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1.
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
2.
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
3.
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
4.
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
6.
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
7.
Biosensors (Basel) ; 12(3)2022 Mar 02.
Article in English | MEDLINE | ID: covidwho-1760369

ABSTRACT

Campylobacter jejuni is one of the most important causes of food-borne infectious disease, and poses challenges to food safety and public health. Establishing a rapid, accurate, sensitive, and simple detection method for C. jejuni enables early diagnosis, early intervention, and prevention of pathogen transmission. In this study, an immunocapture magnetic bead (ICB)-enhanced loop-mediated isothermal amplification (LAMP) CRISPR/Cas12a method (ICB-LAMP-CRISPR/Cas12a) was developed for the rapid and visual detection of C. jejuni. Using the ICB-LAMP-CRISPR/Cas12a method, C. jejuni was first captured by ICB, and the bacterial genomic DNA was then released by heating and used in the LAMP reaction. After the LAMP reaction, LAMP products were mixed and detected by the CRISPR/Cas12a cleavage mixture. This ICB-LAMP-CRISPR/Cas12a method could detect a minimum of 8 CFU/mL of C. jejuni within 70 min. Additionally, the method was performed in a closed tube in addition to ICB capture, which eliminates the need to separate preamplification and transfer of amplified products to avoid aerosol pollution. The ICB-LAMP-CRISPR/Cas12a method was further validated by testing 31 C. jejuni-positive fecal samples from different layer farms. This method is an all-in-one, simple, rapid, ultrasensitive, ultraspecific, visual detection method for instrument-free diagnosis of C. jejuni, and has wide application potential in future work.


Subject(s)
Campylobacter jejuni , CRISPR-Cas Systems , Magnetic Phenomena , Molecular Diagnostic Techniques , Nucleic Acid Amplification Techniques/methods
8.
Biomed Chromatogr ; 36(6): e5365, 2022 Jun.
Article in English | MEDLINE | ID: covidwho-1739127

ABSTRACT

Favipiravir is a potential antiviral medication that has been recently licensed for Covid-19 treatment. In this work, a gadolinium-based magnetic ionic liquid was prepared and used as an extractant in dispersive liquid-liquid microextraction (DLLME) of favipiravir in human plasma. The high enriching ability of DLLME allowed the determination of favipiravir in real samples using HPLC/UV with sufficient sensitivity. The effects of several variables on extraction efficiency were investigated, including type of extractant, amount of extractant, type of disperser and disperser volume. The maximum enrichment was attained using 50 mg of the Gd-magnetic ionic liquid (MIL) and 150 µl of tetrahydrofuran. The Gd-based MIL could form a supramolecular assembly in the presence of tetrahydrofuran, which enhanced the extraction efficiency of favipiravir. The developed method was validated according to US Food and Drug Administration bioanalytical method validation guidelines. The coefficient of determination was 0.9999, for a linear concentration range of 25 to 1.0 × 105  ng/ml. The percentage recovery (accuracy) varied from 99.83 to 104.2%, with RSD values (precision) ranging from 4.07 to 11.84%. The total extraction time was about 12 min and the HPLC analysis time was 5 min. The method was simple, selective and sensitive for the determination of favipiravir in real human plasma.


Subject(s)
COVID-19 , Ionic Liquids , Liquid Phase Microextraction , Amides , COVID-19/drug therapy , Chromatography, High Pressure Liquid/methods , Furans , Gadolinium , Humans , Liquid Phase Microextraction/methods , Magnetic Phenomena , Pyrazines
9.
J Mater Chem B ; 9(47): 9642-9657, 2021 12 08.
Article in English | MEDLINE | ID: covidwho-1684136

ABSTRACT

Cancer is a growing threat to human beings. Traditional treatments for malignant tumors usually involve invasive means to healthy human tissues, such as surgical treatment and chemotherapy. In recent years the use of specific stimulus-responsive materials in combination with some non-contact, non-invasive stimuli can lead to better efficacy and has become an important area of research. It promises to develop personalized treatment systems for four types of physical stimuli: light, ultrasound, magnetic field, and temperature. Nanomaterials that are responsive to these stimuli can be used to enhance drug delivery, cancer treatment, and tissue engineering. This paper reviews the principles of the stimuli mentioned above, their effects on materials, and how they work with nanomaterials. For this aim, we focus on specific applications in controlled drug release, cancer therapy, tissue engineering, and virus detection, with particular reference to recent photothermal, photodynamic, sonodynamic, magnetothermal, radiation, and other types of therapies. It is instructive for the future development of stimulus-responsive nanomaterials for these aspects.


Subject(s)
Antineoplastic Agents/therapeutic use , Delayed-Action Preparations/therapeutic use , Metal Nanoparticles/therapeutic use , Neoplasms/drug therapy , Radiation-Sensitizing Agents/therapeutic use , Animals , Antineoplastic Agents/chemistry , Antineoplastic Agents/radiation effects , Delayed-Action Preparations/chemistry , Delayed-Action Preparations/radiation effects , Humans , Infrared Rays , Magnetic Phenomena , Metal Nanoparticles/chemistry , Metal Nanoparticles/radiation effects , Radiation-Sensitizing Agents/chemistry , Radiation-Sensitizing Agents/radiation effects , SARS-CoV-2/isolation & purification , Temperature , Tissue Engineering/methods , Ultrasonic Waves , Viral Load/methods
10.
Biosens Bioelectron ; 200: 113909, 2022 Mar 15.
Article in English | MEDLINE | ID: covidwho-1670212

ABSTRACT

Coronavirus disease 2019 (COVID-19) has been recognized as a global pandemic outbreak, opening the most severe socio-economic crisis since World War II. Different scientific activities have been emerged in this global scenario, including the development of innovative analytical tools to measure nucleic acid, antibodies, and antigens in the nasopharyngeal swab, serum, and saliva for prompt identification of COVID-19 patients and to evaluate the immune response to the vaccine. The detection of SARS-CoV-2 in saliva remains a challenge for the lack of sufficient sensitivity. To address this issue, we developed a novel paper-based immunoassay using magnetic beads to support the immunological chain and 96-well wax-printed paper plate as a platform for color visualization by using a smartphone combined with Spotxel free-charge app. To assess the reliability of the measurement of SARS-CoV-2 in saliva, untreated saliva was used as a specimen and the calibration curve demonstrated a dynamic range up to 10 µg/mL, with a detection limit equal to 0.1 µg/mL. The effectiveness of this sustainable analytical tool in saliva was evaluated by comparing the data with the nasopharyngeal swab specimens sampled by the same patients and tested with Real-Time PCR reference method, founding 100% of agreement, even in the case of high Cycle Threshold (CT) numbers (low viral load). Furthermore, the positive saliva samples were characterized by the next-generation sequencing method, demonstrating the capability to detect the Delta variant, which is actually (July 2021) the most relevant variant of concern.


Subject(s)
Biosensing Techniques , COVID-19 , Colorimetry , Humans , Immunoassay , Magnetic Phenomena , Nasopharynx , Reproducibility of Results , SARS-CoV-2 , Saliva , Smartphone , Specimen Handling
11.
J Virol Methods ; 300: 114414, 2022 02.
Article in English | MEDLINE | ID: covidwho-1636596

ABSTRACT

COVID-19 has posed a worldwide public health challenge affecting millions of people in different countries. Rapid and efficient detection of SARS-CoV-2 is essential for pandemic control. Reverse Transcription quantitative PCR (RT-qPCR) of nasopharyngeal swabs is the gold standard method for the virus detection, but the high demand for tests has substantially increased the costs and reduced the availability of reagents, including genetic material purification kits. Thus, the present study aimed to compare two bead-based RNA extraction methods (an in-house and a commercial kit) from nasopharyngeal swabs and RT-qPCR detection of SARS-CoV-2. Twenty-five positive and five negative nasopharyngeal swab samples were subjected to extraction of nucleic acids using both methods in an automated platform. Both protocols revealed a high correlation between Cycle Quantifications (Cqs) (r = 0.99, p < 0.0001). In addition, the in-house kit was 89.5 % cheaper when compared to the mean cost of commercial RNA extraction kits. The results show that the in-house protocol is an affordable and reliable option for RNA extraction for SARS-CoV-2 detection from nasopharyngeal swabs.


Subject(s)
COVID-19 , COVID-19 Testing , Humans , Magnetic Phenomena , Nasopharynx , RNA, Viral/genetics , SARS-CoV-2 , Sensitivity and Specificity
12.
Water Res ; 212: 118112, 2022 Apr 01.
Article in English | MEDLINE | ID: covidwho-1636095

ABSTRACT

Viruses are present at low concentrations in wastewater; therefore, an effective method for concentrating virus particles is necessary for accurate wastewater-based epidemiology (WBE). We designed a novel approach to concentrate human and animal viruses from wastewater using porcine gastric mucin-conjugated magnetic beads (PGM-MBs). We systematically evaluated the performances of the PGM-MBs method (sensitivity, specificity, and robustness to environmental inhibitors) with six viral species, including Tulane virus (a surrogate for human norovirus), rotavirus, adenovirus, porcine coronavirus (transmissible gastroenteritis virus or TGEV), and two human coronaviruses (NL63 and SARS-CoV-2) in influent wastewater and raw sewage samples. We determined the multiplication factor (the ratio of genome concentration of the final solution to that of the initial solution) for the PGM-MBs method, which ranged from 1.3 to 64.0 depending on the viral species. Because the recovery efficiency was significantly higher when calculated with virus titers than it was with genome concentration, the PGM-MBs method could be an appropriate tool for assessing the risk to humans who are inadvertently exposed to wastewater contaminated with infectious viruses. Furthermore, PCR inhibitors were not concentrated by PGM-MBs, suggesting that this tool will be successful for use with environmental samples. In addition, the PGM-MBs method is cost-effective (0.5 USD/sample) and has a fast turnaround time (3 h from virus concentration to genome quantification). Thus, this method can be implemented in high throughput facilities. Because of its strong performance, intrinsic characteristics of targeting the infectious virus, robustness to wastewater, and adaptability to high throughput systems, the PGM-MBs method can be successfully applied to WBE and ultimately provides valuable public health information.


Subject(s)
COVID-19 , Viruses , Animals , Humans , Magnetic Phenomena , SARS-CoV-2 , Swine , Waste Water
13.
Talanta ; 241: 123243, 2022 May 01.
Article in English | MEDLINE | ID: covidwho-1633997

ABSTRACT

Viral diseases are the primary source of death, making a worldwide influence on healthcare, social, and economic development. Thus, diagnosis is the vital approach to the main aim of virus control and elimination. On the other hand, the prompt advancement of nanotechnology in the field of medicine possesses the probability of being beneficial to diagnose infections normally in labs as well as specifically. Nanoparticles are efficiently in use to make novel strategies because of permitting analysis at cellular in addition to the molecular scale. Henceforth, they assist towards pronounced progress concerning molecular analysis at the nanoscale. In recent times, magnetic nanoparticles conjugated through covalent bonds to bioanalytes for instance peptides, antibodies, nucleic acids, plus proteins are established like nanoprobes aimed at molecular recognition. These modified magnetic nanoparticles could offer a simple fast approach for extraction, purification, enrichment/concentration, besides viruses' recognition precisely also specifically. In consideration of the above, herein insight and outlook into the limitations of conventional methods and numerous roles played by magnetic nanoparticles to extract, purify, concentrate, and additionally in developing a diagnostic regime for viral outbreaks to combat viruses especially the ongoing novel coronavirus (COVID-19).


Subject(s)
COVID-19 , Viruses , Humans , Magnetic Phenomena , Magnetics , SARS-CoV-2 , Viruses/genetics
14.
Biosensors (Basel) ; 12(1)2021 Dec 23.
Article in English | MEDLINE | ID: covidwho-1581024

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
15.
Methods ; 203: 259-267, 2022 07.
Article in English | MEDLINE | ID: covidwho-1521610

ABSTRACT

The construction of a rapid, simple, and specific nucleic acid detection platform is of great significance to the control of the large-scale spread of infectious diseases. We have recently established a magnetic pull-down-assisted colorimetric method based on the CRISPR/Cas12a system (termed M-CDC), which effectively integrates the advantages of CRISPR/Cas12a, magnetic beads-based separation, and AuNP bioprobe to provide a simple and specific biosensing platform for nucleic acid assay. The M-CDC method is compatible with point-of-care testing and enables the detection of nucleic acid samples in less than an hour without relying on expensive and complex instruments. In this paper, step-by-step instructions for M-CDC assay, including recombinase polymerase amplification (RPA)/reverse transcription-polymerase chain reaction (RT-RPA) of DNA or RNA, Cas12a-mediated target recognition and cleavage, and subsequent magnetic beads-mediated colorimetric readouts are provided. In addition, the protocol for the expression and purification of Lachnospiraceae bacterium-Cas12a (LbCas12a) protein, the design and synthesis of high-efficient crRNA, and the preparation of AuNP bioprobe are also offered.


Subject(s)
CRISPR-Cas Systems , Nucleic Acid Amplification Techniques , CRISPR-Cas Systems/genetics , Centers for Disease Control and Prevention, U.S. , Colorimetry , DNA , Magnetic Phenomena , Nucleic Acid Amplification Techniques/methods , United States
16.
J Mol Diagn ; 23(12): 1680-1690, 2021 12.
Article in English | MEDLINE | ID: covidwho-1504354

ABSTRACT

Rapid and sensitive detection of human pathogens, such as the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), is an urgent and challenging task for clinical laboratories. Currently, the gold standard for SARS-CoV-2-specific RNA is based on quantitative RT-PCR (RT-qPCR), which relies on target amplification by Taq polymerase and uses a fluorescent resonance energy transfer-based hydrolysis probe. Although this method is accurate and specific, it is also time consuming. Here, a new molecular assay is described that combines a highly sensitive magnetic modulation biosensing (MMB) system, rapid thermal cycling, and a modified double-quenched hydrolysis probe. In vitro transcribed SARS-CoV-2 RNA targets spiked in PCR-grade water, were used to show that the calculated limit of detection of the MMB-based molecular assay was 1.6 copies per reaction. Testing 309 RNA extracts from 170 confirmed RT-qPCR SARS-CoV-2-negative individuals (30 of whom were positive for other respiratory viruses) and 139 RT-qPCR SARS-CoV-2-positive patients (CT ≤ 42) resulted in 97.8% sensitivity, 100% specificity, and 0% cross-reactivity. The total turnaround time of the MMB-based assay is 30 minutes, which is three to four times faster than a standard RT-qPCR. By adjusting the primers and the probe set, the platform can be easily adapted to detect most of the pathogens that are currently being diagnosed by RT-qPCR.


Subject(s)
COVID-19 Testing/methods , COVID-19/diagnosis , Molecular Diagnostic Techniques/methods , Humans , Magnetic Phenomena , Nucleic Acid Amplification Techniques/methods , RNA, Viral/genetics , Real-Time Polymerase Chain Reaction/methods , SARS-CoV-2/genetics , Sensitivity and Specificity
17.
J Virol Methods ; 299: 114339, 2022 01.
Article in English | MEDLINE | ID: covidwho-1472084

ABSTRACT

The COVID-19 pandemic has highlighted the need for generic reagents and flexible systems in diagnostic testing. Magnetic bead-based nucleic acid extraction protocols using 96-well plates on open liquid handlers are readily amenable to meet this need. Here, one such approach is rigorously optimized to minimize cross-well contamination while maintaining sensitivity.


Subject(s)
COVID-19 , Nucleic Acids , COVID-19 Testing , Humans , Indicators and Reagents , Magnetic Phenomena , Pandemics , RNA, Viral/genetics , SARS-CoV-2 , Sensitivity and Specificity
18.
Environ Pollut ; 291: 118191, 2021 Dec 15.
Article in English | MEDLINE | ID: covidwho-1427873

ABSTRACT

Between 9 March and 18 May 2020, strict lockdown measures were adopted in Italy for containing the COVID-19 pandemic: in Rome, despite vehicular traffic on average was more than halved, it was not observed a evident decrease of the airborne particulate matter (PM) concentrations, as assessed by air quality data. In this study, daily PM10 filters were collected from selected automated stations operated in Rome by the regional network of air quality monitoring: their magnetic properties - including magnetic susceptibility, hysteresis parameters and FORC (first order reversal curves) diagrams - were compared during and after the lockdown, for outlining the impact of the COVID-19 measures on airborne particulate matter. In urban traffic sites, the PM10 concentrations did not significantly change after the end of the lockdown, when vehicular traffic promptly returned to its usual levels; conversely, the average volume and mass magnetic susceptibilities approximately doubled, and the linear correlation between volume magnetic susceptibility and PM10 concentration became significant, pointing out the link between PM10 concentrations and the increasing levels of traffic-related magnetic emissions. Magnetite-like minerals, attributed to non-exhaust brakes emissions, dominated the magnetic fraction of PM10 near urban traffic sites, with natural magnetic components emerging in background sites and during exogenous dusts atmospheric events. Magnetic susceptibility constituted a fast and sensitive proxy of vehicular particulate emissions: the magnetic properties can play a relevant role in the source apportionment of PM10, especially when unsignificant variations in its concentration levels may mask important changes in the traffic-related magnetic fraction. As a further hint, increasing attention should be drawn to the reduction of brake wear emissions, that are overcoming by far fuel exhausts as the main particulate pollutant in traffic contexts.


Subject(s)
Air Pollutants , Air Pollution , COVID-19 , Air Pollutants/analysis , Air Pollution/analysis , Communicable Disease Control , Environmental Monitoring , Humans , Italy , Magnetic Phenomena , Pandemics , Particulate Matter/analysis , Rome , SARS-CoV-2 , Vehicle Emissions/analysis
19.
Angew Chem Int Ed Engl ; 60(21): 11884-11891, 2021 05 17.
Article in English | MEDLINE | ID: covidwho-1384108

ABSTRACT

2D NOESY plays a central role in structural NMR spectroscopy. We have recently discussed methods that rely on solvent-driven exchanges to enhance NOE correlations between exchangeable and non-exchangeable protons in nucleic acids. Such methods, however, fail when trying to establish connectivities within pools of labile protons. This study introduces an alternative that also enhances NOEs between such labile sites, based on encoding a priori selected peaks by selective saturations. The resulting selective magnetization transfer (SMT) experiment proves particularly useful for enhancing the imino-imino cross-peaks in RNAs, which is a first step in the NMR resolution of these structures. The origins of these enhancements are discussed, and their potential is demonstrated on RNA fragments derived from the genome of SARS-CoV-2, recorded with better sensitivity and an order of magnitude faster than conventional 2D counterparts.


Subject(s)
Nuclear Magnetic Resonance, Biomolecular/methods , Protons , RNA, Viral/analysis , SARS-CoV-2/chemistry , Magnetic Phenomena , RNA, Viral/chemistry
20.
J Am Chem Soc ; 143(13): 4942-4948, 2021 04 07.
Article in English | MEDLINE | ID: covidwho-1387161

ABSTRACT

Multidimensional NOESY experiments targeting correlations between exchangeable imino and amino protons provide valuable information about base pairing in nucleic acids. It has been recently shown that the sensitivity of homonuclear correlations involving RNA's labile imino protons can be significantly enhanced, by exploiting the repolarization brought about by solvent exchanges. Homonuclear correlations, however, are of limited spectral resolution, and usually incapable of tackling relatively large homopolymers with repeating structures like RNAs. This study presents a heteronuclear-resolved version of those NOESY experiments, in which magnetization transfers between the aqueous solvent and the nucleic acid protons are controlled by selecting specific chemical shift combinations of a coupled 1H-15N spin pair. This selective control effectively leads to a pseudo-3D version of HSQC-NOESY, but with cross-peaks enhanced by ∼2-5× as compared with conventional 2D NOESY counterparts. The enhanced signal sensitivity as well as access to both 15N-1H and 1H-1H NOESY dimensions can greatly facilitate RNA assignments and secondary structure determinations, as demonstrated here with the analysis of genome fragments derived from the SARS-CoV-2 virus.


Subject(s)
Magnetic Phenomena , Magnetic Resonance Spectroscopy , RNA, Viral/chemistry , SARS-CoV-2/genetics , Temperature
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