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1.
Int J Mol Sci ; 24(11)2023 May 26.
Article in English | MEDLINE | ID: covidwho-20232955

ABSTRACT

The term "liver disease" refers to any hepatic condition that leads to tissue damage or altered hepatic function and can be induced by virus infections, autoimmunity, inherited genetic mutations, high consumption of alcohol or drugs, fat accumulation, and cancer. Some types of liver diseases are becoming more frequent worldwide. This can be related to increasing rates of obesity in developed countries, diet changes, higher alcohol intake, and even the coronavirus disease 2019 (COVID-19) pandemic was associated with increased liver disease-related deaths. Although the liver can regenerate, in cases of chronic damage or extensive fibrosis, the recovery of tissue mass is impossible, and a liver transplant is indicated. Because of reduced organ availability, it is necessary to search for alternative bioengineered solutions aiming for a cure or increased life expectancy while a transplant is not possible. Therefore, several groups were studying the possibility of stem cells transplantation as a therapeutic alternative since it is a promising strategy in regenerative medicine for treating various diseases. At the same time, nanotechnological advances can contribute to specifically targeting transplanted cells to injured sites using magnetic nanoparticles. In this review, we summarize multiple magnetic nanostructure-based strategies that are promising for treating liver diseases.


Subject(s)
COVID-19 , Liver Diseases , Nanostructures , Humans , Regenerative Medicine , Hepatocytes/transplantation , COVID-19/therapy , Liver Diseases/therapy , Stem Cells , Liver Regeneration , Magnetic Phenomena
2.
Biosensors (Basel) ; 13(5)2023 May 18.
Article in English | MEDLINE | ID: covidwho-20242365

ABSTRACT

COVID-19 has resulted in a pandemic that aggravated the world's healthcare systems, economies, and education, and caused millions of global deaths. Until now, there has been no specific, reliable, and effective treatment to combat the virus and its variants. The current standard tedious PCR-based tests have limitations in terms of sensitivity, specificity, turnaround time, and false negative results. Thus, an alternative, rapid, accurate, and sensitive diagnostic tool that can detect viral particles, without the need for amplification or viral replication, is central to infectious disease surveillance. Here, we report MICaFVi (Magnetic Immuno-Capture Flow Virometry), a novel precise nano-biosensor diagnostic assay for coronavirus detection which combines the MNP-based immuno-capture of viruses for enrichment followed by flow-virometry analysis, enabling the sensitive detection of viral particles and pseudoviruses. As proof of concept, virus-mimicking spike-protein-coated silica particles (VM-SPs) were captured using anti-spike-antibody-conjugated MNPs (AS-MNPs) followed by detection using flow cytometry. Our results showed that MICaFVi can successfully detect viral MERS-CoV/SARS-CoV-2-mimicking particles as well as MERS-CoV pseudoviral particles (MERSpp) with high specificity and sensitivity, where a limit of detection (LOD) of 3.9 µg/mL (20 pmol/mL) was achieved. The proposed method has great potential for designing practical, specific, and point-of-care testing for rapid and sensitive diagnoses of coronavirus and other infectious diseases.


Subject(s)
COVID-19 , Middle East Respiratory Syndrome Coronavirus , Humans , COVID-19/diagnosis , SARS-CoV-2 , COVID-19 Testing , Magnetic Phenomena
3.
Anal Chim Acta ; 1271: 341469, 2023 Aug 29.
Article in English | MEDLINE | ID: covidwho-20230823

ABSTRACT

Traditional nucleic acid extraction and detection is based on open operation, which may cause cross-contamination and aerosol formation. This study developed a droplet magnetic-controlled microfluidic chip integrated nucleic acid extraction, purification and amplification. The reagent is sealed in oil to form a droplet, and the nucleic acid is extracted and purified by controlling the movement of the magnetic beads (MBs) through a permanent magnet, ensuring a closed environment. This chip can automatically extract nucleic acid from multiple samples within 20 min, and can be directly placed in the in situ amplification instrument for amplification without further transfer of nucleic acid, characterized by simple, fast, time-saving and labor-saving. The results showed that the chip was able to detect <10 copies/test SARS-CoV-2 RNA, and EGFR exon 21 L858R mutations were detected in H1975 cells as low as 4 cells. In addition, on the basis of the droplet magnetic-controlled microfluidic chip, we further developed a multi-target detection chip, which used MBs to divide the nucleic acid of the sample into three parts. And the macrolides resistance mutations A2063G and A2064G, and the P1 gene of mycoplasma pneumoniae (MP) were successfully detected in clinical samples by the multi-target detection chip, providing the possibility for future application in the detection of multiple pathogens.


Subject(s)
COVID-19 , Neoplasms , Nucleic Acids , Humans , Nucleic Acids/genetics , Microfluidics , RNA, Viral , Nucleic Acid Amplification Techniques/methods , COVID-19/diagnosis , SARS-CoV-2 , Magnetic Phenomena
4.
Sensors (Basel) ; 23(9)2023 Apr 30.
Article in English | MEDLINE | ID: covidwho-2318020

ABSTRACT

Since its first report in 2006, magnetic particle spectroscopy (MPS)-based biosensors have flourished over the past decade. Currently, MPS are used for a wide range of applications, such as disease diagnosis, foodborne pathogen detection, etc. In this work, different MPS platforms, such as dual-frequency and mono-frequency driving field designs, were reviewed. MPS combined with multi-functional magnetic nanoparticles (MNPs) have been extensively reported as a versatile platform for the detection of a long list of biomarkers. The surface-functionalized MNPs serve as nanoprobes that specifically bind and label target analytes from liquid samples. Herein, an analysis of the theories and mechanisms that underlie different MPS platforms, which enable the implementation of bioassays based on either volume or surface, was carried out. Furthermore, this review draws attention to some significant MPS platform applications in the biomedical and biological fields. In recent years, different kinds of MPS point-of-care (POC) devices have been reported independently by several groups in the world. Due to the high detection sensitivity, simple assay procedures and low cost per run, the MPS POC devices are expected to become more widespread in the future. In addition, the growth of telemedicine and remote monitoring has created a greater demand for POC devices, as patients are able to receive health assessments and obtain results from the comfort of their own homes. At the end of this review, we comment on the opportunities and challenges for POC devices as well as MPS devices regarding the intensely growing demand for rapid, affordable, high-sensitivity and user-friendly devices.


Subject(s)
Biosensing Techniques , Point-of-Care Systems , Humans , Biosensing Techniques/methods , Magnetics , Spectrum Analysis , Magnetic Phenomena
5.
Appl Microbiol Biotechnol ; 107(10): 3329-3339, 2023 May.
Article in English | MEDLINE | ID: covidwho-2295184

ABSTRACT

Pandemics like SARS-Cov-2 very frequently have their origin in different animals and in particular herds of camels could be a source of zoonotic diseases. This study took advantage on a highly sensitive and adaptable method for the fast and reliable detection of viral antibodies in camels using low-cost equipment. Magnetic nanoparticles (MNP) have high variability in their functionalization with different peptides and proteins. We confirm that 3-aminopropyl triethoxysilane (APTES)-coated MNP could be functionalized with viral proteins. The protein loading could be confirmed by simple loading controls using FACS-analysis (p < 0.05). Complementary combination of antigen and antibody yields in a significant signal increase could be proven by both FACS and COMPASS. However, COMPASS needs only a few seconds for the measurement. In COMPASS, the phase φn on selected critical point of the fifth higher harmonic (n = 5th). Here, positive sera display highly significant signal increase over the control or negative sera. Furthermore, a clear distinction could be made in antibody detection as an immune response to closely related viruses (SARS-CoV2 and MERS). Using modified MNPs along with COMPASS offers a fast and reliable method that is less cost intensive than current technologies and offers the possibility to be quickly adapted in case of new occurring viral infections. KEY POINTS: • COMPASS (critical offset magnetic particle spectroscopy) allows the fast detection of antibodies. • Magnetic nanoparticles can be adapted by exchange of the linked bait molecule. • Antibodies could be detected in camel sera without washing steps within seconds.


Subject(s)
COVID-19 , Middle East Respiratory Syndrome Coronavirus , Animals , Antibodies, Viral , Camelus , RNA, Viral , Middle East Respiratory Syndrome Coronavirus/genetics , SARS-CoV-2 , Spectrum Analysis , Magnetic Phenomena
6.
Biosensors (Basel) ; 12(1)2021 Dec 23.
Article in English | MEDLINE | ID: covidwho-2250597

ABSTRACT

Sensitive serological assays are needed to provide valuable information about acute and past viral infections. For example, detection of anti-severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) IgG antibodies could serve as the basis for an "immunity passport" that would enable individuals to travel internationally. Here, utilizing a novel Magnetic Modulation Biosensing (MMB) system and the receptor-binding domain of the SARS-CoV-2 spike protein, we demonstrate a highly sensitive and specific anti-SARS-CoV-2 IgG serological assay. Using anti-SARS-CoV-2 IgG antibodies, RT-qPCR SARS-CoV-2-positive and healthy patients' samples, and vaccinees' samples, we compare the MMB-based SARS-CoV-2 IgG assay's analytical and clinical sensitivities to those of the enzyme-linked immunosorbent assay (ELISA). Compared with ELISA, the MMB-based assay has an ~6-fold lower limit of detection (129 ng/L vs. 817 ng/L), and it detects an increase in the IgG concentration much earlier after vaccination. Using 85 RT-qPCR SARS-CoV-2-positive samples and 79 -negative samples, the MMB-based assay demonstrated similar clinical specificity (98% vs. 99%) and sensitivity (93% vs. 92%) to the ELISA test, but with a much faster turnaround time (45 min vs. 245 min). The high analytical and clinical sensitivity, short turnaround time, and simplicity of the MMB-based assay makes it a preferred method for antibody detection.


Subject(s)
Antibodies, Viral/analysis , Biosensing Techniques , COVID-19 , Immunoglobulin G/analysis , Serologic Tests , COVID-19/diagnosis , COVID-19/immunology , Enzyme-Linked Immunosorbent Assay , Humans , Magnetic Phenomena , SARS-CoV-2/immunology , Sensitivity and Specificity , Spike Glycoprotein, Coronavirus
7.
Anal Chem ; 95(14): 6182-6190, 2023 04 11.
Article in English | MEDLINE | ID: covidwho-2289077

ABSTRACT

Pooling multiple samples prior to real-time reverse-transcription polymerase chain reaction (RT-PCR) analysis has been proposed as a strategy to minimize expenses and boost test throughput during the COVID-19 pandemic. Nevertheless, the traditional pooling approach cannot be effectively deployed in high-prevalence settings due to the need for secondary tests in the case of a positive pool. In this study, we present a pooling test platform with high adaptability and simplicity that allows sample-specific detection of multiple-tagged samples in a single run without the need for retesting. This was accomplished by labeling distinct samples with predefined ID-Primers and identifying tagged pooled samples using one-step RT-PCR followed by melting curve analysis with rationally designed universal fluorescence- and quencher-tagged oligo probes. Using magnetic beads (MBs), nucleic acid targets from different individuals can be tagged and extracted concurrently and then pooled before RT, eliminating the need for extra RNA extraction and separate RT and enzyme digestion steps in the recently developed barcoding strategies. Pools of six samples (positive and negative) were successfully identified by melting temperature values under two fluorescent channels, with a detection sensitivity of 5 copies/µL. We validated the reproducibility of this assay by running it on 40 clinical samples with a hypothetical infection rate of 15%. In addition, to aid the scenario of large-scale pooling tests, we constructed a melting curve autoreadout system (MCARS) for statistical analysis of melting curve plots to eliminate error-prone manual result readout. Our results suggest that this strategy could be a simple and adaptable tool for alleviating existing bottlenecks in diagnostic pooling testing.


Subject(s)
COVID-19 , Humans , Pandemics , Reproducibility of Results , COVID-19 Testing , Magnetic Phenomena , Sensitivity and Specificity , RNA, Viral/genetics
8.
Nanotechnology ; 34(27)2023 Apr 19.
Article in English | MEDLINE | ID: covidwho-2260470

ABSTRACT

Infectious diseases such as novel coronavirus (SARS-CoV-2), Influenza, HIV, Ebola, etc kill many people around the world every year (SARS-CoV-2 in 2019, Ebola in 2013, HIV in 1980, Influenza in 1918). For example, SARS-CoV-2 has plagued higher than 317 000 000 people around the world from December 2019 to January 13, 2022. Some infectious diseases do not yet have not a proper vaccine, drug, therapeutic, and/or detection method, which makes rapid identification and definitive treatments the main challenges. Different device techniques have been used to detect infectious diseases. However, in recent years, magnetic materials have emerged as active sensors/biosensors for detecting viral, bacterial, and plasmids agents. In this review, the recent applications of magnetic materials in biosensors for infectious viruses detection have been discussed. Also, this work addresses the future trends and perspectives of magnetic biosensors.


Subject(s)
Biosensing Techniques , COVID-19 , Communicable Diseases , Ebolavirus , HIV Infections , Hemorrhagic Fever, Ebola , Influenza, Human , Humans , SARS-CoV-2 , COVID-19/diagnosis , Magnetic Phenomena
9.
Nano Lett ; 23(7): 2636-2643, 2023 04 12.
Article in English | MEDLINE | ID: covidwho-2254626

ABSTRACT

Biomolecular interactions compose a fundamental element of all life forms and are the biological basis of many biomedical assays. However, current methods for detecting biomolecular interactions have limitations in sensitivity and specificity. Here, using nitrogen-vacancy centers in diamond as quantum sensors, we demonstrate digital magnetic detection of biomolecular interactions with single magnetic nanoparticles (MNPs). We first developed a single-particle magnetic imaging (SiPMI) method on 100 nm-sized MNPs with negligible magnetic background, high signal stability, and accurate quantification. The single-particle method was performed on biotin-streptavidin interactions and DNA-DNA interactions in which a single-base mismatch was specifically differentiated. Subsequently, SARS-CoV-2-related antibodies and nucleic acids were examined by a digital immunomagnetic assay derived from SiPMI. In addition, a magnetic separation process improved the detection sensitivity and dynamic range by more than 3 orders of magnitude and also the specificity. This digital magnetic platform is applicable to extensive biomolecular interaction studies and ultrasensitive biomedical assays.


Subject(s)
COVID-19 , Nanoparticles , Humans , SARS-CoV-2 , DNA , Magnetic Phenomena
10.
J Med Internet Res ; 25: e43497, 2023 03 31.
Article in English | MEDLINE | ID: covidwho-2248351

ABSTRACT

BACKGROUND: The popularity of the magnetic vaccine conspiracy theory and other conspiracy theories of a similar nature creates challenges to promoting vaccines and disseminating accurate health information. OBJECTIVE: Health conspiracy theories are gaining in popularity. This study's objective was to evaluate the Twitter social media network related to the magnetic vaccine conspiracy theory and apply social capital theory to analyze the unique social structures of influential users. As a strategy for web-based public health surveillance, we conducted a social network analysis to identify the important opinion leaders sharing the conspiracy, the key websites, and the narratives. METHODS: A total of 18,706 tweets were retrieved and analyzed by using social network analysis. Data were retrieved from June 1 to June 13, 2021, using the keyword vaccine magnetic. Tweets were retrieved via a dedicated Twitter application programming interface. More specifically, the Academic Track application programming interface was used, and the data were analyzed by using NodeXL Pro (Social Media Research Foundation) and Gephi. RESULTS: There were a total of 22,762 connections between Twitter users within the data set. This study found that the most influential user within the network consisted of a news account that was reporting on the magnetic vaccine conspiracy. There were also several other users that became influential, such as an epidemiologist, a health economist, and a retired sports athlete who exerted their social capital within the network. CONCLUSIONS: Our study found that influential users were effective broadcasters against the conspiracy, and their reach extended beyond their own networks of Twitter followers. We emphasize the need for trust in influential users with regard to health information, particularly in the context of the widespread social uncertainty resulting from the COVID-19 pandemic, when public sentiment on social media may be unpredictable. This study highlights the potential of influential users to disrupt information flows of conspiracy theories via their unique social capital.


Subject(s)
COVID-19 , Social Media , Vaccines , Humans , Pandemics , Social Network Analysis , Magnetic Phenomena
11.
Sci Rep ; 13(1): 2163, 2023 02 07.
Article in English | MEDLINE | ID: covidwho-2227499

ABSTRACT

Presented here is a magnetic hydrogel particle enabled workflow for capturing and concentrating SARS-CoV-2 from diagnostic remnant swab samples that significantly improves sequencing results using the Oxford Nanopore Technologies MinION sequencing platform. Our approach utilizes a novel affinity-based magnetic hydrogel particle, circumventing low input sample volumes and allowing for both rapid manual and automated high throughput workflows that are compatible with Nanopore sequencing. This approach enhances standard RNA extraction protocols, providing up to 40 × improvements in viral mapped reads, and improves sequencing coverage by 20-80% from lower titer diagnostic remnant samples. Furthermore, we demonstrate that this approach works for contrived influenza virus and respiratory syncytial virus samples, suggesting that it can be used to identify and improve sequencing results of multiple viruses in VTM samples. These methods can be performed manually or on a KingFisher automation platform.


Subject(s)
COVID-19 , Nanopore Sequencing , Humans , SARS-CoV-2 , Nanopore Sequencing/methods , Hydrogels , High-Throughput Nucleotide Sequencing/methods , Magnetic Phenomena
12.
ACS Appl Mater Interfaces ; 15(4): 4924-4934, 2023 Feb 01.
Article in English | MEDLINE | ID: covidwho-2229927

ABSTRACT

Although numerous approaches were proposed for the nucleic acid (NA)-based SARS-CoV-2 detection, the nonideal NA desorption efficiency of conventional magnetic beads (MBs) limits their widespread application. In this study, we developed solvent-responsive MBs (called responsive MBs), which, in the presence of buffers, modulated the absorption and desorption capacities of NA by flipping the surface -COO-. Relative to other commercial MBs, responsive MBs exhibited similar absorption profiles and markedly enhanced desorption profiles. When applied for NA detection of complex samples, responsive MBs exhibited better performance of RNA detection than DNA, with obvious advantages in sensitivity. Specifically, the RNA and DNA desorption rates of commercial MBs were ∼85 and 82.5%, while those of responsive MBs were nearly 94 and 93.5%, respectively. Furthermore, responsive MBs exhibited remarkable extraction ability in a wide range of tissues and better performance of RNA extraction than DNA. When applied for SARS-CoV-2 detection, the responsive MBs along with the simulated digital RT-LAMP (a previously established apparatus) further improved detection efficiency, yielding a precise quantitative detection as low as 25 copies and an ultimate sensibility detection of 5 copies/mL. It was also successfully employed in numerous NA-based technologies such as polymerase chain reaction (PCR), sequencing, and so on.


Subject(s)
COVID-19 , SARS-CoV-2 , Humans , COVID-19/diagnosis , COVID-19 Testing , Clinical Laboratory Techniques , Sensitivity and Specificity , Nucleic Acid Amplification Techniques , RNA, Viral/genetics , RNA, Viral/analysis , Real-Time Polymerase Chain Reaction , Magnetic Phenomena , DNA
13.
Talanta ; 256: 124271, 2023 May 01.
Article in English | MEDLINE | ID: covidwho-2183603

ABSTRACT

Rapid screening of multiple pathogens will greatly improve the efficiency of pandemic prevention and control. Colorimetric methods exhibit the advantages of convenience, portability, low cost, time efficiency, and free of sophisticated instruments, yet usually have difficulties in simultaneous detection and suffer from monotonous color changes with low visual resolution and sensitivity. Hence, coupled three kinds of plasmonic nanoparticles (NPs) with magnetic separation, we developed an achromatic colorimetric nanosensor with highly enhanced visual resolution for simultaneous detection of SARS-CoV-2, Staphylococcus aureus, and Salmonella typhimurium. The achromatic nanosensor was composed of SARS-CoV-2-targeting red gold NPs, S. aureus-targeting yellow silver NPs and S. typhimurium-targeting blue silver triangle NPs mixed as black color. In the detection, three corresponding magnetic probes were added into the above mixture. In the presence of a target pathogen, it would be recognized and combined with corresponding colored reporters and magnetic probes to form sandwich complexes, which were removed by magnetic separation, and the sensor changed from black to a chromatic color (the color of the reporters remained in supernatant). Consequently, different target pathogen induced different color. For example, SARS-CoV-2, S. aureus, and S. typhimurium respectively produced green, purple, and orange. While coexistence of S. aureus and S. typhimurium produced red, and coexistence of S. aureus and SARS-CoV-2 produced blue, etc. Therefore, by observing the color change or measuring the absorption spectra, multiple pathogen detection was achieved conveniently. Compared with most colorimetric sensors, this achromatic nanosensor involved rich color change, thus significantly enhancing visual resolution and inspection sensitivity. Therefore, this sensor opened a promising avenue for efficient monitoring and early warning of food safety and quality.


Subject(s)
COVID-19 , Metal Nanoparticles , Nanoparticles , Humans , Silver , Colorimetry/methods , Staphylococcus aureus , COVID-19/diagnosis , SARS-CoV-2 , Gold , Magnetic Phenomena
14.
Bioelectrochemistry ; 150: 108358, 2023 Apr.
Article in English | MEDLINE | ID: covidwho-2165106

ABSTRACT

A sensitive, reliable, and cost-effective detection for SARS-CoV-2 was urgently needed due to the rapid spread of COVID-19. Here, a "signal-on" magnetic-assisted PEC immunosensor was constructed for the quantitative detection of SARS-CoV-2 nucleocapsid (N) protein based on Z-scheme heterojunction. Fe3O4@SiO2@Au was used to connect the capture antibody to act as a capture probe (Fe3O4@SiO2@Au/Ab1). It can extract target analytes selectively in complex samples and multiple electrode rinsing and assembly steps were avoided effectively. CdTe QDs sensitized TiO2 coated on the surface of SiO2 spheres to form Z-scheme heterojunction (SiO2@TiO2@CdTe QDs), which broadened the optical absorption range and inhibited the quick recombination of photogenerated electron/hole of the composite. With fascinating photoelectric conversion performance, SiO2@TiO2@CdTe QDs were utilized as a signal label, thus further realizing signal amplification. The migration mechanism of photogenerated electrons was further deduced by active material quenching experiment and electron spin resonance (ESR) measurement. The elaborated immunosensor can detect SARS-CoV-2 N protein in the linear range of 0.005-50 ng mL-1 with a low detection limit of 1.8 pg mL-1 (S/N = 3). The immunosensor displays extraordinary sensitivity, strong anti-interference, and high reproducibility in detecting SARS-CoV-2 N protein, which envisages its potential application in the clinical diagnosis of COVID-19.


Subject(s)
Biosensing Techniques , COVID-19 , Cadmium Compounds , Nanocomposites , Quantum Dots , Humans , COVID-19/diagnosis , Electrochemical Techniques , Immunoassay , Limit of Detection , Magnetic Phenomena , Nucleocapsid Proteins , Reproducibility of Results , SARS-CoV-2 , Silicon Dioxide , Tellurium
15.
Nat Commun ; 13(1): 7230, 2022 Nov 24.
Article in English | MEDLINE | ID: covidwho-2133434

ABSTRACT

Magnetic nanoparticles (MNPs) have been adapted for many applications, e.g., bioassays for the detection of biomarkers such as antibodies, by controlled engineering of specific surface properties. Specific measurement of such binding states is of high interest but currently limited to highly sensitive techniques such as ELISA or flow cytometry, which are relatively inflexible, difficult to handle, expensive and time-consuming. Here we report a method named COMPASS (Critical-Offset-Magnetic-Particle-SpectroScopy), which is based on a critical offset magnetic field, enabling sensitive detection to minimal changes in mobility of MNP ensembles, e.g., resulting from SARS-CoV-2 antibodies binding to the S antigen on the surface of functionalized MNPs. With a sensitivity of 0.33 fmole/50 µl (≙7 pM) for SARS-CoV-2-S1 antibodies, measured with a low-cost portable COMPASS device, the proposed technique is competitive with respect to sensitivity while providing flexibility, robustness, and a measurement time of seconds per sample. In addition, initial results with blood serum demonstrate high specificity.


Subject(s)
COVID-19 , Magnetite Nanoparticles , Humans , Magnetite Nanoparticles/chemistry , COVID-19/diagnosis , SARS-CoV-2 , Spectrum Analysis , Antibodies, Viral , Point-of-Care Testing , Magnetic Phenomena
16.
J Vet Med Sci ; 84(11): 1543-1550, 2022 Nov 14.
Article in English | MEDLINE | ID: covidwho-2065087

ABSTRACT

In this study, the viral genome extraction performance of automatic nucleic acid extractors and manual nucleic acid extraction kits was compared. We showed that compared with manual kits, the automatic extractors showed superior genome extraction performance using bovine viral diarrhea virus (BVDV) genome-positive cattle sera and bovine coronavirus/infectious bovine rhinotracheitis virus-spiked cattle nasal swabs. In addition, the subgenotyping of BVDV strains detected in Tokachi Province in Japan during 2016-2017 was performed. Results showed that most of these BVDV strains belonged to subgenotype 1b, while few strains belonged to subgenotypes 1a and 2a. This study showed the high applicability of automatic nucleic acid extractors in extracting multiple viral genomes and the dominant subgenotype of BVDV in Tokachi.


Subject(s)
Bovine Virus Diarrhea-Mucosal Disease , Cattle Diseases , Diarrhea Virus 1, Bovine Viral , Diarrhea Viruses, Bovine Viral , Nucleic Acids , Cattle , Animals , RNA, Viral/genetics , Japan , Genotype , Diarrhea Viruses, Bovine Viral/genetics , Diarrhea/veterinary , Magnetic Phenomena , Diarrhea Virus 1, Bovine Viral/genetics , Phylogeny
17.
Anal Chim Acta ; 1233: 340486, 2022 Nov 15.
Article in English | MEDLINE | ID: covidwho-2060277

ABSTRACT

The SARS-CoV-2 pandemic has posed a huge challenge to rapid and accurate diagnosis of SARS-CoV-2 in the early stage of infection. In this work, we developed a novel magnetic/fluorescent dual-modal lateral flow immunoassay (LFIA) based on multifunctional nanobeads for rapid and accurate determination of SARS-CoV-2 nucleocapsid protein (NP). The multifunctional nanobeads were fabricated by using polyethyleneimine (PEI) as a mediate shell to combine superparamagnetic Fe3O4 core with dual quantum dot shells (MagDQD). The core-shell structure of MagDQD label with high loading density of quantum dots (QDs) and superior magnetic content realized LFIA with dual quantitative analysis modal from the assemblies of individual single nanoparticles. The LFIA integrated the advantages of magnetic signal and fluorescent signal, resulting excellent accuracy for quantitative analysis and high elasticity of the overall detection. In addition, magnetic signal and fluorescent signal both had high sensitivity with the limit of detection (LOD) as 0.235 ng mL-1 and 0.012 ng mL-1, respectively. The recovery rates of the methods in simulated saliva samples were 91.36%-103.60% (magnetic signal) and 94.39%-104.38% (fluorescent signal). The results indicate the method has a considerable potential to be an effective tool for diagnose SARS-CoV-2 in the early stage of infection.


Subject(s)
COVID-19 , SARS-CoV-2 , Humans , Polyethyleneimine , COVID-19/diagnosis , Immunoassay/methods , Magnetic Phenomena
18.
Braz J Microbiol ; 53(3): 1263-1269, 2022 Sep.
Article in English | MEDLINE | ID: covidwho-2014657

ABSTRACT

Immunological assays to detect SARS-CoV-2 Spike Receptor Binding Domain (RBD) antigen seroconversion in humans are important tools to monitor the levels of protecting antibodies in the population in response to infection and/or immunization. Here we describe a simple, low cost, and high throughput Ni2+ magnetic bead immunoassay to detect human IgG reactive to Spike S1 RBD Receptor Binding Domain produced in Escherichia coli. A 6xHis-tagged Spike S1 RBD was expressed in E. coli and purified by affinity chromatography. The protein was mobilized on the surface of Ni2+ magnetic beads and used to investigate the presence of reactive IgG in the serum obtained from pre-pandemic and COVID-19 confirmed cases. The method was validated with a cohort of 290 samples and an area under the receiver operating characteristic curve of 0.94 was obtained. The method was operated with > 82% sensitivity at 98% specificity and was also able to track human IgG raised in response to vaccination with Comirnaty at > 85% sensitivity. The IgG signal obtained with the described method was well-correlated with the signal obtained when pre fusion Spike produced in HEK cell lines was used as antigen. This novel low-cost and high throughput immunoassay may act as an important tool to investigate protecting IgG antibodies against SARS-CoV-2 in the human population.


Subject(s)
COVID-19 , SARS-CoV-2 , Antibodies, Viral , Escherichia coli/genetics , Humans , Immunoassay/methods , Immunoglobulin G , Magnetic Phenomena , SARS-CoV-2/genetics , Spike Glycoprotein, Coronavirus/genetics
20.
Mikrochim Acta ; 189(4): 168, 2022 04 01.
Article in English | MEDLINE | ID: covidwho-1767504

ABSTRACT

The rapid spread of the novel human coronavirus 2019 (COVID-19) and its morbidity have created an urgent need for rapid and sensitive diagnostics. The real-time polymerase chain reaction is the gold standard for detecting the coronavirus in various types of biological specimens. However, this technique is time consuming, labor intensive, and expensive. Screen-printed electrodes (SPEs) can be used as point-of-care devices because of their low cost, sensitivity, selectivity, and ability to be miniaturized. The ability to detect the spike protein of COVID-19 in serum, urine, and saliva was developed using SPE aided by magnetic beads (MBs) and a portable potentiostat. The antibody-peroxidase-loaded MBs were the captured and catalytic units for the electrochemical assays. The MBs enable simple washing and homogenous deposition on the working electrode using a magnet. The assembly of the immunological MBs and the electrochemical system increases the measuring sensitivity and speed. The physical and electrochemical properties of the layer-by-layer modified MBs were systematically characterized. The performance of these immunosensors was evaluated using spike protein in the range 3.12-200 ng mL-1. We achieved a limit of detection of 0.20, 0.31, and 0.54 ng mL-1 in human saliva, urine, and serum, respectively. A facile electrochemical method to detect COVID-19 spike protein was developed for quick point-of-care testing.


Subject(s)
Biosensing Techniques , COVID-19 , Biosensing Techniques/methods , COVID-19/diagnosis , Electrodes , Humans , Immunoassay , Magnetic Phenomena , Point-of-Care Testing , Spike Glycoprotein, Coronavirus
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