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1.
Nano Lett ; 24(4): 1214-1222, 2024 Jan 31.
Artículo en Inglés | MEDLINE | ID: mdl-38230628

RESUMEN

The ultimate sensitivity of field-effect-transistor (FET)-based devices for ionic species detection is of great interest, given that such devices are capable of monitoring single-electron-level modulations. It is shown here, from both theoretical and experimental perspectives, that for such ultimate limits to be approached the thermodynamic as well as kinetic characteristics of the (FET surface)-(linker)-(ion-receptor) ensemble must be considered. The sensitivity was probed in terms of optimal packing of the ensemble, through a minimal charge state/capacitance point of view and atomic force microscopy. Through the fine-tuning of the linker and receptor interaction with the sensing surface, a record limit of detection as well as specificity in the femtomolar range, orders of magnitude better than previously obtained and in excellent accord with prediction, was observed.

2.
BMC Microbiol ; 24(1): 31, 2024 Jan 20.
Artículo en Inglés | MEDLINE | ID: mdl-38245666

RESUMEN

BACKGROUND: Although the spread of antimicrobial resistance (AMR) through food and its production poses a significant concern, there is limited research on the prevalence of AMR bacteria in various agri-food products. Sequencing technologies are increasingly being used to track the spread of AMR genes (ARGs) in bacteria, and metagenomics has the potential to bypass some of the limitations of single isolate characterization by allowing simultaneous analysis of the agri-food product microbiome and associated resistome. However, metagenomics may still be hindered by methodological biases, presence of eukaryotic DNA, and difficulties in detecting low abundance targets within an attainable sequence coverage. The goal of this study was to assess whether limits of detection of ARGs in agri-food metagenomes were influenced by sample type and bioinformatic approaches. RESULTS: We simulated metagenomes containing different proportions of AMR pathogens and analysed them for taxonomic composition and ARGs using several common bioinformatic tools. Kraken2/Bracken estimates of species abundance were closest to expected values. However, analysis by both Kraken2/Bracken indicated presence of organisms not included in the synthetic metagenomes. Metaphlan3/Metaphlan4 analysis of community composition was more specific but with lower sensitivity than the Kraken2/Bracken analysis. Accurate detection of ARGs dropped drastically below 5X isolate genome coverage. However, it was sometimes possible to detect ARGs and closely related alleles at lower coverage levels if using a lower ARG-target coverage cutoff (< 80%). While KMA and CARD-RGI only predicted presence of expected ARG-targets or closely related gene-alleles, SRST2 (which allows read to map to multiple targets) falsely reported presence of distantly related ARGs at all isolate genome coverage levels. The presence of background microbiota in metagenomes influenced the accuracy of ARG detection by KMA, resulting in mcr-1 detection at 0.1X isolate coverage in the lettuce but not in the beef metagenome. CONCLUSIONS: This study demonstrates accurate detection of ARGs in synthetic metagenomes using various bioinformatic methods, provided that reads from the ARG-encoding organism exceed approximately 5X isolate coverage (i.e. 0.4% of a 40 million read metagenome). While lowering thresholds for target gene detection improved sensitivity, this led to the identification of alternative ARG-alleles, potentially confounding the identification of critical ARGs in the resistome. Further advancements in sequencing technologies providing increased coverage depth or extended read lengths may improve ARG detection in agri-food metagenomic samples, enabling use of this approach for tracking clinically important ARGs in agri-food samples.


Asunto(s)
Antibacterianos , Farmacorresistencia Bacteriana , Animales , Bovinos , Antibacterianos/farmacología , Farmacorresistencia Bacteriana/genética , Límite de Detección , Bacterias/genética , Genes Bacterianos/genética , Metagenoma , Metagenómica/métodos , Biología Computacional
3.
BMC Microbiol ; 24(1): 68, 2024 Feb 28.
Artículo en Inglés | MEDLINE | ID: mdl-38413863

RESUMEN

OBJECTIVES: In the current study, for the first time, we reported a novel HCV molecular diagnostic approach termed reverse transcription loop-mediated isothermal amplification integrated with a gold nanoparticles-based lateral flow biosensor (RT-LAMP-AuNPs-LFB), which we developed for rapid, sensitive, specific, simple, and visual identification of HCV. METHODS: A set of LAMP primer was designed according to 5'untranslated region (5'UTR) gene from the major HCV genotypes 1b, 2a, 3b, 6a, and 3a, which are prevalent in China. The HCV-RT-LAMP-AuNPs-LFB assay conditions, including HCV-RT-LAMP reaction temperature and time were optimized. The sensitivity, specificity, and selectivity of our assay were evaluated in the current study. The feasibility of HCV-RT-LAMP-AuNPs-LFB was confirmed through clinical serum samples from patients with suspected HCV infections. RESULTS: An unique set of HCV-RT-LAMP primers were successfully designed targeting on the 5'UTR gene. The optimal detection process, including crude nucleic acid extraction (approximately 5 min), RT-LAMP reaction (67℃, 30 min), and visual interpretation of AuNPs-LFB results (~ 2 min), could be performed within 40 min without specific instruments. The limit of detection was determined to be 20 copies per test. The HCV-RT-LAMP-AuNPs-LFB assay exhibited high specificity and anti-interference. CONCLUSIONS: These preliminary results confirmed that the HCV-RT-LAMP-AuNPs-LFB assay is a sensitive, specific, rapid, visual, and cost-saving assay for identification of HCV. This diagnostic approach has great potential value for point-of-care (POC) diagnostic of HCV, especially in resource-challenged regions.


Asunto(s)
Técnicas Biosensibles , Hepatitis C , Nanopartículas del Metal , Humanos , Hepacivirus/genética , Oro , Sensibilidad y Especificidad , Regiones no Traducidas 5' , Hepatitis C/diagnóstico , Técnicas de Amplificación de Ácido Nucleico/métodos , Técnicas de Diagnóstico Molecular/métodos , Técnicas Biosensibles/métodos
4.
Cytotherapy ; 2024 May 31.
Artículo en Inglés | MEDLINE | ID: mdl-38934983

RESUMEN

BACKGROUND AIMS: With the continuous development and advancement of human pluripotent stem cell (PSC)-derived cell therapies, an ever-increasing number of clinical indications can benefit from their application. Due to the capacity for PSCs to form teratomas, safety testing is required to ensure the absence of residual PSCs in a cell product. To mitigate these limitations, in vitro analytical methods can be utilized as quality control after the production of a PSC-derived cell product. Sensitivity of these analytic methods is critical in accurately quantifying residual PSC in the final cell product. In this study, we compared the sensitivity of three in vitro assays: qPCR, ddPCR and RT-LAMP. METHODS: The spike-in samples were produced from three independent experiments, each spiked with different PSC lines (PSC1, NH50191, and WA09 referred to as H9) into a background of primary fibroblasts (Hs68). These samples were then subjected to qPCR, ddPCR and RT-LAMP to determine their detection limit in measuring a commonly used PSC marker, LIN28A. RESULTS: The results indicated that the three analytic methods all exhibited consistent results across different cell-line spiked samples, with ddPCR demonstrating the highest sensitivity of the three methods. The LIN28A ddPCR assay could confidently detect 10 residual PSCs in a million fibroblasts. DISCUSSION: In our hand, ddPCR LIN28A assay demonstrated the highest sensitivity for detection of residual PSCs compared to the other two assays. Correlating such in vitro safety results with corresponding in vivo studies demonstrating the tumorigenicity profile of PSC-derived cell therapy could accelerate the safe clinical translation of cell therapy.

5.
Eur J Clin Microbiol Infect Dis ; 43(8): 1597-1607, 2024 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-38833104

RESUMEN

PURPOSE: To comprehensively investigate the diagnostic performance of routinely used assays in MPXV testing, the National Center of Clinical Laboratories in China conducted a nationwide external quality assessment (EQA) scheme and an evaluated nine assays used by ≥ 5 laboratories in the EQA. METHODS: MPXV virus-like particles with 2700, 900 and 300 copies/mL were distributed to 195 EQA laboratories. For extended analysis, triple-diluted samples from 9000 to 4.12 copies/mL were repeated 20 times using the assays employed by ≥ 5 laboratories. The diagnostic performance was assessed by analyzing EQA data and calculating the limits of detection (LODs). RESULTS: The performance was competent in 87.69% (171/195) of the participants and 87.94% (175/199) of the datasets. The positive percentage agreements (PPAs) were greater than 99% for samples at 2700 and 900 copies/mL, and 95.60% (761/796) for samples at 300 copies/mL. The calculated LODs for the two clades ranged from 228.44 to 924.31 copies/mL and were greater than the LODs specified by the respective kits. EasyDiagnosis had the lowest calculated LODs and showed superior performance in EQA, whereas BioGerm and Sansure, with higher calculated LODs, did not perform well in EQA. CONCLUSION: This study provides valuable information from the EQA data and evaluation of the diagnostic performance of MPXV detection assays. It also provided insights into reagent optimization and enabled prompt public health interventions for the outbreak.


Asunto(s)
Reacción en Cadena en Tiempo Real de la Polimerasa , Sensibilidad y Especificidad , Humanos , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , China/epidemiología , Límite de Detección , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Diagnóstico Molecular/normas , Monkeypox virus/genética , Monkeypox virus/aislamiento & purificación
6.
Med Mycol ; 62(2)2024 Jan 27.
Artículo en Inglés | MEDLINE | ID: mdl-38327232

RESUMEN

Mucormycosis is a rare disease with scarce diagnostic methods for early intervention. Available strategies employing direct microscopy using calcofluor white-KOH, culture, radiologic, and histopathologic testing often are time-intensive and demand intricate protocols. Nucleic Acid Amplification Test holds promise due to its high sensitivity combined with rapid detection. Loop-mediated isothermal amplification (LAMP) based detection offers an ultrasensitive technique that does not require complicated thermocyclers like in polymerase chain reaction, offering a straightforward means for improving diagnoses as a near-point-of-care test. The study introduces a novel magnetic nanoparticle-based LAMP assay for carryover contaminant capture to reduce false positives. Solving the main drawback of LAMP-based diagnosis techniques. The assay targets the cotH gene, which is invariably specific to Mucorales. The assay was tested with various species of Mucorales, and the limit of detections for Rhizopus microsporus, Lichtheimia corymbifera, Rhizopus arrhizus, Rhizopus homothallicus, and Cunninghamella bertholletiae were 1 fg, 1 fg, 0.1 pg, 0.1 pg, and 0.01 ng, respectively. This was followed by a clinical blindfolded study using whole blood and urine samples from 30 patients diagnosed with Mucormycosis. The assay has a high degree of repeatability and had an overall sensitivity of > 83%. Early Mucormycosis detection is crucial, as current lab tests from blood and urine lack sensitivity and take days for confirmation despite rapid progression and severe complications. Our developed technique enables the confirmation of Mucormycosis infection in < 45 min, focusing specifically on the RT-LAMP process. Consequently, this research offers a viable technique for quickly identifying Mucormycosis from isolated DNA of blood and urine samples instead of invasive tissue samples.


Mucormycosis is a challenging disease to diagnose early. This study introduces a sensitive and rapid diagnostic approach using Loop-mediated isothermal amplification technology. Testing blood and urine samples from 30 patients revealed promising sensitivity and repeatability, indicating its potential for non-invasive diagnosis.


Asunto(s)
Nanopartículas de Magnetita , Mucorales , Mucormicosis , Humanos , Mucormicosis/diagnóstico , Mucormicosis/veterinaria , Sensibilidad y Especificidad , Técnicas de Amplificación de Ácido Nucleico/métodos , Técnicas de Amplificación de Ácido Nucleico/veterinaria , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Diagnóstico Molecular/veterinaria , Mucorales/genética
7.
J Fluoresc ; 2024 Mar 04.
Artículo en Inglés | MEDLINE | ID: mdl-38436821

RESUMEN

In this review paper we discussed the detection of melatonin and serotonin by using various optical methods. Melatonin and serotonin are very necessary body hormones these are also called neuroregulatory hormones secreted by pineal gland in brain by pinealocytes and shape of pineal gland is cone like. Sensitive detection of melatonin and serotonin in pharmacological samples and human serum is crucial for human beings, lots of research publications available in literature for melatonin and serotonin and we overviewed these papers. We have deeply reviewed many research papers where sensitively sensing of melatonin and serotonin occurs, by using of various interfering agents and nanomaterials. This review aims presenting colorimetry, fluorometry and spectrophotometric detection of melatonin (MEL) and serotonin (SER) by using different metal oxides, carbon nanomaterials (nanosheets, nanorods, nanofibers) and many other agents. Nanomaterials typically possess favourable optical, electrical and mechanical characteristics, they provide up new avenues for enhancing the efficacy of sensors. It is crucial to provide an optical sensors platform that is dependable, sensitive and low price. The development of sensors and biosensors to use nanomaterials for neurotransmitters has advanced significantly in recent years. There are currently many developing biomarkers in biological fluids, and bionanomaterial-based biosensor systems, as well as clinical and pharmacological settings, have garnered significant interest. Biomarkers have been found using optical devices in a quick, selective and sensitive manner. Our aim is to compile all the data that already published on MEL, SER sensing and comparison of each method, we mainly focused on principle, observations, sensitivity, selectivity, limit of detection, mechanism behind the reaction, effect of temperature, pH and concentration. In the last of this paper, we discuss some challenges of these methods and future projects.

8.
Avian Pathol ; 53(2): 101-105, 2024 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-38018364

RESUMEN

RESEARCH HIGHLIGHTS: A sandwich ELISA was developed to detect EDSV using the mAbs 5G4 and HRP-6G6.The sandwich ELISA maintained high specificity and sensitivity.The sandwich ELISA had equivalent consistency with real-time PCR assay.


Asunto(s)
Anticuerpos Monoclonales , Atadenovirus , Animales , Ensayo de Inmunoadsorción Enzimática/veterinaria , Sensibilidad y Especificidad
9.
Mikrochim Acta ; 191(4): 183, 2024 Mar 07.
Artículo en Inglés | MEDLINE | ID: mdl-38451315

RESUMEN

A tunable plasmonic sensor has been developed by varying the dextran content in the initially synthesized dextran-gold nanoparticle (dAuNPs) solution. A colloidal nanogold solution (dAuNPs-Sol) was initially prepared using dextran and gold salt in alkaline media by a one-pot green synthetic route. The dAuNPs-Sol was combined with varying amounts of dextran (ranging from 0.01 to 30.01%) to create a tunable probe, along with different solid formats, including tablet (dAuNPs-Tab), powder (dAuNPs-Powder), and composite (dAuNPs-Comp). Both the liquid and solid phase plasmonic probes were characterized using UV-vis spectroscopy, transmission electron microscopy (TEM) dynamic light scattering (DLS), and zeta potential analysis. The impact of dextran content in the dAuNP solution is studied in terms of surface charge and hydrodynamic size. The influence of operational treatments used to achieve solid dAuNPs probes is also explored. All plasmonic probes were employed to detect a broad range of OCl¯ concentrations (ranging from µM to mM) in water through aggregation followed by calculating a lower and upper limit of detection (LLoD, ULoD) of the proposed colorimetric sensors. Results indicate that the most sensitive detection is achieved with a lower dextran content (0.01%), which exhibits an LLoD of 50 µM. The dAuNPs-Sol sensor is selective and demonstrates real-world applicability, as confirmed by interference analysis and successful testing with various water samples. Additionally, it is found that a 20 × concentration of dextran-coated gold nanoparticles could be attained without any changes in the particle morphology. This concentration is achieved through a straightforward process that does not require the use of a centrifuge machine. This finding highlights the practicality and simplicity of the method, indicating its potential for scalable and cost-effective production of concentrated dAuNPs without compromising their structural integrity.

10.
Sensors (Basel) ; 24(11)2024 Jun 04.
Artículo en Inglés | MEDLINE | ID: mdl-38894440

RESUMEN

Quasi-bound state in the continuum (QBIC) can effectively enhance the interaction of terahertz (THz) wave with matter due to the tunable high-Q property, which has a strong potential application in the detection of low-concentration biological samples in the THz band. In this paper, a novel THz metamaterial sensor with a double-chain-separated resonant cavity structure based on QBIC is designed and fabricated. The process of excitation of the QBIC mode is verified and the structural parameters are optimized after considering the ohmic loss by simulations. The simulated refractive index sensitivity of the sensor is up to 544 GHz/RIU, much higher than those of recently reported THz metamaterial sensors. The sensitivity of the proposed metamaterial sensor is confirmed in an experiment by detecting low-concentration lithium citrate (LC) and bovine serum albumin (BSA) solutions. The limits of detection (LoDs) are obtained to be 0.0025 mg/mL (12 µM) for LC and 0.03125 mg/mL (0.47 µM) for BSA, respectively, both of which excel over most of the reported results in previous studies. These results indicate that the proposed THz metamaterial sensor has excellent sensing performances and can well be applied to the detection of low-concentration biological samples.


Asunto(s)
Técnicas Biosensibles , Albúmina Sérica Bovina , Albúmina Sérica Bovina/química , Técnicas Biosensibles/métodos , Técnicas Biosensibles/instrumentación , Límite de Detección , Animales , Radiación Terahertz , Bovinos , Espectroscopía de Terahertz/métodos , Refractometría , Compuestos de Litio/química , Ácido Cítrico/química
11.
Sensors (Basel) ; 24(5)2024 Feb 24.
Artículo en Inglés | MEDLINE | ID: mdl-38475005

RESUMEN

In this work, we present a compact LIBS sensor developed for characterization of samples on a crime scene following requirements of law enforcement agencies involved in the project. The sensor operates both in a tabletop mode, for aside measurements of swabbed materials or taken fragments, and in handheld mode where the sensor head is pointed directly on targets at the scene. The sensor head is connected via an umbilical to an instrument box that could be battery-powered and contains also a color camera for sample visualization, illumination LEDs, and pointing system for placing the target in focus. Here we describe the sensor's architecture and functionalities, the optimization of the acquisition parameters, and the results of some LIBS measurements. On nano-plotted traces at silica wafer and in optimized conditions, for most of the elements the detection limits, in term of the absolute element masses, were found to be below 10 picograms. We also show results obtained on some representative materials, like fingerprints, swabbed soil and gunshot residue, varnishes on metal, and coated plastics. The last, solid samples were used to evaluate the depth profiling capabilities of the instrument, where the recognition of all four car paint layers was achieved.

12.
J Aquat Anim Health ; 2024 Apr 15.
Artículo en Inglés | MEDLINE | ID: mdl-38622805

RESUMEN

OBJECTIVE: We sought to evaluate accurate and reproducible detection of Myxobolus cerebralis (Mc), the causative agent of whirling disease, by using nested polymerase chain reaction (nPCR) and three previously established real-time quantitative PCR (qPCR) assays: K18S (Kelley 18S), C18S (Cavender 18S), and Hsp70 (heat shock protein 70). We used a "fit for purpose" approach combined with intra- and interlaboratory testing to identify a molecular testing method that would be equivalent to the currently accepted nPCR procedure for Mc. METHODS: Assay performance was compared using a combination of intra- and interlaboratory testing that used synthetic gBlocks along with naturally and experimentally infected fish tissue. North American isolates representing geographically distinct locations were also tested using all three assays. RESULT: The K18S and C18S assays exhibited high assay sensitivity, intra- and interlaboratory repeatability of sample replicates, and reproducible identification of all test samples across multiple laboratories. In contrast, the Hsp70 assay failed to detect several positive samples at low DNA concentrations during intra- and interlaboratory testing. The K18S assay was the only procedure that demonstrated perfect detection accuracy when testing geographically distinct Mc isolates. Results demonstrated the K18S assay is robust under variable test conditions, is more accurate than the C18S and Hsp70 assays, and provides detection capabilities equivalent to those of the currently accepted nPCR confirmation assay "gold standard" that is described in the American Fisheries Society-Fish Health Section (AFS-FHS) Blue Book. CONCLUSION: The "fit for purpose" approach and preliminary completion of the World Organization for Animal Health validation pathway demonstrate that the K18S assay provides an alternate method for Mc testing. This work provides the foundation for acceptance of the K18S assay into the AFS-FHS Blue Book as a standardized test procedure for Mc.

13.
Behav Res Methods ; 56(3): 1207-1228, 2024 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-38129736

RESUMEN

Data censoring occurs when researchers do not know precise values of data points (e.g., age is 55+ or concentration ≤ .001). Censoring is frequent within psychology but typically unrecognized outside of longitudinal studies. We describe five circumstances when censoring may occur, demonstrate censoring distorts correlations, and discuss how censoring can create spurious factors. Next, we explain how to use R package lava to calculate maximum likelihood estimates (Holst and Budtz-Jørgensen Computational Statistics, 28(4), 1385-1452, 2013) of correlations between uncensored variables based upon censored variables. Previous research demonstrated these estimates were more accurate than Muthén's (1984) estimate for one particular model, but no research has systematically examined their accuracy. We therefore conducted a simulation study exploring the effects of the correlation, sample size, and censoring on point and interval estimates of correlations. Based upon 80 cells in which low values of normally distributed variables were censored, we recommend the constrained regression model with Wald confidence intervals. These methods were precise and unbiased unless both variables had 70% censoring and the correlation was large and negative (e.g., -.9), in which case estimates were closer to -1 than they should be. Opposite results would occur if low values of one variable and high values of the other were censored: Estimates would be precise and unbiased unless censoring was extreme and correlations were large and positive. To estimate large correlations accurately, we recommend researchers reduce censoring by using longer longitudinal studies, using scales with more response options, and matching measures to populations to reduce floor and ceiling effects.


Asunto(s)
Simulación por Computador , Humanos , Funciones de Verosimilitud , Estudios Longitudinales , Interpretación Estadística de Datos
14.
J Clin Microbiol ; 61(8): e0051823, 2023 08 23.
Artículo en Inglés | MEDLINE | ID: mdl-37432105

RESUMEN

The measurement of Epstein-Barr virus (EBV) deoxyribonucleic acid (DNA) is key to diagnosing and managing EBV-associated complications in transplant recipients. The performance of the new Conformité Européenne (CE) and Food and Drug Administration (FDA)-cleared quantitative Roche cobas EBV real-time PCR assay was determined by using EDTA-plasma dilution panels and clinical samples that were spiked with either the World Health Organization's EBV international standard or high-titer EBV lambda stock. Correlation with the Abbott Realtime EBV assay was assessed in clinical specimens and conducted at two independent laboratories. An in silico analysis revealed that the dual-target test (EBNA1 and BMRF2) was 100% inclusive for the known diversity of EBV. The overall limit of detection (LoD) was 16.6 IU/mL for genotype 1 (GT1). GT2 LoD was verified at 18.8 IU/mL. The linear ranges were from 1.40 × 101 to 2.30 × 108 IU/mL and from 2.97 × 101 to 9.90 × 107 IU/mL for GT1 and GT2, respectively. Accuracy was confirmed across the linear range (mean difference not exceeding ±0.18 log10). Precision was not influenced by the factors analyzed (standard deviation of 0.02 to 0.17 log10), including the presence of potentially interfering endogenous or exogenous substances. Plasma samples were stable under several conditions (variable time points, storage, and freeze/thaw cycles). In clinical EBV DNA-positive samples, correlation between the cobas EBV test and the comparator was high (n = 126 valid results; R2 = 0.96) with a 0.1 mean log10 titer difference. The cobas EBV test is an accurate, sensitive, specific, and reproducible assay for the detection of EBV DNAemia in plasma. In general, high levels of automation and calibration to the international standard will lead to improvements in the harmonization of quantitative EBV DNA test results across laboratories.


Asunto(s)
Infecciones por Virus de Epstein-Barr , Herpesvirus Humano 4 , Humanos , Herpesvirus Humano 4/genética , Infecciones por Virus de Epstein-Barr/diagnóstico , Sensibilidad y Especificidad , Carga Viral/métodos , ADN , Técnicas de Diagnóstico Molecular/métodos , ADN Viral/genética
15.
J Med Virol ; 95(1): e28395, 2023 01.
Artículo en Inglés | MEDLINE | ID: mdl-36504122

RESUMEN

Rapid and accurate diagnosis of infections is fundamental to containment of disease. Several monkeypox virus (MPV) real-time diagnostic assays have been recommended by the CDC; however, the specificity of the primers and probes in these assays for the ongoing MPV outbreak has not been investigated. We analyzed the primer and probe sequences present in the CDC recommended MPV generic real-time PCR assay by aligning those sequences against 1730 MPV complete genomes reported in 2022 worldwide. Sequence mismatches were found in 99.08% and 97.46% of genomes for the MPV generic forward and reverse primers, respectively. Mismatch-corrected primers were synthetized and compared to the generic assay for MPV detection. Results showed that the two primer-template mismatches resulted in a ~11-fold underestimation of initial template DNA in the reaction and 4-fold increase in the 95% LOD. We further evaluated the specificity of seven other real-time PCR assays used for MPV and orthopoxvirus (OPV) detection and identified two assays with the highest matching score (>99.6%) to the global MPV genome database in 2022. Genetic variations in the primer-probe regions across MPV genomes could indicate the temporal and spatial emergence pattern of monkeypox disease. Our results show that the current MPV real-time generic assay may not be optimal to accurately detect MPV, and the mismatch-corrected assay with full complementarity between primers and current MPV genomes could provide a more sensitive and accurate detection of MPV.


Asunto(s)
Monkeypox virus , Mpox , Humanos , Monkeypox virus/genética , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Cartilla de ADN/genética , Mpox/diagnóstico , Mpox/epidemiología , Brotes de Enfermedades , Sensibilidad y Especificidad
16.
Stat Med ; 42(2): 164-177, 2023 01 30.
Artículo en Inglés | MEDLINE | ID: mdl-36404417

RESUMEN

In vaccine research towards the prevention of infectious diseases, immune response biomarkers serve as an important tool for comparing and ranking vaccine candidates based on their immunogenicity and predicted protective effect. However, analyses of immune response outcomes can be complicated by differences across assays when immune response data are acquired from multiple groups/laboratories. Motivated by a real-world problem to accommodate the use of two different neutralization assays in COVID-19 vaccine trials, we propose methods based on left-censored multivariate normal model assuming common assay differences across settings, to adjust for differences between assays with respect to measurement error and the lower limit of detection. Our proposed methods integrate external paired-sample data with bridging assumptions to achieve two objectives, both using pooled data acquired from different assays: (i) comparing immunogenicity between vaccine regimens, and (ii) evaluating correlates of risk. In simulation studies, for the first objective, our method leads to unbiased calibrated assay mean with good coverage of bootstrap confidence interval, as well as valid test for immunogenicity comparison, while the alternative method assuming constant calibration model between assays leads to biased estimate of assay mean with undercoverage problem and invalid test with inflated type-I error; for the second objective, in the presence of noticeable left-censoring rate, our proposed method can drastically outperform the existing method that ignores left-censoring, in terms of reduced bias and improved precision. We apply the proposed methods to SARS-CoV-2 spike-pseudotyped virus neutralization assay data generated in vaccine and convalescent samples by two different laboratories.


Asunto(s)
COVID-19 , Vacunas , Humanos , SARS-CoV-2 , COVID-19/prevención & control , Vacunas contra la COVID-19 , Simulación por Computador , Anticuerpos Antivirales
17.
Stat Med ; 42(2): 193-208, 2023 01 30.
Artículo en Inglés | MEDLINE | ID: mdl-36457137

RESUMEN

Understanding the association between mixtures of environmental toxicants and time-to-pregnancy (TTP) is an important scientific question as sufficient evidence has emerged about the impact of individual toxicants on reproductive health and that individuals are exposed to a whole host of toxicants rather than an individual toxicant. Assessing mixtures of chemical effects on TTP poses significant statistical challenges, namely (i) TTP being a discrete survival outcome, typically subject to left truncation and right censoring, (ii) chemical exposures being strongly correlated, (iii) appropriate transformation to account for some lipid-binding chemicals, (iv) non-linear effects of some chemicals, and (v) high percentage of concentration below the limit of detection (LOD) for some chemicals. We propose a discrete frailty modeling framework (named Discnet) that allows selection of correlated covariates while appropriately addressing the methodological issues mentioned above. Discnet is shown to have better and stable false negative and false positive rates compared to alternative methods in various simulation settings. We did a detailed analysis of the pre-conception endocrine disrupting chemicals and TTP from the LIFE study and found that older females, female exposure to cotinine (smoking), DDT conferred a delay in getting pregnant, which was consistent across various approaches to account for LOD as well as non-linear associations.


Asunto(s)
Fragilidad , Tiempo para Quedar Embarazada , Embarazo , Humanos , Femenino , Sustancias Peligrosas , Simulación por Computador , Límite de Detección
18.
Photochem Photobiol Sci ; 22(11): 2483-2497, 2023 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-37747667

RESUMEN

The present paper reports on the synthesis and characterization of a new chemosensor for fluoride ions, a hydrazone derived from pyridoxal 5'-phosphate and benzothiazole. The structure of the chemosensor was confirmed using 1H and 13C NMR, FT-IR and mass spectroscopy. The conformational diversity of the chemosensor influencing the sensor activity was studied by the quantum chemistry methods on the B3LYP/6-311++G(d, p) (H, C, N, O, P, S) level, and the optimal structure of the chemosensor was chosen. The selective capability of detecting F- in the aqueous solution, which also contains Cl-, Br-, I-, NCS-, ClO4-, HSO4-, and NO3- was demonstrated. The detection limit (LOD) for fluoride ions was 0.22 µM as determined by the 3σ method. The turn-on effect in the presence of fluoride ions is based on the deprotonation of the chemosensor and its subsequent aggregation in DMSO. In addition, the chemosensor was used for the detection and estimation of F- in real samples using fluorescence spectroscopy.

19.
J Fluoresc ; 2023 Dec 01.
Artículo en Inglés | MEDLINE | ID: mdl-38038876

RESUMEN

The chemosensors act as powerful tool in the detection of metal ions due to their simplicity, high sensitivity, low cost, low detection limit, rapid photophysical response, and application to the environmental and medical fields. This review article presents an overview for the chemosensing of Ag+ ions based on Calix, MOF, Nanoparticle, COF, Calix, Electrochemical chemosensor published from 2018 to 2023. Here, we have reviewed the sensing of Ag+ ions and summarised the binding response, mechanism, LOD, colorimetric response, adsorption capacity, technique used. The purpose of this review article to provide a detailed summary of the performance of different host chemosensors that are helpful for providing future direction to researchers on Ag+ ion detection and provides path to design effective chemsosensor (simple to synthesize, cost effective, high sensitivity, with more practical application). While studying the related article literature, we came across some challenges and that has been discussed lastly and provided solutions for them.

20.
Sens Actuators B Chem ; 392: 134085, 2023 Oct 01.
Artículo en Inglés | MEDLINE | ID: mdl-37304211

RESUMEN

Sensitive and rapid detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been a vital goal in the ongoing COVID-19 pandemic. We present in this comprehensive work, for the first time, detailed fabrication and clinical validation of a point of care (PoC) device for rapid, onsite detection of SARS-CoV-2 using a real-time reverse-transcription loop-mediated isothermal amplification (RT-LAMP) reaction on a polymer cartridge. The PoC system, namely PATHPOD, consisting of a standalone device (weight less than 1.2 kg) and a cartridge, can perform the detection of 10 different samples and two controls in less than 50 min, which is much more rapid than the golden standard real-time reverse-transcription Polymerase Chain Reaction (RT-PCR), typically taking 16-48 h. The novel total internal reflection (TIR) scheme and the reactions inside the cartridge in the PoC device allow monitoring of the diagnostic results in real-time and onsite. The analytical sensitivity and specificity of the PoC test are comparable with the current RT-PCR, with a limit of detection (LOD) down to 30-50 viral genome copies. The robustness of the PATHPOD PoC system has been confirmed by analyzing 398 clinical samples initially examined in two hospitals in Denmark. The clinical sensitivity and specificity of these tests are discussed.

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