diseases.
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Aptámeros de NucleótidosRESUMEN
Numerous approaches have been proposed to overcome the intrinsically low selectivity of surface-enhanced Raman spectroscopy (SERS), and the modification of SERS substrates with diverse recognition molecules is one of such approaches. In contrast to the use of antibodies, aptamers, and molecularly imprinted polymers, application of cyclodextrins (CDs) is still developing with less than 100 papers since 1993. Therefore, the main goal of this review is the critical analysis of all available papers on the use of CDs in SERS analysis, including physicochemical studies of CD complexation and the effect of CD presence on the Raman enhancement. The results of the review reveal that there is controversial information about CD efficiency and further experimental investigations have to be done in order to estimate the real potential of CDs in SERS-based analysis.
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Dopamine (DA) regulates several functions in the central nervous system and its depletion is responsible for psychological disorders like Parkinson's disease. Several analytical approaches have been presented for DA detection in pathological diagnosis. SERS spectroscopy is a highly promising technique for the sensitive detection of DA. However, an improvement in its detection in aqueous solution is highly desirable for reliable quantification in biological fluids. In this work, we explored a label-free SERS approach for DA detection, employing two conventional methods to synthesize Ag colloids: reduction via citrates (c-AgNPs) and reduction via hydroxylamine (h-AgNPs), and SERS measurements were performed with a laser at 488 nm wavelength. Under these conditions, DA was identified through reproducible SERS spectra in the c-AgNP medium; however, the SERS spectra of DA in h-AgNP solution showed a completely different SERS profile. SERS band analysis revealed that DA in h-AgNPs was oxidized and converted into polydopamine (PDA), which was triggered after exposure to laser radiation. DA oxidation and PDA formation were followed over time through the SERS band profile at pH 7, 9 and 12. We found that in situ PDA formation started after 50 min of laser irradiation of DA at pH 7, while DA was quickly oxidized at pH 9 and 12. Here, we present a detailed SERS band analysis of PDA, which sheds light on the molecular steps in the pathway formation of the PDA structure. Spectroscopic analysis and characterization revealed that a long laser exposure time led to the formation of stable PDA complexes with AgNPs, which allowed us to propose a novel approach for synthesis of AgNP-PDA composites. In conclusion, to detect DA through a label-free SERS approach, c-AgNPs must be employed, while stable AgNP-PDA materials can be achieved with h-AgNPs and 488 nm laser excitation.
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Dopamina/química , Dopamina/síntesis química , Nanopartículas del Metal/química , Plata/química , Estructura Molecular , Polimerizacion , Espectrometría Raman , Propiedades de SuperficieRESUMEN
This study demonstrates the development of a sensitive, specific, and quantitative peptide-based nanoprobe prototype assay for the detection of Legionellaceae in a simple way and in a short time. In this work, proteases present in the culture supernatants of Legionella spp. were used as a biomarker. Fluorogenic peptide substrates, specific to Legionella strains culture supernatant proteases, were identified. Peptidases produced a significant increase in the fluorescence intensity following the cleavage of the dipeptide fluorogenic substrates. The specific substrates were identified and coupled with carboxyl-terminated nano-magnetic particles (NMPs). On the other hand, the C-terminal was conjugated with the cysteine residue to covalently integrate with a gold sensing platform via the Au-S linkage. Four different sensors were fabricated from the four specific substrates, which were treated with the protesase of six different species of Legionella. In the presence of specific protease, the peptide sequence is digested and the magnetic nanobeads moved out of the gold surface, resulting in the apparence of gold color. One of the nanoprobes sensitivity detects as low as 60 CFU mL-1 of Legionella anisa, Legionella micdadei, and Fluoribacter dumoffii. The cross-reactivity of the sensors was tested using other closely associated bacterial species and no significant cross-reactivity of the sensors was found. It is envisaged that this assay could be useful for screening purposes or might be supportive for the fast and easy detection of Legionella protease activity for water monitoring purposes.
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Técnicas Biosensibles , Legionellaceae , Legionella , PéptidosRESUMEN
The timely diagnosis of MRSA in clinical samples helps to reduce the attendant morbidity/mortality associated with infection due to the organism. The early institution of appropriate therapy or deployment of infection control protocols are dependent on a timely report from the microbiology laboratory. Various assays currently used in the identification of MRSA are associated with inherent shortcomings, thus there is a need to explore newer diagnostic frontiers that can eliminate some of these short comings at a relatively cheap, timely, specific and sensitive manner. We present in this study a MRSA specific optical immunosensor to detect the presence of the pathogen on contaminated surface using control and patient strains. Results revealed a detection limits of 103 CFU mL-1 upon visual observation, and 29 CFU mL-1 as determined by the linear regression equation, following the use of ImageJ to quantify activated cotton swab color intensity. The specificity of the sensor was examined by blind testing a panel of non-MRSA bacteria (E. coli, S. aureus and S. epidermis). Negative visual read-out was observed for all tested non-specific bacteria except for MRSA. Assay takes an average of 5 min and presents a powerful point-of-care diagnostic platform for the detection of MRSA.
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Técnicas Biosensibles , Staphylococcus aureus Resistente a Meticilina , Infecciones Estafilocócicas , Colorimetría , Escherichia coli , Humanos , Inmunoensayo , Sensibilidad y Especificidad , Infecciones Estafilocócicas/diagnóstico , Staphylococcus aureusRESUMEN
There is mounting evidence that contaminated hospital environment plays a crucial role in the transmission of nosocomial pathogens such as MRSA. The institution of infection control protocols is predicated on the early laboratory detection of the pathogen from relevant samples. Processing of environmental samples for the presence of bacterial contaminants in the clinical environment is poorly standardized when compared with analysis of clinical samples. The various laboratory methods available for processing environmental samples are difficult to standardized and most require a long turnaround time before results are available. In this study, we present a report of the performance of a novel pathogen aptasensor swab designed to qualitatively and quantitatively detect MRSA, on contaminated non-absorbable surfaces. The visual detection limit of the MRSA aptasensor swab was less than 100 CFU/ml and theoretically using a standard curve, was 2 CFU/ml. A relatively short turnaround time of 5 min was established for the assay while the linear range of quantitation was 102-105 CFU/ml. Engineered aptasensor targets MRSA selectively and binds to none of the other tested bacterial pathogen, on a multi-contaminated surface. This novel detection tool was easy to use and relatively cheap to produce.