Approach For Simultaneous Detection Research Articles

Multiplex detection of five common respiratory pathogens from bronchoalveolar lavages using high resolution melting curve analysis

BackgroundThe study describes the application of the multiplex high-resolution melting curve (MHRM) assay for the simultaneous detection of five common bacterial pathogens (Pseudomonas aeruginosa, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii and Escherichia coli) directly from bronchoalveolar lavage samples.ResultsOur MHRM assay successfully identified all five respiratory pathogens in less than 5 h, with five separate melting curves with specific melt peak temperatures (Tm). The different Tm were characterized by peaks of 78.1 ± 0.4 °C for S. aureus, 83.3 ± 0.1 °C for A. baumannii, 86.7 ± 0.2 °C for E. coli, 90.5 ± 0.1 °C for K. pneumoniae, 94.5 ± 0.2 °C for P. aeruginosa. The overall sensitivity and specificity of MHRM were 100% and 88.8–100%, respectively.ConclusionsOur MHRM assay offers a simple and fast alternative to culture approach for simultaneous detection of five major bacterial lower respiratory tract infection pathogens. Utilization of this assay can help clinicians initiate prompt and appropriate antimicrobial treatment, towards reducing the morbidity and mortality of severe respiratory infections.

Electrochemistry of rGO-Cu3H2Mo2O10 cuboidal nanostructures: An effective detection of neurotransmitter dopamine in blood serum sample

In this report, lindgrenite Cu3H2Mo2O10 cuboidal nanostructures (CMNs) and their nanocomposites with reduced graphene oxide have been explored for sensitive and selective detection of dopamine. These nanostructures have been characterized by PXRD, FESEM, TEM, Raman and XPS techniques to confirm the structural, morphological, and surface properties of rGO-CMNs nanocomposites. Electrochemical performance of rGO-CMNs nanocomposite has been investigated using CV, LSV, IMPE and EIS techniques. The lower detection limit of DA was attained to be 0.158 μΜ with the range of linearity of 1.0–10.0 μM and 34–125 μM using DPV technique. Further, the proof of concept of DA detection with serotonin selectivity has also been studied through simultaneous detection approach. The present sensing system possesses good sensitivity and selectivity against ascorbic acid and serotonin. Present sensing system was also successfully applied to detect dopamine in human blood serum sample.

Smart cationic coordination polymer: A single-crystal-to-single-crystal approach for simultaneous detection and removal of perchlorate in aqueous media

As a kind of novel persistent pollutant, perchlorate has been received a great deal of attention on toxicology transfer, degradation mode and remediation, making it an urgent task to exploit smart multifunctional material for simultaneous adsorption and detection of ClO4− in aqueous media. Here, a water-stable cationic porous coordination polymer (named as 1 ∩ NO3) has been rationally designed and constructed to achieve this goal. The resultant 1 ∩ NO3 exhibits fast sorption kinetics (30 min) and excellent capacity (108.70 mg/g) toward ClO4− anion with good repeatability and high selectivity. Luckily, the structure of ClO4−-loaded framework can be obtained directly via SC-SC approach, which proves the obvious spatial confinement of 1 ∩ NO3 towards ClO4− anion. Moreover, the fluorescence of 1 ∩ NO3 has dependent change with the concentration of ClO4−, making it as an excellent sensor with superior selective and sensitive (17.4 μg/L). It is much lower than the acceptable limit (24.5 μg/L) in drinking water standard of the US Environmental Protection Agency (EPA). These indicate that the smart adsorbent 1 ∩ NO3 is expected to be used in the synchronous detection and adsorption of ClO4− in aqueous solutions, providing a viable option in water quality monitoring and wastewater treatment.

A Novel Online Four-Dimensional SEC×SEC-IM×MS Methodology for Characterization of Monoclonal Antibody Size Variants.

The determination of size variants is a major critical quality attribute of a therapeutic monoclonal antibody (mAb that may affect the drug product safety, potency, and efficacy. Size variant characterization often relies on size-exclusion chromatography (SEC), which could be hampered by difficult identification of peaks. On the other hand, mass spectrometry (MS)-based techniques performed in nondenaturing conditions have proven to be valuable for mAb-related compound characterization. On the basis of the observation that limited SEC performance was observed in nondenaturing MS compatible ammonium acetate buffer compared with classical phosphate salts, a multidimensional analytical approach was proposed. It combines comprehensive online two-dimensional chromatography (SEC×SEC), with ion mobility and mass spectrometry (IM-MS) in nondenaturing conditions for the characterization of a variety of mAbs. We first exemplify the versatility of our approach for simultaneous detection, identification, and quantitation of adalimumab size variants. Benefits of the SEC×SEC-native IM×MS were further highlighted on forced degraded pembrolizumab and bevacizumab samples, for which the 4D setup was mandatory to obtain an extensive and unambiguous identification, and accurate quantitation of unexpected high/low molecular weight species (HMWS and LMWS). In this specific context, monomeric conformers were detected by IM-MS as HMWS or LMWS. Altogether, our results emphasize the power of comprehensive 2D LC×LC setups hyphenated to IM×MS in nondenaturing conditions with unprecedented performance including: (i) maintaining optimal SEC performance (under classical nonvolatile salt conditions), (ii) performing online native MS identification, and (iii) providing IM-MS conformational characterization of all separated size variants.

Development of a NanoBioAnalytical platform for "on-chip" qualification and quantification of platelet-derived microparticles

Blood microparticles (MPs) are small membrane vesicles (50–1000nm), derived from different cell types. They are known to play important roles in various biological processes and also recognized as potential biomarkers of various health disorders. Different methods are currently used for the detection and characterization of MPs, but none of these methods is capable to quantify and qualify total MPs at the same time, hence, there is a need to develop a new approach for simultaneous detection, characterization and quantification of microparticles. Here we show the potential of surface plasmon resonance (SPR) method coupled to atomic force microscopy (AFM) to quantify and qualify platelet-derived microparticles (PMPs), on the whole nano-to micro-meter scale. The different subpopulations of microparticles could be determined via their capture onto the surface using specific ligands. In order to verify the correlation between the capture level and the microparticles concentration in solution, two calibration standards were used: Virus-Like Particles (VLPs) and synthetic beads with a mean diameter of 53nm and 920nm respectively. The AFM analysis of the biochip surface allowed metrological analysis of captured PMPs and revealed that more than 95% of PMPs were smaller than 300nm. Our results suggest that our NanoBioAnalytical platform, combining SPR and AFM, is a suitable method for a sensitive, reproducible, label-free characterization and quantification of MPs over a wide concentration range (≈107 to 1012 particles/mL; with a limit of detection (LOD) in the lowest ng/µL range) which matches with their typical concentrations in blood.

Multi‐class obstacle detection and classification using stereovision and improved active contour models

Existing in-vehicle sensing systems are concentrated on obstacle detection for pedestrian or vehicle. Limited work has been conducted on multi-class obstacle classification. This study addresses on this issue and aims to develop an approach for simultaneous detection and classification of multi-class obstacles. Stereovision is first used to segment obstacles from traffic background, then an improved active contour model is adopted to extract complete contour curve of the detected obstacles. Based on the contour extracted, geometrical features including aspect ratio, area ratio and height are integrated for classifying object types including vehicle, pedestrian and other obstacles. The approach was tested on substantial urban traffic images and the corresponding results prove the effectiveness of the proposed approach.

Discussion of “Novel Sensor Clustering–Based Approach for Simultaneous Detection of Stiffness and Mass Changes Using Output-Only Data” by Qipei Mei and Mustafa Gül

Discussion of “Novel Sensor Clustering–Based Approach for Simultaneous Detection of Stiffness and Mass Changes Using Output-Only Data” by Qipei Mei and Mustafa Gül

Closure to “Novel Sensor Clustering–Based Approach for Simultaneous Detection of Stiffness and Mass Changes Using Output-Only Data” by Qipei Mei and Mustafa Gül

Closure to “Novel Sensor Clustering–Based Approach for Simultaneous Detection of Stiffness and Mass Changes Using Output-Only Data” by Qipei Mei and Mustafa Gül

N-acetyltransferase 2 genotyping: an accurate and feasible approach for simultaneous detection of the most common NAT2 alleles.

N -Acetyltransferase 2 (NAT2; EC 2.3.1.5) is involved in the detoxification of numerous xenobiotics. The human NAT2 gene is highly polymorphic and represents one of the best studied examples of the large interindividual variability of genetic control of drug or xenobiotic metabolism. In addition to the antituberculosis agent isoniazid (1), the first NAT2 substrate discovered, numerous other chemicals possessing a primary aromatic amine or hydrazine group, such as sulfamethazine, procainamide, hydralazine (2), dapsone, nitrazepam, and caffeine, are metabolized by this enzyme. Arylamine chemicals such as benzidine, β-naphthylamine, and 2-aminofluorene can also be acetylated by NAT2. As early as 1970, Reidenberg et al.(3) reported clinical problems associated with polymorphic acetylation of procainamide. The determination of NAT2 genotype or NAT2 phenotype has been proposed as a way to predict adverse reactions in patients with tuberculosis and before the concomitant administration of procainamide and phenytoin (4). The proportions of rapid and slow phenotypes vary in different ethnic groups. In Caucasians, 40–70% of individuals have the slow acetylator phenotype, whereas Asian populations have only 10–30% slow acetylators (5). Several single-nucleotide polymorphisms have been identified in the human NAT2 coding region, which are responsible for the observed phenotypes [see Ref. (6); accession no. X14672]. The presence of the NAT2 * 4 (wild-type) allele defines the NAT2 genotype as rapid, and combinations of the frequently occurring mutant alleles NAT2 * 5B , * 5C , * 6A , * 7B , and * 14 cluster as slow. Allele frequencies in Caucasians are 23% for NAT2 * 4 (wild type), 1.3% for * 5A , 35% for * 5B , 5.0% for * 5C , 30.5% for * 6A , 4.5% for * 7B , and 0.2% for …