A novel approach to concentrate human and animal viruses from wastewater using receptors-conjugated magnetic beads

AbstractViruses are present at low concentrations in wastewater, and therefore an effective concentration of 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 concentrated over 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 became significantly higher when calculated based on virus titers than genome concentration, the PGM-MBs method could be an appropriate tool for assessing the risk due to wastewater contaminated with infectious enteric viruses. PCR inhibitors were not concentrated by PGM-MBs, suggesting this tool will be successful for use with environmental samples. The PGM-MBs method is cost-effective (0.43 USD/sample) and fast turnaround (3 hours from virus concentration to genome quantification), and thus this method can be implemented for high throughput facilities. Based on good performance, intrinsic characteristics of targeting the infectious virus, robustness to wastewater, and adaptability to high throughput systems, we are confident that the PGM-MBs method can be applied for successful WBE and ultimately provides valuable public health information.Graphical abstract

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Dry Heat as a Decontamination Method for N95 Respirator Reuse

10.26434/chemrxiv.12290621.v2 ◽

2020 ◽

Author(s):

Chamteut Oh ◽

Elbashir Araud ◽

Joseph V. Puthussery ◽

Hezi Bai ◽

Gemma G. Clark ◽

...

Keyword(s):

Heat Treatment ◽

Healthcare Providers ◽

Transmissible Gastroenteritis Virus ◽

Viable Option ◽

Dry Heat ◽

Tulane Virus ◽

Fit Testing ◽

Transmissible Gastroenteritis ◽

Dry Heat Treatment ◽

N95 Respirators

<div> <div> <p>A pandemic such as COVID-19 can cause a sudden depletion in the worldwide supply of respirators, forcing healthcare providers to reuse them. In this study, we systematically evaluated dry heat treatment as a viable option for the safe decontamination of N95 respirators (1860, 3M) before its reuse. We found that the dry heat generated by an electric cooker (100°C, 5% relative humidity, 50 min) effectively inactivated Tulane virus (>5.2-log<sub>10</sub> reduction), rotavirus (>6.6-log<sub>10</sub> reduction), adenovirus (>4.0-log<sub>10</sub> reduction), and transmissible gastroenteritis virus (>4.7-log<sub>10</sub> reduction). The respirator integrity (determined based on the particle filtration efficiency and quantitative fit testing) was not compromised after 20 cycles of 50-min dry heat treatment. Based on these results, we propose dry heat decontamination generated by an electric cooker (e.g., rice cookers, instant pots, ovens) to be an effective and accessible decontamination method for the safe reuse of N95 respirators.<br></p> </div> </div>

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A LITERATURE REVIEW OF NOVEL COVID-19 AND RELATED CORONAVIRUSES OUTBREAKS IN THE 1960S UNTIL 21ST CENTURY

EPRA International Journal of Multidisciplinary Research (IJMR) ◽

10.36713/epra5795 ◽

2020 ◽

pp. 353-366

Author(s):

Nurhafizul Abu Seri ◽

Anisah Jessica Lee

Keyword(s):

Hepatitis Virus ◽

Transmissible Gastroenteritis Virus ◽

The Public ◽

Turkey Coronavirus ◽

Baseline Information ◽

Transmissible Gastroenteritis ◽

Dry Climates ◽

Human Coronaviruses ◽

The 1960S ◽

Respiratory Coronavirus

Coronavirus (CoVs) is a large group of viruses known to affect birds and mammals including humans. This review aims to present the types of human coronavirus and animal coronavirus studied and displaying the relationships of these coronaviruses to weather, meteorology and climatology. Human coronaviruses (HCoV) in review are namely 229E, NL63, OC43 and HKU1 and other HCoV which are Severe Acute Respiratory Syndrome (SARS-CoV), Middle East Respiratory Syndrome (MERS-CoV) and COVID-19 (SARS-CoV-2). These HcoV originated as animal infections which then develop and ultimately transmitted to humans. CoV can also be found in animals which are Canine Respiratory Coronavirus (CRCoV), Murine Coronavirus Rat Hepatitis Virus (MHV), Transmissible Gastroenteritis Virus (TGEV), Bovine Coronavirus (BCoV), Feline Coronavirus (FCoV), Canine Coronavirus (CCoV) and Turkey Coronavirus (TCV). CoV has been found to last longer in the atmosphere at lower temperatures and lower relative humidity. Thus, some coronavirus outbreaks can mostly be attributed to cold and dry climates for more effective CoV transmission. Identified CoV are mostly active when the temperature is between 9 °C and 24 °C. Findings in this review can serve as knowledge and guidance for individuals, related organizations and governments to be prepared for the CoV threats that is currently occurring and that is likely to re-emerge in the coming years. It is also intended to provide useful baseline information for policymakers and the public. KEYWORDS: climatology, coronavirus (CoVs), human coronaviruses (HCoV), meteorology, weather

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Semi-Automated Library Preparation for High-Throughput DNA Sequencing Platforms

Journal of Biomedicine and Biotechnology ◽

10.1155/2010/617469 ◽

2010 ◽

Vol 2010 ◽

pp. 1-8 ◽

Cited By ~ 21

Author(s):

Eveline Farias-Hesson ◽

Jonathan Erikson ◽

Alexander Atkins ◽

Peidong Shen ◽

Ronald W. Davis ◽

...

Keyword(s):

Sample Preparation ◽

Dna Sequencing ◽

High Throughput ◽

Magnetic Beads ◽

Dna Barcode ◽

Cost Effective ◽

Dna Libraries ◽

Emulsion Pcr ◽

High Throughput Dna Sequencing ◽

Sequencing Platforms

Next-generation sequencing platforms are powerful technologies, providing gigabases of genetic information in a single run. An important prerequisite for high-throughput DNA sequencing is the development of robust and cost-effective preprocessing protocols for DNA sample library construction. Here we report the development of a semi-automated sample preparation protocol to produce adaptor-ligated fragment libraries. Using a liquid-handling robot in conjunction with Carboxy Terminated Magnetic Beads, we labeled each library sample using a unique 6 bp DNA barcode, which allowed multiplex sample processing and sequencing of 32 libraries in a single run using Applied Biosystems' SOLiD sequencer. We applied our semi-automated pipeline to targeted medical resequencing of nuclear candidate genes in individuals affected by mitochondrial disorders. This novel method is capable of preparing as much as 32 DNA libraries in 2.01 days (8-hour workday) for emulsion PCR/high throughput DNA sequencing, increasing sample preparation production by 8-fold.

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The Basis of Peracetic Acid Inactivation Mechanisms for Rotavirus and Tulane Virus under Conditions Relevant for Vegetable Sanitation

Applied and Environmental Microbiology ◽

10.1128/aem.01095-20 ◽

2020 ◽

Vol 86 (19) ◽

Author(s):

Miyu Fuzawa ◽

Hezi Bai ◽

Joanna L. Shisler ◽

Thanh H. Nguyen

Keyword(s):

Peracetic Acid ◽

Molecular Mechanisms ◽

Magnetic Beads ◽

Virus Titer ◽

Human Norovirus ◽

Porcine Gastric Mucin ◽

Genome Damage ◽

Versus Protein ◽

Receptor Interactions ◽

Tulane Virus

ABSTRACT We determined the disinfection efficacy and inactivation mechanisms of peracetic acid (PAA)-based sanitizer using pH values relevant for vegetable sanitation against rotavirus (RV) and Tulane virus (TV; a human norovirus surrogate). TV was significantly more resistant to PAA disinfection than RV: for a 2-log10 reduction of virus titer, RV required 1 mg/liter PAA for 3.5 min of exposure, while TV required 10 mg/liter PAA for 30 min. The higher resistance of TV can be explained, in part, by significantly more aggregation of TV in PAA solutions. The PAA mechanisms of virus inactivation were explored by quantifying (i) viral genome integrity and replication using reverse transcription-quantitative PCR (RT-qPCR) and (ii) virus-host receptor interactions using a cell-free binding assay with porcine gastric mucin conjugated with magnetic beads (PGM-MBs). We observed that PAA induced damage to both RV and TV genomes and also decreased virus-receptor interactions, with the latter suggesting that PAA damages viral proteins important for binding its host cell receptors. Importantly, the levels of genome-versus-protein damage induced by PAA were different for each virus. PAA inactivation correlated with higher levels of RV genome damage than of RV-receptor interactions. For PAA-treated TV, the opposite trends were observed. Thus, PAA inactivates each of these viruses via different molecular mechanisms. The findings presented here potentially contribute to the design of a robust sanitation strategy for RV and TV using PAA to prevent foodborne disease. IMPORTANCE In this study, we examined the inactivation mechanisms of peracetic acid (PAA), a sanitizer commonly used for postharvest vegetable washing, for two enteric viruses: Tulane virus (TV) as a human norovirus surrogate and rotavirus (RV). PAA disinfection mechanisms for RV were mainly due to genome damage. In contrast, PAA disinfection in TV was due to damage of the proteins important for binding to its host receptor. We also observed that PAA triggered aggregation of TV to a much greater extent than RV. These studies demonstrate that different viruses are inactivated via different PAA mechanisms. This information is important for designing an optimal sanitation practice for postharvest vegetable washing to minimize foodborne viral diseases.

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Strategy for Systematic Assembly of Large RNA and DNA Genomes: Transmissible Gastroenteritis Virus Model

Journal of Virology ◽

10.1128/jvi.74.22.10600-10611.2000 ◽

2000 ◽

Vol 74 (22) ◽

pp. 10600-10611 ◽

Cited By ~ 165

Author(s):

Boyd Yount ◽

Kristopher M. Curtis ◽

Ralph S. Baric

Keyword(s):

Full Length ◽

Host Cells ◽

Transmissible Gastroenteritis Virus ◽

Plaque Morphology ◽

Permissive Host ◽

Gastroenteritis Virus ◽

Novel Approach ◽

Transmissible Gastroenteritis ◽

Cdna Construct ◽

Rna And Dna

ABSTRACT A systematic method was developed to assemble functional full-length genomes of large RNA and DNA viruses. Coronaviruses contain the largest single-stranded positive-polarity RNA genome in nature. The ∼30-kb genome, coupled with regions of genomic instability, has hindered the development of a full-length infectious cDNA construct. We have assembled a full-length infectious construct of transmissible gastroenteritis virus (TGEV), an important pathogen in swine. Using a novel approach, six adjoining cDNA subclones that span the entire TGEV genome were isolated. Each clone was engineered with unique flanking interconnecting junctions which determine a precise systematic assembly with only the adjacent cDNA subclones, resulting in an intact TGEV cDNA construct of ∼28.5 kb in length. Transcripts derived from the full-length TGEV construct were infectious, and progeny virions were serially passaged in permissive host cells. Viral antigen production and subgenomic mRNA synthesis were evident during infection and throughout passage. Plaque-purified virus derived from the infectious construct replicated efficiently and displayed similar plaque morphology in permissive host cells. Host range phenotypes of the molecularly cloned and wild-type viruses were similar in cells of swine and feline origin. The recombinant viruses were sequenced across the unique interconnecting junctions, conclusively demonstrating the marker mutations and restriction sites that were engineered into the component clones. Full-length infectious constructs of TGEV will permit the precise genetic modification of the coronavirus genome. The method that we have designed to generate an infectious cDNA construct of TGEV could theoretically be used to precisely reconstruct microbial or eukaryotic genomes approaching several million base pairs in length.

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