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1.
Epidemiol Prev ; 44(5-6): 330-332, 2020.
Article in Italian | MEDLINE | ID: covidwho-2240354

ABSTRACT

Systematic reviews have shown a prevalence close to 20% of gastrointestinal symptoms in COVID-19 positive patients, with nearly 40% of patients shedding viral RNA in their faeces, even if it may not be infectious, possibly because of inactivation by colonic fluid.According to current evidence, this virus is primarily transmitted by respiratory droplets and contact routes, including contaminated surfaces. The virus is quite stable on stainless steel, being detected up to 48-72 hours after application. Therefore, some individuals can be infected touching common contaminated surfaces, such as bathroom taps. Taps can be underestimated critical points in the transmission chain of the infection. Indeed, just by turning the knob, people leave germs on it, especially after coughing over their hands, sneezing, and/or blowing their nose. After handwashing with soap, user take back their germs when turning the knob. Paradoxically, the following user collects the germs back on his/her fingers by implementing a preventive measure, maybe before putting food into the mouth or wearing contact lenses.The Italian National Institute of Health recommends to clean and disinfect high-touched surfaces, but it is unrealistic and inefficient to do so after each tap use. As an alternative, new toilets should install long elbow-levers - or at least short levers - provided that people are educated to close them with the forearm or the side of the hand. This is already a standard measure in hospitals, but it is particularly important also in high-risk communities, such as retirement homes and prisons. It would be important also in schools, in workplaces, and even in families, contributing to the prevention both of orofaecal and respiratory infections.In the meantime, people should be educated to close existing knobs with disposable paper towel wipes or with toilet paper sheets.


Subject(s)
Bathroom Equipment/virology , COVID-19/prevention & control , Fomites/virology , Hand Hygiene , Health Education , SARS-CoV-2/physiology , COVID-19/transmission , Equipment Contamination , Equipment Design , Feces/virology , Female , Humans , Italy , Male , SARS-CoV-2/isolation & purification , Touch
2.
Toxins (Basel) ; 12(4)2020 04 02.
Article in English | MEDLINE | ID: covidwho-1453289

ABSTRACT

Bacterial toxins play a key role in the pathogenesis of lung disease. Based on their structural and functional properties, they employ various strategies to modulate lung barrier function and to impair host defense in order to promote infection. Although in general, these toxins target common cellular signaling pathways and host compartments, toxin- and cell-specific effects have also been reported. Toxins can affect resident pulmonary cells involved in alveolar fluid clearance (AFC) and barrier function through impairing vectorial Na+ transport and through cytoskeletal collapse, as such, destroying cell-cell adhesions. The resulting loss of alveolar-capillary barrier integrity and fluid clearance capacity will induce capillary leak and foster edema formation, which will in turn impair gas exchange and endanger the survival of the host. Toxins modulate or neutralize protective host cell mechanisms of both the innate and adaptive immunity response during chronic infection. In particular, toxins can either recruit or kill central players of the lung's innate immune responses to pathogenic attacks, i.e., alveolar macrophages (AMs) and neutrophils. Pulmonary disorders resulting from these toxin actions include, e.g., acute lung injury (ALI), the acute respiratory syndrome (ARDS), and severe pneumonia. When acute infection converts to persistence, i.e., colonization and chronic infection, lung diseases, such as bronchitis, chronic obstructive pulmonary disease (COPD), and cystic fibrosis (CF) can arise. The aim of this review is to discuss the impact of bacterial toxins in the lungs and the resulting outcomes for pathogenesis, their roles in promoting bacterial dissemination, and bacterial survival in disease progression.


Subject(s)
Bacteria/pathogenicity , Bacterial Infections/microbiology , Bacterial Toxins/metabolism , Lung/microbiology , Respiratory Tract Infections/microbiology , Adaptive Immunity , Animals , Bacteria/immunology , Bacteria/metabolism , Bacterial Infections/immunology , Bacterial Infections/metabolism , Bacterial Infections/pathology , Disease Progression , Host-Pathogen Interactions , Humans , Immunity, Innate , Lung/immunology , Lung/metabolism , Lung/pathology , Respiratory Tract Infections/immunology , Respiratory Tract Infections/metabolism , Respiratory Tract Infections/pathology , Signal Transduction
3.
Int J Environ Res Public Health ; 17(23)2020 12 05.
Article in English | MEDLINE | ID: covidwho-1389355

ABSTRACT

Our research aimed to review the potential risk of infection by SARS-CoV-2. We used an excerpt of a data set generated in May 2020 for reviewing the SARS-CoV-2 prevention concept of orchestras, singers and actors. People were sampled for droplet release for one-hour activities using a Grimm spectrometer covering a spectrum of 1 to 32 µm diameter. We estimated the number of "quanta" in the exhaled liquid from viral concentrations of 106 to 1011/mL, based on the Human Infective Dose 50 of 218 viral particles. We employed the Wells-Riley equation to estimate the risk of infection in typical meeting rooms for a one-hour meeting of 2, 4 and 6 people observing a 2 m distance. The four participating adults released a mean of 1.28 nLm3 while breathing, 1.68 nL/m3 while speaking normally, and two adults released a mean of 4.44 nL/m3 while talking with a raised voice. The combination of 50% breathing, 45% talking normally and 5% speaking with a raised voice increased the risk of infection above 5% for a one-hour meeting of two people. The result is based on 6 quanta released, corresponding to an initial virus concentration of 1000/nL (109/mL) in the fluid of the upper respiratory tract. Our data confirm the importance of using facemasks in combination with other measures to prevent transmission of SARS-CoV-2 at the workplace.


Subject(s)
Aerosols , Air Microbiology , COVID-19/transmission , Speech , Healthy Volunteers , Humans , Pandemics , SARS-CoV-2
5.
Am J Respir Cell Mol Biol ; 64(6): 677-686, 2021 06.
Article in English | MEDLINE | ID: covidwho-1259048

ABSTRACT

There is an urgent need for new drugs for patients with acute respiratory distress syndrome (ARDS), including those with coronavirus disease (COVID-19). ARDS in influenza-infected mice is associated with reduced concentrations of liponucleotides (essential precursors for de novo phospholipid synthesis) in alveolar type II (ATII) epithelial cells. Because surfactant phospholipid synthesis is a primary function of ATII cells, we hypothesized that disrupting this process could contribute significantly to the pathogenesis of influenza-induced ARDS. The goal of this study was to determine whether parenteral liponucleotide supplementation can attenuate ARDS. C57BL/6 mice inoculated intranasally with 10,000 plaque-forming units/mouse of H1N1 influenza A/WSN/33 virus were treated with CDP (cytidine 5'-diphospho)-choline (100 µg/mouse i.p.) ± CDP -diacylglycerol 16:0/16:0 (10 µg/mouse i.p.) once daily from 1 to 5 days after inoculation (to model postexposure influenza prophylaxis) or as a single dose on Day 5 (to model treatment of patients with ongoing influenza-induced ARDS). Daily postexposure prophylaxis with CDP-choline attenuated influenza-induced hypoxemia, pulmonary edema, alterations in lung mechanics, impairment of alveolar fluid clearance, and pulmonary inflammation without altering viral replication. These effects were not recapitulated by the daily administration of CTP (cytidine triphosphate) and/or choline. Daily coadministration of CDP-diacylglycerol significantly enhanced the beneficial effects of CDP-choline and also modified the ATII cell lipidome, reversing the infection-induced decrease in phosphatidylcholine and increasing concentrations of most other lipid classes in ATII cells. Single-dose treatment with both liponucleotides at 5 days after inoculation also attenuated hypoxemia, altered lung mechanics, and inflammation. Overall, our data show that liponucleotides act rapidly to reduce disease severity in mice with severe influenza-induced ARDS.


Subject(s)
Alveolar Epithelial Cells/metabolism , Cytidine Diphosphate Choline/pharmacology , Cytidine Diphosphate Diglycerides/pharmacology , Influenza A Virus, H1N1 Subtype/metabolism , Orthomyxoviridae Infections/drug therapy , Respiratory Distress Syndrome/prevention & control , Alveolar Epithelial Cells/pathology , Alveolar Epithelial Cells/virology , Animals , COVID-19/pathology , Mice , Orthomyxoviridae Infections/complications , Orthomyxoviridae Infections/metabolism , Orthomyxoviridae Infections/pathology , Respiratory Distress Syndrome/etiology , Respiratory Distress Syndrome/metabolism , Respiratory Distress Syndrome/pathology , SARS-CoV-2/metabolism , COVID-19 Drug Treatment
6.
MRS Adv ; 6(18): 463-466, 2021.
Article in English | MEDLINE | ID: covidwho-1256107

ABSTRACT

ABSTRACT: With the Covid-19-based global pandemic that started in the beginning of 2020, the vital importance of accelerated, reliable and affordable virus testing systems has once again become clearer. Besides, we all learned very well that the disposable biochips, to be used in these in vitro diagnostic (IVD) testing systems, supposed to be produced in large amounts in a very short time to be widely available for the use of humanity to save more and more lives. That is why; roll-to-roll (R2R) polymer structuring manners offer such large quantities for the production of in vitro biochips. Our technology, based on R2R UV nanoimprint lithography (UV-NIL), has superior features. Via our pilot line, robust 7500 biochip components per 100 meter of a flexible, polymer foil coated with a UV curable photo-resin (i.e., parts with capillary fluidic channels or optical structures for IVDs) can be generated. This study shows an example of a prototype of a R2R UV-NIL generated chip: a foil, capillary flow-based IVD biochip for multiplexed DNA detection purposes (i.e., a Lab-on-a-Foil device). The biochip performance was further increased dramatically by integrating UV-NIL produced retro-reflective microstructures, which reflects the light back, to its design to enhance optical signal detection in a commercial IVD device, detecting DNA on a chemiluminescent-reaction basis.

7.
Indian J Ophthalmol ; 69(6): 1592-1597, 2021 Jun.
Article in English | MEDLINE | ID: covidwho-1236859

ABSTRACT

Since the outbreak of respiratory coronavirus disease (COVID-19) caused by the coronavirus SARS-CoV-2, there is an ongoing discussion about whether the virus could be transmitted through corneal transplantation from donor to recipient. The purpose of this review was to summarize the current knowledge in the scientific community to provide aid in risk evaluation for potential virus transfer by corneal transplants. Literature was searched in PubMed.gov for relevant articles on coronavirus in conjunction with cornea processing, cornea transplantation and eye banking. Further, guidelines of health authorities and eye banking associations were reviewed. Studies have shown that SARS-CoV-2 RNA can be detected in ocular swabs and/or fluid of patients with COVID-19. However, the risk of SARS-CoV-2 virus transmission through these ocular tissues or fluid of patients is judged differently. To date, per literature and official guidelines, no evidence of viable virus in ocular tissue and no cases of transmission of SARS-CoV-2 via tissue preparations have been reported.


Subject(s)
COVID-19 , SARS-CoV-2 , Cornea , Eye Banks , Humans , RNA, Viral
8.
Cell Stem Cell ; 28(7): 1205-1220.e7, 2021 07 01.
Article in English | MEDLINE | ID: covidwho-1230788

ABSTRACT

The SARS-CoV-2 pandemic has caused unparalleled disruption of global behavior and significant loss of life. To minimize SARS-CoV-2 spread, understanding the mechanisms of infection from all possible routes of entry is essential. While aerosol transmission is thought to be the primary route of spread, viral particles have been detected in ocular fluid, suggesting that the eye may be a vulnerable point of viral entry. To this end, we confirmed SARS-CoV-2 entry factor and antigen expression in post-mortem COVID-19 patient ocular surface tissue and observed productive viral replication in cadaver samples and eye organoid cultures, most notably in limbal regions. Transcriptional analysis of ex vivo infected ocular surface cells and hESC-derived eye cultures revealed robust induction of NF-κB in infected cells as well as diminished type I/III interferon signaling. Together these data suggest that the eye can be directly infected by SARS-CoV-2 and implicate limbus as a portal for viral entry.


Subject(s)
COVID-19 , Human Embryonic Stem Cells , Adult , Epithelium , Humans , Pandemics , SARS-CoV-2
9.
Nanomedicine (Lond) ; 16(14): 1187-1202, 2021 06.
Article in English | MEDLINE | ID: covidwho-1226939

ABSTRACT

Aim: To formulate an aerosolized nanoliposomal carrier for remdesivir (AL-Rem) against coronavirus disease 2019. Methods: AL-Rem was prepared using modified hydration technique. Cytotoxicity in lung adenocarcinoma cells, stability and aerodynamic characteristics of developed liposomes were evaluated. Results: AL-Rem showed high encapsulation efficiency of 99.79%, with hydrodynamic diameter of 71.46 ± 1.35 nm and surface charge of -32 mV. AL-Rem demonstrated minimal cytotoxicity in A549 cells and retained monolayer integrity of Calu-3 cells. AL-Rem showed sustained release, with complete drug release obtained within 50 h in simulated lung fluid. Long-term stability indicated >90% drug recovery at 4°C. Desirable aerosol performance, with mass median aerodynamic diameter of 4.56 ± 0.55 and fine particle fraction of 74.40 ± 2.96%, confirmed successful nebulization of AL-Rem. Conclusion: AL-Rem represents an effective alternative for coronavirus disease 2019 treatment.


Lay abstract Remdesivir is one of the first drugs approved for the treatment of coronavirus disease 2019. Currently, it is administered via an injection into the bloodstream. This means that the drug circulates around the entire body and only a limited amount reaches the diseased site ­ the lungs. Frequent dosing is therefore required, which needs expert personnel and multiple hospital visits and can result in serious side effects. In this study, the authors developed specialized, nanosized particles containing the drug remdesivir that can be administered directly into the lungs. This could drastically minimize side effects, enhance efficacy and allow easy self-administration at home. The results of the study are promising but require additional investigation.


Subject(s)
Adenosine Monophosphate/analogs & derivatives , Alanine/analogs & derivatives , COVID-19 Drug Treatment , Drug Carriers , A549 Cells , Adenosine Monophosphate/administration & dosage , Administration, Inhalation , Aerosols , Alanine/administration & dosage , Delayed-Action Preparations , Drug Liberation , Humans , Liposomes , Nanoparticles , Particle Size
10.
J Appl Microbiol ; 131(5): 2567-2578, 2021 Nov.
Article in English | MEDLINE | ID: covidwho-1218121

ABSTRACT

AIMS: Assess the feasibility of using light from artificial sun lamps to decontaminate N95 filtering facepiece respirators (FFRs) contaminated with SARS-CoV-2. METHODS AND RESULTS: FFR coupons or whole FFRs contaminated with 5 log10 TCID50 (target concentration) SARS-CoV-2 in culture media, simulated saliva, or simulated lung fluid were dried for 1-2 h, then exposed to light from tanning and horticulture lamps to assess decontamination. Exposed coupons and whole FFRs showed SARS-CoV-2 inactivation for all matrices tested. Furthermore, FFRs still met performance specifications after five decontamination cycles. CONCLUSIONS: It is feasible that artificial sunlight from these sun lamps can be used to decontaminate FFRs provided the UV dose is sufficient and the light is unobstructed. Furthermore, decontamination can be performed up to five times without degrading FFR performance. SIGNIFICANCE AND IMPACT OF THE STUDY: This research shows a proof of principle that artificial sun lamps may be an option to decontaminate SARS-CoV-2 on N95 FFRs. UV doses required for inactivation to levels below detection ranged from 4 to 37·8 J cm-2 depending on the light source, virus matrix and FFR type.


Subject(s)
COVID-19 , Equipment Reuse , Decontamination , Humans , N95 Respirators , SARS-CoV-2
11.
Int Immunopharmacol ; 97: 107694, 2021 Aug.
Article in English | MEDLINE | ID: covidwho-1209540

ABSTRACT

In late 2019, a novel coronavirus (SARS-CoV-2) emerged in Wuhan city, Hubei province, China. Rapidly escalated into a worldwide pandemic, it has caused an unprecedented and devastating situation on the global public health and society economy. The severity of recent coronavirus disease, abbreviated to COVID-19, seems to be mostly associated with the patients' immune response. In this vein, mesenchymal stromal/stem cells (MSCs) have been suggested as a worth-considering option against COVID-19 as their therapeutic properties are mainly displayed in immunomodulation and anti-inflammatory effects. Indeed, administration of MSCs can attenuate cytokine storm and enhance alveolar fluid clearance, endothelial recovery, and anti-fibrotic regeneration. Despite advantages attributed to MSCs application in lung injuries, there are still several issues __foremost probability of malignant transformation and incidence of MSCs-related coagulopathy__ which should be resolved for the successful application of MSC therapy in COVID-19. In the present study, we review the historical evidence of successful use of MSCs and MSC-derived extracellular vesicles (EVs) in the treatment of acute respiratory distress syndrome (ARDS). We also take a look at MSCs mechanisms of action in the treatment of viral infections, and then through studying both the dark and bright sides of this approach, we provide a thorough discussion if MSC therapy might be a promising therapeutic approach in COVID-19 patients.


Subject(s)
COVID-19/therapy , Extracellular Vesicles/immunology , Mesenchymal Stem Cell Transplantation/methods , Mesenchymal Stem Cells/immunology , Respiratory Distress Syndrome/therapy , Anti-Inflammatory Agents/immunology , Anti-Inflammatory Agents/therapeutic use , COVID-19/complications , Humans , Respiratory Distress Syndrome/etiology
12.
Multimed Syst ; 28(4): 1223-1237, 2022.
Article in English | MEDLINE | ID: covidwho-1156947

ABSTRACT

Coronavirus is a fatal disease that affects mammals and birds. Usually, this virus spreads in humans through aerial precipitation of any fluid secreted from the infected entity's body part. This type of virus is fatal than other unpremeditated viruses. Meanwhile, another class of coronavirus was developed in December 2019, named Novel Coronavirus (2019-nCoV), first seen in Wuhan, China. From January 23, 2020, the number of affected individuals from this virus rapidly increased in Wuhan and other countries. This research proposes a system for classifying and analyzing the predictions obtained from symptoms of this virus. The proposed system aims to determine those attributes that help in the early detection of Coronavirus Disease (COVID-19) using the Adaptive Neuro-Fuzzy Inference System (ANFIS). This work computes the accuracy of different machine learning classifiers and selects the best classifier for COVID-19 detection based on comparative analysis. ANFIS is used to model and control ill-defined and uncertain systems to predict this globally spread disease's risk factor. COVID-19 dataset is classified using Support Vector Machine (SVM) because it achieved the highest accuracy of 100% among all classifiers. Furthermore, the ANFIS model is implemented on this classified dataset, which results in an 80% risk prediction for COVID-19.

13.
Dig Endosc ; 34(1): 96-104, 2022 Jan.
Article in English | MEDLINE | ID: covidwho-1132881

ABSTRACT

OBJECTIVES: Gastrointestinal endoscopy (GIE) is useful for the early detection and treatment of many diseases; however, GIE is considered a high-risk procedure in the coronavirus disease 2019 (COVID-19) pandemic era. This study aimed to explore the rate of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) positivity in saliva and gastrointestinal fluids to which endoscopy medical staff are exposed. METHODS: The study was a single-center cross-sectional study. From June 1 to July 31, 2020, all patients who underwent GIE at Yokohama City University Hospital were registered. All patients provided 3 mL of saliva. For upper GIE, 10 mL of gastric fluid was collected through the endoscope. For lower GIE, 10 mL of intestinal fluid was collected through the endoscope. The primary outcome was the positive rate of SARS-CoV-2 in saliva and gastrointestinal fluids. We also analyzed serum-specific antibodies for SARS-CoV-2 and patients' background information. RESULTS: A total of 783 samples (560 upper GIE and 223 lower GIE samples) were analyzed. Polymerase chain reaction (PCR) on saliva samples did not show any positive results in either upper or lower GIE samples. However, 2.0% (16/783) of gastrointestinal fluid samples tested positive for SARS-CoV-2. No significant differences in age, sex, purpose of endoscopy, medication, or rate of antibody test positivity were found between PCR positive and PCR negative cases. CONCLUSIONS: Asymptomatic patients, even those with no detectable virus in their saliva, had SARS-CoV-2 in their gastrointestinal tract. Endoscopy medical staff should be aware of infection when performing procedures. The study was registered as UMIN000040587.


Subject(s)
COVID-19 , SARS-CoV-2 , Cross-Sectional Studies , Endoscopy, Gastrointestinal , Humans , Japan/epidemiology , Prevalence , Prospective Studies , Saliva
14.
Sci Rep ; 11(1): 5448, 2021 03 09.
Article in English | MEDLINE | ID: covidwho-1125902

ABSTRACT

To safely re-open economies and prevent future outbreaks, rapid, frequent, point-of-need, SARS-CoV-2 diagnostic testing is necessary. However, existing field-deployable COVID-19 testing methods require the use of uncomfortable swabs and trained providers in PPE, while saliva-based methods must be transported to high complexity laboratories for testing. Here, we report the development and clinical validation of High-Performance Loop-mediated isothermal Amplification (HP-LAMP), a rapid, saliva-based, SARS-CoV-2 test with a limit of detection of 1.4 copies of virus per µl of saliva and a sensitivity and specificity with clinical samples of > 96%, on par with traditional RT-PCR based methods using swabs, but can deliver results using only a single fluid transfer step and simple heat block. Testing of 120 patient samples in 40 pools comprised of 5 patient samples each with either all negative or a single positive patient sample was 100% accurate. Thus, HP-LAMP may enable rapid and accurate results in the field using saliva, without need of a high-complexity laboratory.


Subject(s)
COVID-19/diagnosis , SARS-CoV-2/genetics , Saliva/virology , COVID-19/virology , COVID-19 Nucleic Acid Testing , Humans , Limit of Detection , Molecular Diagnostic Techniques , Nasopharynx/virology , Nucleic Acid Amplification Techniques , RNA, Viral/metabolism , SARS-CoV-2/isolation & purification , Sensitivity and Specificity , Temperature
15.
J Clin Med ; 10(6)2021 Mar 10.
Article in English | MEDLINE | ID: covidwho-1125241

ABSTRACT

Knowledge about the detection potential and detection rates of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in various body fluids and sites is important for dentists since they, directly or indirectly, deal with many of these fluids/sites in their daily practices. In this study, we attempt to review the latest evidence and meta-analysis studies regarding the detection rate of SARS-CoV-2 in different body specimens and sites as well as the characteristics of these sample. The presence/detection of SARS-CoV-2 viral biomolecules (nucleic acid, antigens, antibody) in different clinical specimens depends greatly on the specimen type and timing of collection. These specimens/sites include nasopharynx, oropharynx, nose, saliva, sputum, bronchoalveolar lavage, stool, urine, ocular fluid, serum, plasma and whole blood. The relative detection rate of SARS-CoV-2 viral biomolecules in each of these specimens/sites is reviewed in detail within the text. The infectious potential of these specimens depends mainly on the time of specimen collection and the presence of live replicating viral particles.

16.
Am J Physiol Lung Cell Mol Physiol ; 320(6): L1186-L1193, 2021 06 01.
Article in English | MEDLINE | ID: covidwho-1124630

ABSTRACT

A significant number of patients with coronavirus disease 2019 (COVID-19) develop acute respiratory distress syndrome (ARDS) that is associated with a poor outcome. The molecular mechanisms driving failure of the alveolar barrier upon severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection remain incompletely understood. The Na,K-ATPase is an adhesion molecule and a plasma membrane transporter that is critically required for proper alveolar epithelial function by both promoting barrier integrity and resolution of excess alveolar fluid, thus enabling appropriate gas exchange. However, numerous SARS-CoV-2-mediated and COVID-19-related signals directly or indirectly impair the function of the Na,K-ATPase, thereby potentially contributing to disease progression. In this Perspective, we highlight some of the putative mechanisms of SARS-CoV-2-driven dysfunction of the Na,K-ATPase, focusing on expression, maturation, and trafficking of the transporter. A therapeutic mean to selectively inhibit the maladaptive signals that impair the Na,K-ATPase upon SARS-CoV-2 infection might be effective in reestablishing the alveolar epithelial barrier and promoting alveolar fluid clearance and thus advantageous in patients with COVID-19-associated ARDS.


Subject(s)
COVID-19/pathology , Pulmonary Alveoli/pathology , Severe Acute Respiratory Syndrome/pathology , Sodium-Potassium-Exchanging ATPase/metabolism , Tight Junctions/pathology , Biological Transport/physiology , Humans , Pulmonary Edema/pathology , SARS-CoV-2
17.
Biochip J ; 15(1): 100-108, 2021.
Article in English | MEDLINE | ID: covidwho-1092859

ABSTRACT

Anti-severe acute respiratory syndrome coronavirus 2 (anti-SARS-CoV-2) nucleoprotein (NP) antibodies were isolated from pig sera using human SARS-CoV-2 NP-immobilized magnetic beads. The binding properties of the isolated antibodies against SARS-CoV-2 NP were tested via flow cytometry using SARS-CoV-2 NP-immobilized magnetic beads. A competitive immunoassay was developed for detecting SARS-CoV-2 NP as well as SARS-CoV-2 in the culture fluid using magnetic beads with immobilized anti-SARS-CoV-2 NP antibodies. Selectivity tests were carried out during the competitive immunoassay for SARS-CoV, MERS-CoV, and CoV strain 229E in the culture fluid.

18.
Acta Clin Belg ; 77(2): 368-376, 2022 Apr.
Article in English | MEDLINE | ID: covidwho-1081271

ABSTRACT

INTRODUCTION: We present the results of the COVID-19 rule-out protocol at Ghent University Hospital, a step-wise testing approach which included repeat NFS SARS-CoV-2 rRT-PCR, respiratory multiplex RT-PCR, low-dose chest CT and bronchoscopy with BAL to confirm or rule-out SARS-CoV-2 infection in patients admitted with symptoms suggestive of COVID-19. RESULTS: Between 19 March 2020 and 30 April 2020, 455 non-critically ill patients with symptoms suspect for COVID-19 were admitted. The initial NFS for SARS-CoV-2 rRT-PCR yielded 66.9%, the second NFS 25.4% and bronchoscopy with BAL 5.9% of total COVID-19 diagnoses. In the BAL fluid, other respiratory pathogens were detected in 65% (13/20) of the COVID-19 negative patients and only in 1/7 COVID-19 positive patients. Retrospective antibody testing at the time around BAL sampling showed a positive IgA or IgG in 42.9 % of the COVID-19 positive and 10.5% of the COVID-19 negative group. Follow-up serology showed 100% COVID-19 positivity in the COVID-19 positive group and 100% IgG negativity in the COVID-19 negative group. CONCLUSION: In our experience, bronchoscopy with BAL can have an added value to rule-in or rule-out COVID-19 in patients with clinical and radiographical high-likelihood of COVID-19 and repeated negative NFS testing. Furthermore, culture and respiratory multiplex PCR on BAL fluid can aid to identify alternative microbial etiological agents in this group. Retrospective analysis of antibody development in this selected group of patients suggests that the implementation of serological assays in the routine testing protocol will decrease the need for invasive procedures like bronchoscopy.


Subject(s)
COVID-19 , Bronchoscopy , COVID-19/diagnosis , Humans , Retrospective Studies , SARS-CoV-2 , Tomography, X-Ray Computed
19.
Cornea ; 40(3): 342-347, 2021 Mar 01.
Article in English | MEDLINE | ID: covidwho-1066449

ABSTRACT

PURPOSE: To examine corneal tissue for severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) positivity regarding implications for tissue procurement, processing, corneal transplantation, and ocular surgery on healthy patients. We performed quantitative reverse transcription-polymerase chain reaction qRT-PCR-testing for SARS-CoV-2 RNA on corneal stroma and endothelium, bulbar conjunctiva, conjunctival fluid swabs, anterior chamber fluid, and corneal epithelium of coronavirus disease 2019 (COVID-19) postmortem donors. METHODS: Included in this study were 10 bulbi of 5 COVID-19 patients who died because of respiratory insufficiency. Informed consent and institutional review board approval was obtained before this study (241/2020BO2). SARS-CoV-2 was detected by using a pharyngeal swab and bronchoalveolar lavage. Tissue procurement and tissue preparation were performed with personal protective equipment (PPE) and the necessary protective measures. qRT-PCR-testing was performed for each of the abovementioned tissues and intraocular fluids. RESULTS: The qRT-PCRs yielded no viral RNA in the following ocular tissues and intraocular fluid: corneal stroma and endothelium, bulbar-limbal conjunctiva, conjunctival fluid swabs, anterior chamber fluid, and corneal epithelium. CONCLUSIONS: In this study, no SARS-CoV-2-RNA was detected in conjunctiva, anterior chamber fluid, and corneal tissues (endothelium, stroma, and epithelium) of COVID-19 donors. This implicates that the risk for SARS-CoV-2 infection using corneal or conjunctival tissue is very low. However, further studies on a higher number of COVID-19 patients are necessary to confirm these results. This might be of high importance for donor tissue procurement, processing, and corneal transplantation.


Subject(s)
Aqueous Humor/virology , COVID-19/diagnosis , Conjunctiva/virology , Cornea/virology , Eye Infections, Viral/diagnosis , RNA, Viral/genetics , SARS-CoV-2/genetics , Aged , Aged, 80 and over , COVID-19/genetics , COVID-19/virology , COVID-19 Nucleic Acid Testing , Corneal Diseases/diagnosis , Corneal Diseases/genetics , Corneal Diseases/virology , Eye Banks , Eye Infections, Viral/genetics , Eye Infections, Viral/virology , Female , Humans , Male , Tissue Donors , Tissue and Organ Procurement
20.
PLoS One ; 16(1): e0245924, 2021.
Article in English | MEDLINE | ID: covidwho-1042604

ABSTRACT

SARS-CoV-2 is a single stranded RNA (ssRNA) virus and contains GU-rich sequences distributed abundantly in the genome. In COVID-19, the infection and immune hyperactivation causes accumulation of inflammatory immune cells, blood clots, and protein aggregates in lung fluid, increased lung alveolar wall thickness, and upregulation of serum cytokine levels. A serum protein called serum amyloid P (SAP) has a calming effect on the innate immune system and shows efficacy as a therapeutic for fibrosis in animal models and clinical trials. Here we show that aspiration of the GU-rich ssRNA oligonucleotide ORN06 into mouse lungs induces all of the above COVID-19-like symptoms. Men tend to have more severe COVID-19 symptoms than women, and in the aspirated ORN06 model, male mice tended to have more severe symptoms than female mice. Intraperitoneal injections of SAP starting from day 1 post ORN06 aspiration attenuated the ORN06-induced increase in the number of inflammatory cells and formation of clot-like aggregates in the mouse lung fluid, reduced ORN06-increased alveolar wall thickness and accumulation of exudates in the alveolar airspace, and attenuated an ORN06-induced upregulation of the inflammatory cytokines IL-1ß, IL-6, IL-12p70, IL-23, and IL-27 in serum. SAP also reduced D-dimer levels in the lung fluid. In human peripheral blood mononuclear cells, SAP attenuated ORN06-induced extracellular accumulation of IL-6. Together, these results suggest that aspiration of ORN06 is a simple model for both COVID-19 as well as cytokine storm in general, and that SAP is a potential therapeutic for diseases with COVID-19-like symptoms and/or a cytokine storm.


Subject(s)
COVID-19 Drug Treatment , Cytokine Release Syndrome/drug therapy , Pneumonia/drug therapy , Serum Amyloid P-Component/therapeutic use , Animals , COVID-19/complications , COVID-19/pathology , Cytokine Release Syndrome/complications , Cytokine Release Syndrome/pathology , Disease Models, Animal , Female , Humans , Lung/drug effects , Lung/pathology , Male , Mice , Mice, Inbred C57BL , Pneumonia/complications , Pneumonia/pathology , Serum Amyloid P-Component/administration & dosage
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