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1.
Chembiochem ; 25(3): e202300575, 2024 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-37963820

RESUMO

Salmonella constitutes a prevalent alimentary pathogen, instigating zoonotic afflictions. Consequently, the prompt discernment of Salmonella in sustenance is of cardinal significance. Lateral flow assays utilizing colorimetric methodologies adequately fulfill the prerequisites of point-of-care diagnostics, however, their detection threshold remains elevated, generally permitting only qualitative discernment, an impediment to the preliminary screening of nascent pathogens. In response to this conundrum, we propose a lateral flow diagnostic predicated upon a streptavidin-biotin amplification system with recombinase polymerase amplification engineered for the expeditious and quantitative discernment of Salmonella enteritidis. Trace nucleic acids within a sample undergo exponential amplification via recombinase polymerase amplification to a level discernable, constituting the initial signal amplification. Subsequently, along the test line (T-line) of the lateral flow strip, the chromatic signal undergoes augmentation by securing a greater quantity of AuNPs through the magnification capacity of the streptavidin-biotin mechanism, affecting the second signal amplification. Quantitative results are procured via smartphone capture and transferred to computer software for precise calculation of the targeted quantity. The lateral flow strip exhibits a LOD at 19.41 CFU/mL for cultured S. enteritidis. The RSD of three varying concentrations were respectively 3.74 %, 5.96 %, and 4.25 %.


Assuntos
Nanopartículas Metálicas , Salmonella enteritidis , Salmonella enteritidis/genética , Biotina , Estreptavidina , Recombinases , Ouro , Nucleotidiltransferases , Técnicas de Amplificação de Ácido Nucleico/métodos , Sensibilidade e Especificidade
2.
Eur J Clin Microbiol Infect Dis ; 43(9): 1815-1823, 2024 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-39012550

RESUMO

PURPOSE: This study aimed to develop a double antigen sandwich ELISA (DAgS-ELISA) method for more efficient, accurate, and quantitative detection of total antibodies against Candida albicans enolase1 (CaEno1) for diagnosing invasive candidiasis (IC). METHODS: DAgS-ELISA was developed using recombinant CaEno1 and a monoclonal antibody as the standard. Performance evaluation included limit of detection, accuracy, and repeatability. Dynamic changes in antibody levels against CaEno1 in serum from systemic candidiasis mice were analyzed using DAgS-ELISA. Patient serum samples from IC, Candida colonization, bacterial infections, and healthy controls were analyzed with DAgS-ELISA and indirect ELISA. RESULTS: DAgS-ELISA outperformed indirect ELISA in terms of linear range and test background. In systemic candidiasis mice, a distinctive 'double-peak' pattern in dynamic antibody levels was observed. Additionally, there was a high level of consistency in the positive rates of CaEno1 antibodies detected by both DAgS-ELISA and indirect ELISA. While the positivity rates differed among patient groups, no significant variations in antibody levels were detected among the various positive patient groups. CONCLUSIONS: DAgS-ELISA offers a reliable novel approach for IC diagnosis, enabling rapid, accurate, and quantitative detection of CaEno1 antibodies. Further validation and optimization are needed for its clinical application and effectiveness.


Assuntos
Anticorpos Antifúngicos , Candida albicans , Ensaio de Imunoadsorção Enzimática , Fosfopiruvato Hidratase , Ensaio de Imunoadsorção Enzimática/métodos , Animais , Fosfopiruvato Hidratase/imunologia , Fosfopiruvato Hidratase/sangue , Candida albicans/imunologia , Anticorpos Antifúngicos/sangue , Camundongos , Humanos , Candidíase Invasiva/diagnóstico , Candidíase Invasiva/imunologia , Candidíase Invasiva/sangue , Feminino , Candidíase/diagnóstico , Candidíase/sangue , Candidíase/imunologia , Antígenos de Fungos/imunologia , Antígenos de Fungos/sangue , Sensibilidade e Especificidade , Proteínas Fúngicas/imunologia , Anticorpos Monoclonais/imunologia , Camundongos Endogâmicos BALB C
3.
Nanotechnology ; 35(17)2024 Feb 09.
Artigo em Inglês | MEDLINE | ID: mdl-38271716

RESUMO

A visual detection method for malachite green (MG) in food was established based on 'double-response-OFF' ratiometric fluorescent paper-based sensor. Biomass carbon quantum dots (BCQDs) using broad bean shell, and Ti3C2MXene quantum (MQDs) dots modified by ethylenediamine were synthesized by solvothermal method. The MG and two kinds of quantum dots could undergo static quenching, and the fluorescence color of two kinds of quantum dots gradually changed from red to blue, eventually the fluorescence was quenched, and the pattern had a two-stage linear relationship using fluorescent spectrofluorometer in the range of 0.1-140.0µM and the detection limit of 0.07µM. On this basis, a BCQDs/MQDs ratiometric fluorescence paper-based sensor was constructed and applied to fish sample. Through mobile phone software-Color recognizer, RGB values of fluorescent paper-based sensor at various concentrations of MG were extracted. The results showed that MG concentration was linearly correlated withR' value of RGB in the range of 20.0-140.0µM with 16.5µM detection limit. The method had been applied to the determination of canned fish and fresh basa fish samples, and the recovery rates were 97.33%-108.93% and 96.04%-117.97%, respectively. It proved that the ratiometric fluorescent paper-based sensor could be used for the rapid visual quantitative detecting MG in real samples.


Assuntos
Nitritos , Pontos Quânticos , Corantes de Rosanilina , Elementos de Transição , Animais , Corantes Fluorescentes , Carbono , Titânio , Biomassa , Limite de Detecção , Espectrometria de Fluorescência , Peixes
4.
J Fluoresc ; 2024 Sep 05.
Artigo em Inglês | MEDLINE | ID: mdl-39235654

RESUMO

We aim to develop an amplified luminescence proximity homogeneous assay (AlphaLISA) for quantification of trypsinogen-2 levels in human serum for the diagnosis of acute pancreatitis. Based on new amplified luminescence proximity homogeneity assay (AlphaLISA) method, carboxyl-modified donor and acceptor beads were coupled to capture and detection antibodies. A double antibody sandwich immunoassay was used to detect the concentration of trypsinogen-2 in serum. The method had good linearity (> 0.998). The intra - analysis precision was between 1.54% and 2.20% (< 10%), the inter-analysis precision was between 3.17% and 6.94% (< 15%), and the recovery was between 96.23% and 103.45%. The cross-reactivity of carbohydrate antigen 242 (CA242) and T-cell immunoglobulin mucin-3 (Tim-3) were 0.09% and 0.93%, respectively. The detection time only needed 15 min. The results of trypsinogen-2-AlphaLISA and time-resolved fluorescence immunoassay were consistent (ρ = 0.9019). In addition, serum trypsinogen-2 concentration in patients with acute pancreatitis [239.23 (17.83-807.58) ng/mL] was significantly higher than that in healthy controls [20.54 (12.10-39.73) ng/mL]. When the cut-off value was 35.38ng/mL, the sensitivity and specificity were 91.8% and 96.67%, and the positive detection rate was 91.80%. We have successfully established a trypsinogen-2-AlphaLISA method, which can promote the timely diagnosis of acute pancreatitis.

5.
J Fluoresc ; 2024 Jun 12.
Artigo em Inglês | MEDLINE | ID: mdl-38865061

RESUMO

In order to design organic small molecule fluorescent materials with multiple sensing, a bibranched -NH2 modified cyanostilbene derivative (AM) was synthesized. It exhibits solvent and aggregation-induced emission effects, with a solid-state quantum yield of 28%, which is seven times higher than that in THF. The synthesized sample AM demonstrated high sensitivity to trace water via a fluorescence "turn-off" response, achieving a low detection limit of 0.41 µM in THF and 0.80 µM in EtOH. AM also exhibits a "turn-off" response to picric acid, attributed to the photo-induced electron transfer effect it induces. The recognition of picric acid by AM demonstrates specificity and resistance to interference from nitro explosives, with a detection limit of 300 ppb and a linear relationship (R2 = 0.9981) at the range of 0-4 equivalents AM. Such acid recognition can facilitate the design of qualitative test papers and safety inks. Additionally, AM can function as a temperature sensor with a linear relationship (R2 = 0.9976) within the temperature range of 25-110 °C. Leveraging these unique characteristics, a series of methods were proposed for the direct quantitative determination of trace water in nonaqueous solvents, picric acid, and temperature.

6.
Environ Res ; 249: 118466, 2024 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-38354882

RESUMO

Global outbreaks and the spread of viral diseases in the recent years have led to a rapid increase in the usage of antiviral drugs (ATVs), the residues and metabolites of which are discharged into the natural environment, posing a serious threat to human health. There is an urgent need to develop sensitive and rapid detection tools for multiple ATVs. In this study, we developed a highly sensitive electrochemical sensor comprising a glassy carbon electrode (GCE) modified with graphitized hydroxylated multi-walled carbon nanotubes (G-MWCNT-OH) and 1-butyl-3-methylimidazolium hexafluorophosphate (BMIMPF6, IL) for the detection of six ATVs including famciclovir (FCV), remdesivir (REM), favipiravir (FAV), hydroxychloroquine sulfate (HCQ), cepharanthine (CEP) and molnupiravir (MOL). The morphology and structure of the G-MWCNT-OH/IL nanocomposites were characterized comprehensively, and the electroactive surface area and electron conductivity of G-MWCNT-OH/IL/GCE were determined using cyclic voltammetry and electrochemical impedance spectroscopy. The thermodynamic stability and non-covalent interactions between the G-MWCNT-OH and IL were evaluated through quantum chemical simulation calculations, and the mechanism of ATV detection using the G-MWCNT-OH/IL/GCE was thoroughly examined. The detection conditions were optimized to improve the sensitivity and stability of electrochemical sensors. Under the optimal experimental conditions, the G-MWCNT-OH/IL/GCE exhibited excellent electrocatalytic performance and detected the ATVs over a wide concentration range (0.01-120 µM). The limit of detections (LODs) were 42.3 nM, 55.4 nM, 21.9 nM, 15.6 nM, 10.6 nM, and 3.2 nM for FCV, REM, FAV, HCQ, CEP, and MOL, respectively. G-MWCNT-OH/IL/GCE was also highly stable and selective to the ATVs in the presence of multiple interfering analytes. This sensor exhibited great potential for enabling the quantitative detection of multiple ATVs in actual water environment.


Assuntos
Antivirais , Técnicas Eletroquímicas , Líquidos Iônicos , Nanotubos de Carbono , Antivirais/análise , Antivirais/química , Nanotubos de Carbono/química , Líquidos Iônicos/química , Técnicas Eletroquímicas/instrumentação , Técnicas Eletroquímicas/métodos , Limite de Detecção , Eletrodos , Grafite/química
7.
Appl Microbiol Biotechnol ; 108(1): 45, 2024 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-38175238

RESUMO

Veillonella spp. are Gram-negative opportunistic pathogens present in the respiratory, digestive, and reproductive tracts of mammals. An abnormal increase in Veillonella relative abundance in the body is closely associated with periodontitis, inflammatory bowel disease, urinary tract infections, and many other diseases. We designed a pair of primers and a probe based on the 16S rRNA gene sequences of Veillonella and conducted real-time quantitative PCR (qPCR) and droplet digital PCR (ddPCR) to quantify the abundance of Veillonella in fecal samples. These two methods were tested for specificity and sensitivity using simulated clinical samples. The sensitivity of qPCR was 100 copies/µL, allowing for the accurate detection of a wide range of Veillonella concentrations from 103 to 108 CFU/mL. The sensitivity of ddPCR was 11.3 copies/µL, only allowing for the accurate detection of Veillonella concentrations from 101 to 104 CFU/mL because of the limited number of droplets generated by ddPCR. ddPCR is therefore more suitable for the detection of low-abundance Veillonella samples. To characterize the validity of the assay system, clinical samples from children with inflammatory bowel disease were collected and analyzed, and the results were verified using isolation methods. We conclude that molecular assays targeting the 16S rRNA gene provides an important tool for the rapid diagnosis of chronic and infectious diseases caused by Veillonella and also supports the isolation and identification of Veillonella for research purposes. KEY POINTS: • With suitable primer sets, the qPCR has a wider detection range than ddPCR. • ddPCR is suitable for the detection of low-abundance samples. • Methods successfully guided the isolation of Veillonella in clinical sample.


Assuntos
Doenças Inflamatórias Intestinais , Veillonella , Criança , Humanos , Bioensaio , Doenças Inflamatórias Intestinais/diagnóstico , Mamíferos , Reação em Cadeia da Polimerase em Tempo Real , RNA Ribossômico 16S/genética
8.
Sensors (Basel) ; 24(12)2024 Jun 12.
Artigo em Inglês | MEDLINE | ID: mdl-38931587

RESUMO

Track irregularities directly affect the quality and safety of railway vehicle operations. Quantitative detection and real-time monitoring of track irregularities are of great importance. However, due to the frequent variable vehicle speed, vehicle operation is a typical non-stationary process. The traditional signal analysis methods are unsuitable for non-stationary processes, making the quantitative detection of the wavelength and amplitude of track irregularities difficult. To solve the above problems, this paper proposes a quantitative detection method of track irregularities under non-stationary conditions with variable vehicle speed by order tracking analysis for the first time. Firstly, a simplified wheel-rail dynamic model is established to derive the quantitative relationship between the axle-box vertical vibration and the track vertical irregularities. Secondly, the Simpson double integration method is proposed to calculate the axle-box vertical displacement based on the axle-box vertical acceleration, and the process error is optimized. Thirdly, based on the order tracking analysis theory, the angular domain resampling is performed on the axle-box vertical displacement time-domain signal in combination with the wheel rotation speed signals, and the quantitative detection of the track irregularities is achieved. Finally, the proposed method is validated based on simulation and field test analysis cases. We provide theoretical support and method reference for the quantitative detection method of track irregularities.

9.
Molecules ; 29(12)2024 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-38931008

RESUMO

Flexible and transparent surface-enhanced Raman scattering (SERS) substrates have attracted considerable attention for their ability to enable the direct in situ detection of analytes on curved surfaces. However, the curvature of an object can impact the signal enhancement of SERS during the measurement process. Herein, we propose a simple approach for fabricating a curvature-insensitive transparent SERS substrate by depositing silver nanoparticles (Ag NPs) onto a large-area wrinkled polystyrene/polydimethylsiloxane (Ag NP@W-PS/PDMS) bilayer film. Using rhodamine 6G (R6G) as a probe molecule, the optimized Ag NP@W-PS/PDMS film demonstrates a high analytical enhancement factor (AEF) of 4.83 × 105, excellent uniformity (RSD = 7.85%) and reproducibility (RSD = 3.09%), as well as superior mechanical flexibility. Additionally, in situ measurements of malachite green (MG) on objects with diverse curvatures, including fish, apple, and blueberry, are conducted using a portable Raman system, revealing a consistent SERS enhancement. Furthermore, a robust linear relationship (R2 ≥ 0.990) between Raman intensity and the logarithmic concentration of MG detected from these objects is achieved. These results demonstrate the tremendous potential of the developed curvature-insensitive SERS substrate as a point-of-care testing (POCT) platform for identifying analytes on irregular objects.

10.
Annu Rev Phys Chem ; 73: 141-162, 2022 04 20.
Artigo em Inglês | MEDLINE | ID: mdl-34936809

RESUMO

Surface-enhanced Raman scattering (SERS), a powerful technique for trace molecular detection, depends on chemical and electromagnetic enhancements. While recent advances in instrumentation and substrate design have expanded the utility, reproducibility, and quantitative capabilities of SERS, some challenges persist. In this review, advances in quantitative SERS detection are discussed as they relate to intermolecular interactions, surface selection rules, and target molecule solubility and accessibility. After a brief introduction to Raman scattering and SERS, impacts of surface selection rules and enhancement mechanisms are discussed as they relate to the observation of activation and deactivation of normal Raman modes in SERS. Next, experimental conditions that can be used to tune molecular affinity to and density near SERS substrates are summarized and considered while tuning these parameters is conveyed. Finally, successful examples of quantitative SERS detection are discussed, and future opportunities are outlined.


Assuntos
Análise Espectral Raman , Reprodutibilidade dos Testes , Análise Espectral Raman/métodos
11.
J Fish Dis ; 46(4): 357-367, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36606558

RESUMO

Ichthyophthiriasis, caused by the parasitic ciliate Ichthyophthirius multifiliis (Ich), is considered one of the most harmful diseases affecting freshwater fish globally. It can cause mass mortalities of fish in intensive farming systems. In such systems, it is thus necessary to detect and quantify the number of Ich in the water so that control measures can be implemented before Ichthyophthiriasis breaks out. In recent years, molecular diagnostic methods have become increasingly important in aquaculture. Real-time quantitative polymerase chain reaction (qPCR) and droplet digital polymerase chain reaction (ddPCR) have become robust assays for detecting pathogens. In this study, a set of specific primers and a TaqMan-minor groove binder probe targeting the small-subunit rDNA (SSU rDNA) of Ich were developed. They were used in qPCR and ddPCR assays to compare the performance of these two different methods in quantitatively detecting Ich. After optimizing the reaction conditions, both qPCR and ddPCR assays were found to have high linearity and quantitative correlations for standard plasmid DNA. When used for the detection of Ich eDNA in water samples, the qPCR assay had a wider detection range, making it a suitable method to screen for the prevalence of Ichthyophthiriasis. However, the ddPCR approach had higher sensitivity, which would help provide advance notice of the disease in complex water environmental samples.


Assuntos
Doenças dos Peixes , Hymenostomatida , Animais , Doenças dos Peixes/diagnóstico , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Reação em Cadeia da Polimerase em Tempo Real/métodos , Água Doce , Água , DNA Ribossômico
12.
Mikrochim Acta ; 190(10): 392, 2023 09 15.
Artigo em Inglês | MEDLINE | ID: mdl-37713003

RESUMO

A novel and portable detection platform for procymidone (PRM) was developed by combining simple sample pretreatment, lateral flow test strips based on multi-branched gold nanoparticle (LFTS-MBGNP), and a smartphone. Based on the large surface area of MBGNPs, rapid detection of PRM was realized by simple naked eye observation. By utilizing a smartphone as a portable signal analyzer, ultrasensitive quantitative detection of PRM in red wine was realized with the limits of detection (LOD) of 1.60 ng/mL, which was 3000 times lower than the US limit (5 ppm). Moreover, rapid detection of four kinds of fruits and vegetables was achieved within 10 min, with LODs of 4.34 ng/g, 6.93 ng/g, 8.99 ng/g, and 5.03 ng/g, respectively, which could meet the PRM limit of the European Union (10 ng/g). Integrating the optimized QuEChERS pretreatment method, the developed platform realized a simple and sensitive on-site detection of PRM pesticide in foods and red wine within 45 min. This platform provides a useful tool and new idea for rapid screening and detection of pesticide residues in food.


Assuntos
Nanopartículas Metálicas , Resíduos de Praguicidas , Praguicidas , Ouro
13.
Sensors (Basel) ; 23(24)2023 Dec 11.
Artigo em Inglês | MEDLINE | ID: mdl-38139607

RESUMO

This paper introduces a novel method for enhancing underground pipeline inspection, specifically addressing limitations associated with traditional closed-circuit television (CCTV) systems. These systems, commonly used for capturing visual data of sewer system deformations, heavily rely on subjective human expertise, leading to limited accuracy in detection. Furthermore, their inability to perform quantitative analyses of deformation extent hampers overall inspection effectiveness. Our proposed method leverages laser point cloud data and employs a 3D scanner for objective detection of geometric deformations in underground pipe corridors. By utilizing this approach, we enable a quantitative assessment of blockage levels, offering a significant improvement over traditional CCTV-based methods. The key advantages of our method lie in its objectivity and quantification capabilities, ultimately enhancing detection reliability, accuracy, and overall inspection efficiency.

14.
Molecules ; 28(11)2023 May 24.
Artigo em Inglês | MEDLINE | ID: mdl-37298764

RESUMO

ReS2, as a new member of transition metal dichalcogenides (TMDCs), has emerged as a promising substrate for semiconductor surface-enhanced Raman spectroscopy (SERS) due to its unique optoelectronic properties. Nevertheless, the sensitivity of the ReS2 SERS substrate poses a significant challenge to its widespread application in trace detection. In this work, we present a reliable approach for constructing a novel ReS2/AuNPs SERS composite substrate, enabling ultrasensitive detection of trace amounts of organic pesticides. We demonstrate that the porous structures of ReS2 nanoflowers can effectively confine the growth of AuNPs. By precisely controlling the size and distribution of AuNPs, numerous efficient and densely packed "hot spots" were created on the surface of ReS2 nanoflowers. As a result of the synergistic enhancement of the chemical and electromagnetic mechanisms, the ReS2/AuNPs SERS substrate demonstrates high sensitivity, good reproducibility, and superior stability in detecting typical organic dyes such as rhodamine 6G and crystalline violet. The ReS2/AuNPs SERS substrate shows an ultralow detection limit of 10-10 M and a linear detection of organic pesticide molecules within 10-6-10-10 M, which is significantly lower than the EU Environmental Protection Agency regulation standards. The strategy of constructing ReS2/AuNPs composites would contribute to the development of highly sensitive and reliable SERS sensing platforms for food safety monitoring.


Assuntos
Nanopartículas Metálicas , Praguicidas , Ouro/química , Nanopartículas Metálicas/química , Reprodutibilidade dos Testes , Análise Espectral Raman/métodos
15.
Int Arch Allergy Immunol ; 183(5): 490-497, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-34915490

RESUMO

BACKGROUND: Light-initiated chemiluminescence assay (LICA) is a homogeneous assay that has been successfully used for the quantitation of food allergen-specific immunoglobulin E (sIgE), but not inhaled allergen-sIgE. Simultaneously, current assays used to detect allergen-sIgE are serum consuming and/or time consuming. Hence, we established a method for the quantitation of Artemisia-sIgE based on LICA and verified its performance according to the clinical guideline documents, laying a foundation for the quantitation of inhaled and food allergen-sIgE in parallel on LICA. METHODS: The assay was established after optimizing the first incubation time and the dilutions of Artemisia-coated chemibeads, biotinylated goat anti-human IgE, and serum. In order to quantitate Artemisia-sIgE, the calibration curve was established with a high positive serum of known concentration. The assay performance was confirmed per the clinical guideline documents. In addition, the correlation between the results of LICA and capture enzyme-linked immunosorbent assay was evaluated. RESULTS: The developed LICA's coefficients of variation of repeatability and intermediate precision were 3.20%, 2.14%, and 3.85% and 4.30%, 4.00%, and 4.40%, respectively. The limit of detection was 0.10 kUA/L, and the limit of quantitation was 0.11 kUA/L. The range of linearity was from 0.27 kUA/L to 97.53 kUA/L (r = 0.9968). The correlation coefficient (r) for the correlation analysis between results of LICA and capture ELISA was 0.9087. This assay was successfully applied in 64 human serum samples, showing good sensitivity (82.20%) and specificity (100%). CONCLUSION: An Artemisia-sIgE quantitation assay based on LICA was successfully established. Its performance satisfied the clinical requirements and could be widely used in clinical laboratories.


Assuntos
Artemisia , Alérgenos , Imunoglobulina E , Luminescência , Medições Luminescentes/métodos
16.
Anal Bioanal Chem ; 414(28): 8081-8091, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-36152037

RESUMO

A simple, sensitive, specific and fast method based on the loop-mediated isothermal amplification (LAMP) technique and cleavable molecular beacon (CMB) was developed for chicken authentication detection. LAMP and CMB were used for DNA amplification and amplicon analysis, respectively. Targeting the mitochondrial cytochrome b gene of chickens, five primers and one CMB probe were designed, and their specificity was validated against nine other animal species. The structure of CMB and concentrations of dNTPs, MgSO4, betaine, RNase H2, primers and CMB were optimized. The CMB-LAMP assay was completed within 17 min, and its limit of detection for chicken DNA was 1.5 pg µL-1. Chicken adulteration as low as 0.5% was detected in beef, and no cross-reactivity was observed. Finally, this assay was successfully applied to 20 commercial meat products. When combined with our developed DNA extraction method (the extraction time was 1 min: lysis for 10 s, washing for 20 s and elution for 30 s), the entire process (from DNA extraction to results analysis) was able to be completed within 20 min, which is at least 10 min shorter than other LAMP-based methods. Our method showed great potential for the on-site detection of chicken adulteration in meat.


Assuntos
Galinhas , Técnicas de Amplificação de Ácido Nucleico , Bovinos , Animais , Técnicas de Amplificação de Ácido Nucleico/métodos , Carne/análise , Primers do DNA/genética , DNA , Sensibilidade e Especificidade
17.
Indian J Med Res ; 155(5&6): 565-569, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36124504

RESUMO

Background & objectives: The pandemic caused by the SARS-CoV-2 has been a threat to humankind due to the rapid spread of infection and appearance of multiple new variants. In the present study, we report the dynamics and persistence of immunoglobulin G (IgG) and immunoglobulin M (IgM) antibodies in asymptomatic and symptomatic COVID-19 patients by chemiluminescent assay. Methods: A total of 463 serum samples from 218 SARS-CoV-2 PCR-positive patients were collected over a period of 124 days post-onset of disease (POD). Antibody levels were measured by chemiluminescence bioanalyzer. Neutralizing antibody titres were assessed by plaque reduction neutralization test (PRNT) for SARS-CoV-2. Results: Both IgM and IgG started appearing from day five post-infection in symptomatic and asymptomatic patients. IgM antibody response peaked around day 35 POD and rapidly diminished thereafter, with the last IgM-positive sample observed at 90 days POD. IgG antibody response peaked around 45 days POD and persisted till 124 days. The chemiluminescence immunoassay (CLIA) results showed a moderate correlation (R=0.5846, P<0.001) compared with PRNT. Additional analysis indicated a neutralizing titre of 250 corresponded to 12.948 AU/ml of YHLO iFlash SARS-CoV-2 IgG units. Interpretation & conclusions: Both symptomatic and asymptomatic COVID-19 patients seem to initiate production of antibody responses from day five of onset of disease. Although the CLIA gives high sensitivity and specificity and also its binding IgG antibody titres may correlate moderately with protective immunity, our results indicate that the values of binding antibody alone may not be a perfect guide to represent virus neutralization titre during donor selection for plasma therapy. However, IgM and IgG antibody detection may help in monitoring the status of disease progression and burden in the community.


Assuntos
COVID-19 , Humanos , SARS-CoV-2 , Anticorpos Antivirais , Imunoglobulina M , Imunoglobulina G , Sensibilidade e Especificidade
18.
J Clin Lab Anal ; 36(1): e24069, 2022 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-34783398

RESUMO

BACKGROUND: This study aimed to investigate the implementation and quality control of the quantitative detection of serum Helicobacter pylori (H. pylori) antibody in clinical laboratories in China. METHODS: Online external quality assessment (EQA) questionnaires were distributed to the clinical laboratories by National Center for Clinical Laboratories (NCCL) of China. We collected information on the quantitative detection procedures of serum H. pylori antibody in clinical laboratories, including detection reagents, methods, instruments, calibrators, and internal quality control (IQC). We distributed quality control products to some select laboratories that conducted quantitative detection and analyzed the obtained test data. We evaluated the quantitative detection procedure based on the standard evaluation criteria set at a target value of ±30%. RESULTS: 70.9% (146/206) of the laboratories conducted quantitative detection of H. pylori antibody; 29.1% (60/206) of the laboratories performed qualitative detection. Domestic reagents and matching calibrators accounted for more than 97.1% (200/206) of all reagents. Latex-enhanced immunoturbidimetry was used in 89.7% (131/146) of the laboratories for quantitative determination, while the colloidal gold method was used in 66.7% (40/60) of the laboratories for qualitative determination. A total of 130 laboratories participated in the EQA; 123 completed the assessment, and the pass rate was 75.6% (93/123). CONCLUSION: Clinical quantitative detection of serum H. pylori antibody is performed at a high rate in China. Thus, further studies on the specificity of commercial detection reagents are needed. EQAs are useful to monitor and improve the detection quality of H. pylori antibodies.


Assuntos
Anticorpos Antibacterianos/sangue , Helicobacter pylori/imunologia , Laboratórios Clínicos , China , Infecções por Helicobacter/diagnóstico , Infecções por Helicobacter/imunologia , Infecções por Helicobacter/microbiologia , Humanos , Imunoturbidimetria/normas , Laboratórios Clínicos/normas , Laboratórios Clínicos/estatística & dados numéricos , Controle de Qualidade , Inquéritos e Questionários
19.
J Clin Lab Anal ; 36(7): e24490, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-35587485

RESUMO

AIM: To explore the quality control and implementation of the quantitative detection of liver fibrosis biomarkers, laminin (LN), collagen IV (Col Ⅳ), procollagen III amino-terminal propeptide (PⅢNP), hyaluronic acid (HA), and cholyglycine (CG), in China. METHODS: Two quality control products were measured in different laboratories using different measurement methods and reagents, and the acquired results were subjected to analysis. The quantitative detection technique was based on the conventional assessment criteria, with a target value ±30% being employed. RESULTS: Hundred labs were involved in the External Quality Assessment with 88 laboratories completing the assessment, and the pass rates were 84%, 80.2%, 67.5%, 77.3%, and 58.3% for HA, LN, PⅢNP, Col Ⅳ, and CG, respectively. Chemiluminescence immunoassay was used most for HA (90.1%), LN (90.1%), PⅢNP (87.9%), and Col Ⅳ (82.9%) determination, whereas the chemiluminescence immunoassay (31.6%), latex-enhanced immunoturbidimetry (36.7%), and homogeneous enzyme immunoassay (26.7%) were used for CG determination. The coefficients of variation for HA, LN, PⅢNP, Col Ⅳ, and CG in different laboratories were 3.3%-19.49%, 1.74%-38.81%, 1.97%-41.29%, 2.85%-41.69%, and 2.71%-41.8%, respectively. CONCLUSION: The clinical quantitative detection of liver fibrosis biomarkers is highly performed in China. The existing problems are that there are many manufacturers producing reagents and instruments, the quality of reagents is uneven, the specificity and sensitivity of reagents are greatly different, the comparability of results of various systems is poor, and the accuracy and consistency between different systems are lacking. All above underscores the critical importance of EQA in improving and monitoring the identification of biomarkers for liver fibrosis.


Assuntos
Laminina , Cirrose Hepática , Biomarcadores , Colágeno Tipo IV , Humanos , Ácido Hialurônico , Cirrose Hepática/diagnóstico
20.
Mikrochim Acta ; 189(6): 218, 2022 05 11.
Artigo em Inglês | MEDLINE | ID: mdl-35546375

RESUMO

A dual-signal photometric/fluorometric assay was established for rapid, qualitative, and quantitative detection of Salmonella typhimurium (S. typhimurium). This method was composed of two parts: (1) a single-step photometric (SSC) assay containing gold nanoparticles (AuNPs), poly-diallyldimethylammonium chloride (PDDA), and S. typhimurium-specific aptamer, and (2) a fluorescence (FL) assay containing carboxyl-modified CdSe/ZnS quantum dots (QDs-COOH). Users just need to drop samples contaminated with S. typhimurium into SSC assay; the apparent color change from red to blue can be observed in a short time (20 min). A smartphone app was developed to read the semiquantitative result. By subsequently adding one drop of FL assay into the reaction mixture, the generated fluorescence intensity reflected the concentration of S. typhimurium. The naked eye limit of detection (LOD) and fluorescent LOD were 103 cfu/mL and 10 cfu/mL, respectively. This method exhibited good selectivity. The reliability and practicability were verified by testing contaminated food, drinking water, and pets' urine.


Assuntos
Ouro , Nanopartículas Metálicas , Limite de Detecção , Reprodutibilidade dos Testes , Salmonella typhimurium
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