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Clinical Infectious Diseases : An... Apr 2020Treatment of subarachnoid neurocysticercosis (NCC) is complicated, and assays that can guide treatment are not widely available. The reproducibility and scalability of...
A Novel, Highly Sensitive Quantitative Polymerase Chain Reaction Assay for the Diagnosis of Subarachnoid and Ventricular Neurocysticercosis and for Assessing Responses to Treatment.
BACKGROUND
Treatment of subarachnoid neurocysticercosis (NCC) is complicated, and assays that can guide treatment are not widely available. The reproducibility and scalability of molecular-based biomarkers would be of great use.
METHODS
The Taenia solium genome was mined and primers and probes were designed to target repeats with the highest coverage; the most sensitive, specific, and efficient repeat (TsolR13) was selected for clinical testing. We tested 46 plasma samples and 36 cerebral spinal fluid (CSF) samples taken from patients with subarachnoid or ventricular disease using quantitative polymerase chain reaction (qPCR).
RESULTS
The analytic sensitivity of TsolR13 was 97.3% at 240 attograms (ag) of T. solium genomic DNA and 100% analytic specificity. The clinical sensitivity in detecting active subarachnoid or ventricular disease in symptomatic patients was 100% in CSF and 81.3% in plasma. The predictive ability to distinguish active from cured disease was better for CSF (94.4% of those cured had negative qPCR results) than for plasma (86.7% of those cured tested negative). Some subjects also had plasma DNA detectable intermittently for years after being cured. Overall, the test performance was equivalent to T. solium antigen detection.
CONCLUSIONS
A qPCR test for the detection of the highly repetitive Tsol13 sequence has been developed and shown to be highly sensitive and specific for NCC, but also useful as a test of cure in CSF and for the definitive diagnosis of NCC in plasma.
Topics: Animals; Antigens, Helminth; Enzyme-Linked Immunosorbent Assay; Humans; Neurocysticercosis; Polymerase Chain Reaction; Reproducibility of Results; Sensitivity and Specificity; Taenia solium
PubMed: 31232448
DOI: 10.1093/cid/ciz541 -
Analytical Chemistry Oct 2023Owing to the exceedingly small mass involved, complete elemental characterization of single nanoparticles demands a highly precise control of signal background and noise...
Owing to the exceedingly small mass involved, complete elemental characterization of single nanoparticles demands a highly precise control of signal background and noise sources. LIBS has demonstrated remarkable merits for this task, providing a unique tool for the multielemental analysis of particles on the attogram-picogram mass scale. Despite this outstanding sensitivity, the air plasma acting as a heat source for particle dissociation and excitation is a meddling agent, often limiting the acquisition of an accurate sample signature. Although thermal effects associated with ultrashort laser pulses are known to be reduced when compared to the widely used nanosecond pulse duration regime, attempts to improve nanoinspection performance using ultrafast excitation have remained largely unexplored. Herein, picosecond laser pulses are used as a plasma excitation source for the elemental characterization of single nanoparticles isolated within optical traps in air at atmospheric pressure. Results for picosecond excitation of copper particles lead to a mass detection limit of 27 attogram, equivalent to single particles 18 nm in diameter. Temporally and wavelength-resolved plasma imaging reveals unique traits in the mechanism of atomic excitation in the picosecond regime, leading to a deeper understanding of the interactions occurring in single nanoparticle spectroscopy.
PubMed: 37729543
DOI: 10.1021/acs.analchem.3c01376 -
Journal of Microbiological Methods Jul 2020Paper-based nucleic acid testing techniques are increasingly in demand. Hence, we have developed a simple and cheap paper fluidic device to detect multiple gene targets...
Paper-based nucleic acid testing techniques are increasingly in demand. Hence, we have developed a simple and cheap paper fluidic device to detect multiple gene targets in Leptospira. Fluidic channels of the penta-clover device are drawn using a correction pen on Whatman filter paper 1. The fluid blocks the pores of the paper, avoiding leakage and ensuring the equal flow of sample to the reaction pads. The target genes are amplified by performing Loop-Mediated Isothermal Amplification (LAMP) with dry reaction components. Thecolor change of leuco crystal violetallows real-time monitoring of a positive amplification. The difference in color intensity is captured with a smartphone and analyzed using image processing software. The device amplifies the target within 15 min, detects the pathogen at a concentration as low as 50 attogram μL, detects Leptospira in blood samples without prior treatment and differentiates the Leptospira species even after 21 days of storage at room temperature.
Topics: Humans; Leptospira; Leptospirosis; Molecular Diagnostic Techniques; Nucleic Acid Amplification Techniques; Paper; Sensitivity and Specificity
PubMed: 32473300
DOI: 10.1016/j.mimet.2020.105962 -
Electroanalysis Sep 2020A sensitive detection of extremely toxic phenylpyrazole insecticide, 'Fipronil' is presented. Currently, the advancement of approaches for the detection of insecticides...
A sensitive detection of extremely toxic phenylpyrazole insecticide, 'Fipronil' is presented. Currently, the advancement of approaches for the detection of insecticides at low concentrations with less time is important for environmental safety assurance. Considering this fact, an effort has been made to develop an electrospun CoZnO nanofiber (NF) based label-free electrochemical system for the detection of fipronil. The CoZnO NF were characterized using different techniques including field emission scanning electron microscopy (FE-SEM), Energy Dispersive X-Ray Analysis (EDX), X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), and Raman Spectroscopy. Based on the experimental results, the proposed platform displayed a linear response for fipronil in the attogram/mL range despite the multiple interfering agents. The sensitivity of the device was found to be 3.99 Kῼ (g/ml) cm. Limit of detection (LOD) and limit of quantification (LOQ) were calculated and found to be 112 ag mL and 340 ag mL respectively. Further, this proposed sensor will be implemented in the fields for the rapid and proficient detection of the real samples.
PubMed: 33456276
DOI: 10.1002/elan.202000051 -
Talanta Feb 2024Inorganic gunshot residue (IGSR) has certain environmental and occupational interferent-particle sources known to display similar morphologies and elemental compositions...
Inorganic gunshot residue (IGSR) has certain environmental and occupational interferent-particle sources known to display similar morphologies and elemental compositions to IGSR. These interferences can make detecting and identifying IGSR particles difficult, especially when IGSR particle number concentrations are low. Here, single-particle inductively coupled plasma time-of-flight mass spectrometry (spICP-TOFMS) is used to explore the particle types measured from IGSR and three important interferent-particle sources: brake pads, fireworks, and mineral sunscreen. spICP-TOFMS offers results in as little as 2 min per sample. With spICP-TOFMS, the mass of most elements, down to the 10s of attograms, can be detected and quantified in individual particles with diameters from 10s to 100s of nm. At this size range, almost all interferent sources produce particles with elemental compositions that overlap with ASTM-defined particle compositions used for identifying leaded and lead-free IGSR. We establish probabilities for detecting IGSR-like particles from interference sources through the analysis of thousands of particles from each interference. Based on this analysis, robust sample-specific 'characteristic' particle types can be used to classify leaded and unleaded IGSR particles, even in the presence of interferent particles. Of the interference sources studied, particles from brake pads and fireworks are most similar to leaded IGSR; however, IGSR particles could be unequivocally classified based on detection of lead and antimony. Particles from mineral sunscreen are most similar to those from lead-free IGSR; however, lead-free IGSR particles exhibit a unique titanium-zinc-copper elemental fingerprint that is not detected in mineral sunscreen particles. Within mixtures of interference particles and IGSR, IGSR is accurately identified with limited false positives, even when the number of interference particles is over 200-times greater than that of IGSR. Our results suggest that spICP-TOFMS is a useful approach for rapid and accurate IGSR identification even in samples with high concentrations of interferent background particles.
PubMed: 37918252
DOI: 10.1016/j.talanta.2023.125368 -
Physical Review Letters Mar 2022We propose a search for low mass dark matter particles through momentum recoils caused by their scattering from trapped, nanometer-scale objects. Our projections show...
We propose a search for low mass dark matter particles through momentum recoils caused by their scattering from trapped, nanometer-scale objects. Our projections show that even with a modest array of femtogram-mass sensors, parameter space beyond the reach of existing experiments can be explored. The case of smaller, attogram-mass sensors is also analyzed-where dark matter can coherently scatter from the entire sensor-enabling a large enhancement in the scattering cross-section relative to interactions with single nuclei. Large arrays of such sensors have the potential to investigate new parameter space down to dark matter masses as low as 10 keV. If recoils from dark matter are detected by such sensors, their inherent directional sensitivity would allow an unambiguous identification of a dark matter signal.
PubMed: 35333080
DOI: 10.1103/PhysRevLett.128.101301 -
Journal of Chromatography. A Sep 2019The application of two-dimensional liquid chromatography (2D-LC) is gradually growing also in the area of metabolite profiling and identification. The current...
The application of two-dimensional liquid chromatography (2D-LC) is gradually growing also in the area of metabolite profiling and identification. The current contribution describes a heartcut 2D-UHPLC configuration that is applied in support of drug metabolism studies in development. The setup applies four LC columns: two analytical UHPLC columns to perform the first and second dimension separations, which are both preceded by a short HPLC column operated as trapping column. The first HPLC column allows a significant online preconcentration by large volume injection. The second short HPLC column is placed between the first and second dimension columns and enables the selection of orthogonal conditions in the second dimension independent of the first dimension making the heartcutting 2D approach more generic. The value of the setup was demonstrated with selective ultraviolet chromatograms obtained for the two major hydroxylated metabolites of atorvastatin separating them from a very high biological background, originating from an injection of 4 mL feces extract, by heartcut 2D-LC. In a second application, the main metabolite of imipramine was baseline separated from some minor metabolites that were co-eluting in the first dimension, allowing accurate and sensitive quantification. A quantification limit in the attogram/mL range was achieved thanks to the injection of 200 mL diluted urine, corresponding to 100 mL urine on column.
Topics: Animals; Atorvastatin; Chemistry Techniques, Analytical; Chromatography, High Pressure Liquid; Dogs; Feces; Humans; Imipramine; Pharmaceutical Preparations; Urine
PubMed: 31047660
DOI: 10.1016/j.chroma.2019.04.064 -
Biomedicines Apr 2021This work demonstrates the quantitative assay of cardiac Troponin I (cTnI), one of the key biomarkers for acute cardiovascular diseases (the leading cause of death...
This work demonstrates the quantitative assay of cardiac Troponin I (cTnI), one of the key biomarkers for acute cardiovascular diseases (the leading cause of death worldwide) using the fluorescence-based sandwich immune reaction. Surface plasmon coupled emission (SPCE) produced by non-radiative coupling of dye molecules with surface plasmons being excitable via the reverse Kretschmann format is exploited for fluorescence-based sandwich immunoassay for quantitative detection of cTnI. The SPCE fluorescence chip utilizes the gold (2 nm)-silver (50 nm) bimetallic thin film, with which molecules of the dye Alexa 488 (conjugated with detection antibodies) make a near field coupling with the plasmonic film for SPCE. The experimental results find that the SPCE greatly improves the sensitivity via enhancing the fluorescence signal (up to 50-fold) while suppressing the photo-bleaching, permitting markedly enhanced signal-to-noise ratio. The limit of detection of 21.2 ag mL (atto-gram mL) is obtained, the lowest ever reported to date amid those achieved by optical technologies such as luminescence and label-free optical sensing techniques. The features discovered such as ultrahigh sensitivity may prompt the presented technologies to be applied for early diagnosis of cTnI in blood, particularly for emergency medical centers overloaded with patients with acute myocardial infarction who would suffer from time-delayed diagnosis due to insufficient assay device sensitivity.
PubMed: 33919217
DOI: 10.3390/biomedicines9050448 -
Nanoscale Sep 2019The breathing modes of single suspended gold nanoplates have been examined by transient absorption microscopy. These vibrational modes show very high quality factors...
The breathing modes of single suspended gold nanoplates have been examined by transient absorption microscopy. These vibrational modes show very high quality factors which means that their frequencies can be accurately measured. Measurements performed before and after removing the organic layer that coats the as synthesized nanoplates show significant increases in frequency, which are consistent with removal of a few nm of organic material from the nanoplate surface. Experiments were also performed after depositing polymer beads on the sample. These measurements show a decrease in frequency in the region of the beads. This implies that adding a localized mass to the nanoplate hybridizes the vibrational normal modes, creating a new breathing mode which has a maximum amplitude at the bead. The nanoplate resonators have a mass sensing detection limit of ca. 10 attograms, which is comparable to the best results that have been achieved with plasmonic nanoparticles.
PubMed: 31453600
DOI: 10.1039/c9nr05940g