The Characteristics of Spike Glycoprotein Gene of Swine Acute Diarrhea Syndrome Coronavirus Strain CH/FJWT/2018 Isolated in China

Swine acute diarrhea syndrome (SADS) is a highly contagious infectious disease characterized by acute vomiting and watery diarrhea in neonatal piglets. The causative agent for SADS is the swine acute diarrhea syndrome coronavirus (SADS-CoV), an alphacoronavirus in the family Coronaviridae. Currently, SADS-CoV was identified only in Guangdong and Fujian provinces of China, not in any other regions or countries in the world. To explore the genetic diversity of SADS-CoV isolates, herein we comparatively analyzed 44 full-length genomes of viruses isolated in Guangdong and Fujian provinces during 2017–2019. The spike glycoprotein gene of SADS-CoV strain CH/FJWT/2018 isolated in Fujian province is distinct from that of other viral isolates in either spike glycoprotein gene-based phylogenetic analysis or whole genome-based gene similarity analysis. Moreover, at least 7 predicted linear B cell epitopes in the spike glycoprotein of CH/FJWT/2018 would be affected by amino acid variations when compared with a representative virus isolated in Guangdong province. The spike glycoprotein of coronaviruses determines viral host range and tissue tropism during virus infection via specific interactions with the cellular receptor and also plays critical roles in eliciting the production of neutralizing antibodies. Since SADS-CoVs have a broad cell tropism, the results in this report further emphasize that the spike glycoprotein gene is a pivotal target in the surveillance of SADS-CoV.

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Peculiarities of PED virus pathogenesis in neonatal non-immune piglets on Ukraine farms

Regulatory Mechanisms in Biosystems ◽

10.15421/021878 ◽

2018 ◽

Vol 9 (4) ◽

pp. 522-528

Author(s):

D. N. Masiuk ◽

V. S. Nedzvetsky ◽

A. I. Sosnztskyi ◽

A. V. Kokariev ◽

A. I. Zavhorodnii

Keyword(s):

Small Intestine ◽

Acute Infection ◽

Clinical Signs ◽

Infection Process ◽

Watery Diarrhea ◽

Agricultural Enterprises ◽

Neonatal Piglets ◽

Initial Stage ◽

Show Evidence ◽

Diarrhea Syndrome

The article presents the results of study of the immune and tissue features of pathogenesis of PED virus in neonatal piglets in Ukraine. Clinical and epizootological analysis was performed in three separate agricultural enterprises with adverse epidemiological situation with PED virus. Blood samples and fragments of the small intestine of PEDv infected piglets were used to study the characteristics of the pathogenesis of PED virus. It has been determined that the pathogenesis of super-acute infection by the PED virus in 1–7 day old non-immune piglets proceeds according to the classic type of infectious process with a relay transmission of the pathogen. Moreover, diarrhea syndrome has induced the development of irreversible and lethal pathophysiological lesions by replication the PED virus in enterocytes of infected piglets. Pathogenetic changes observed in PEDv infected piglets were combined with the following components: acute inflammatory-necrotic process in the tissue of the intestine, destruction and desquamation of the epithelium of the small intestine. The infection process on day 3–5 of manifestation of clinical signs of PED virus is accompanied with an increase in the number of erythrocytes, leukocytes and monocytes, hemoglobin, hematocrit level and aminotransferase activity. In addition, PED virus infection induced an increase in IgM levels and the number of phagocytizing leukocytes with increased aggressiveness and digestive power. The duration of the release of PED virus with feces is 26–37 days after cessation of watery diarrhea. The concentration of PEDv in the feces of convalescent piglets ranged from 120 to 1700 equivalent genome in 1 g of tissue. Parasite coenotic associations of the microbiocenosis of the digestive tract of PED virus infected piglets did not have time to form into stable epitopes. There was a constant quantitative prevalence of transient microbiota and a decrease in the share of probiotic microorganisms. The intestinal microflora consists of random transient microbiota of the environment with pathogenic properties. The results presented show evidence that molecular and cell-mediated immunological events in non-immune piglets have the initial stage of the specific progress of immune response against PEDv.

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Genetic Variant of SARS-CoV-2 Isolates in Indonesia: Spike Glycoprotein Gene

Journal of Pure and Applied Microbiology ◽

10.22207/jpam.14.spl1.35 ◽

2020 ◽

Vol 14 (suppl 1) ◽

pp. 971-978

Author(s):

Arif Nur Muhammad Ansori ◽

Viol Dhea Kharisma ◽

Sahal Sabilil Muttaqin ◽

Yulanda Antonius ◽

Arli Aditya Parikesit

Keyword(s):

Evolutionary Genetics ◽

Genetic Composition ◽

Spike Glycoprotein ◽

Glycoprotein Gene ◽

Antiviral Medication ◽

Surveillance Programs ◽

The Republic ◽

The Government ◽

Novel Coronavirus ◽

Gene Similarity

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), a novel coronavirus and the primary causative agent of coronavirus disease 2019 (COVID-19), first occurred in China and rapidly spread worldwide. The government of the Republic of Indonesia confirmed its first two cases of COVID-19 in March 2020. COVID-19 is a serious illness with no efficacious antiviral medication or approved vaccine currently available. Therefore, there is a need to investigate the genome of SARS-CoV-2. In this study, we characterized SARS-CoV-2 spike glycoprotein genes from Indonesia to investigate their genetic composition and variability. Overall, ten SARS-CoV-2 spike glycoprotein gene sequences retrieved from GenBank (National Center for Biotechnology Information, USA) and the GISAID EpiCoV database (Germany) were compared. We analyzed nucleotide variants and amino acid changes using Molecular Evolutionary Genetics Analysis (MEGA) X and analyzed gene similarity using the LALIGN web server. Interestingly, we revealed several specific mutation sites, however, there were no significant changes in the genetic composition of SARS-CoV-2 spike glycoprotein genes, when compared to the Wuhan-Hu-1 isolate from China. However, this is a preliminary study and we recommend that molecular epidemiology and surveillance programs against COVID-19 in Indonesia be improved.

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Clinical Trial of Glucose-Oral Rehydration Solution (ORS), Rice Dextrin-ORS, and Rice Flour-ORS for the Management of Children With Acute Diarrhea and Mild or Moderate Dehydration

PEDIATRICS ◽

10.1542/peds.95.2.191 ◽

1995 ◽

Vol 95 (2) ◽

pp. 191-197

Author(s):

Susana Molina ◽

Carolina Vettorazzi ◽

Janet M. Peerson ◽

Noel W. Solomons ◽

Kenneth H. Brown

Keyword(s):

Clinical Trial ◽

Treatment Group ◽

Acute Diarrhea ◽

Oral Rehydration Solution ◽

Rice Flour ◽

Watery Diarrhea ◽

Hydration Status ◽

Oral Rehydration ◽

Acute Watery Diarrhea ◽

Rehydration Solution

Objective. To assess the effects of glucose (G)-oral rehydration solution (ORS), rice dextrin (RD)-ORS, and rice flour (RF)-ORS on fluid intake, rapidity of rehydration, and stool output of children with acute diarrhea and mild or moderate dehydration. Methods. The study was a randomized, double-masked clinical trial. One hundred forty-six male infants, ages 3 to 36 months, were randomly assigned to one of three treatment groups. Clinical evaluations and fluid balances were conducted every 2 to 4 hours for 48 hours. Principal outcome variables were ORS consumption, recovery of hydration status, and fecal output. Results. The groups were similar at admission with regard to age, nutritional status, history of the current episode, and clinical status. There were no differences in ORS consumption by treatment group during any period of study. During the first 6-hour period, patients in group RF had less stool output (16 ± 14 g/kg/body weight) than those in group G (22 ± 20 g/kg) or RD (21 ± 19 g/kg; P < .05). After 12 hours of hospitalization, there were no differences by treatment group. Recovery of hydration status, changes in serum sodium and potassium, and duration of diarrhea in the hospital were similar in all three groups. Conclusion. There was a 24% to 27% reduction in stool output during the first 6 hours of treatment among children who received RF-ORS compared with those who received G-ORS or RD-ORS, but this effect did not persist after the first 12 hours of therapy. Because this difference was of small magnitude and limited duration, it has minor clinical importance. Thus, we conclude that the three solutions had similar efficacy for children with acute, watery diarrhea and mild or moderate dehydration.

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Discrimination of the SARS–CoV-2 strains using of coloured s-LASCA-imaging of GB-speckles, developed for the gene “S” nucleotide sequences

F1000Research ◽

10.12688/f1000research.53214.2 ◽

2021 ◽

Vol 10 ◽

pp. 503

Author(s):

Onega Ulianova ◽

Yury Saltykov ◽

Sergey Ulyanov ◽

Sergey Zaytsev ◽

Alexander Ulyanov ◽

...

Keyword(s):

Speckle Interferometry ◽

Molecular Target ◽

Nucleotide Sequences ◽

Considerable Improvement ◽

Spike Glycoprotein ◽

High Effectiveness ◽

Glycoprotein Gene ◽

Colour Image ◽

High Discrimination ◽

Pathogenic Viruses

Background: A recent bioinformatics technique involves changing nucleotide sequences into 2D speckles. This technique produces speckles called GB-speckles (Gene Based speckles). All classical strategies of speckle-optics, namely speckle-interferometry, subtraction of speckle-images as well as speckle-correlometry have been inferred for processing of GB-speckles. This indicates the considerable improvement in the present tools of bioinformatics. Methods: Colour s-LASCA imaging of virtual laser GB-speckles, a new method of high discrimination and typing of pathogenic viruses, has been developed. This method has been adapted to the detecting of natural mutations in nucleotide sequences, related to the spike glycoprotein (coding the gene «S») of SARS–CoV-2 gene as the molecular target. Results: The rate of the colouring images of virtual laser GB-speckles generated by s-LASCA can be described by the specific value of R. If the nucleotide sequences compared utilizing this approach the relevant images are completely identical, then the three components of the resulting colour image will be identical, and therefore the value of R will be equal to zero. However, if there are at least minimal differences in the matched nucleotide sequences, then the value of R will be positive. Conclusion: The high effectiveness of an application of the colour images of GB-speckles that were generated by s-LASCA- has been demonstrated for discrimination between different variants of the SARS–CoV-2 spike glycoprotein gene.

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The Incidence of Infection Among Contacts of Poliomyelitis Cases

PEDIATRICS ◽

10.1542/peds.17.1.36 ◽

1956 ◽

Vol 17 (1) ◽

pp. 36-36

Keyword(s):

Neutralizing Antibodies ◽

Virus Isolation ◽

Family Type ◽

Infection Rates ◽

Household Contacts ◽

Serological Examination ◽

The Family ◽

Blood Specimens ◽

Close Contacts

Employing tissue culture technic, the infection rates among close contacts of cases of poliomyelitis was studied during an epidemic. These studies were supplemented with determination of the presence of neutralizing antibodies against poliomyelitis virus in blood specimens. Twenty-four of fifty-four familial associates were found to be positive for the family type of virus; of those from whom no virus was isolated, 25 already had antibodies. Ninety-one per cent of family contacts were either excreting virus or possessed antibodies to the family type. The relation of symptoms to the isolation of virus and the ratio of inapparent to apparent infection among contacts was also studied. The infection rate for household contacts under 15 years of age was 100 per cent. The ratio of inapparent to apparent infection was approximately 3 to 1. The author states that, on the basis of virus isolation and serological examination, poliomyelitis may be considered as contagious as measles among susceptible individuals in a family situation.

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Effective screening of SARS-CoV-2 neutralizing antibodies in patient serum using lentivirus particles pseudotyped with SARS-CoV-2 spike glycoprotein

Scientific Reports ◽

10.1038/s41598-020-76135-w ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Ritesh Tandon ◽

Dipanwita Mitra ◽

Poonam Sharma ◽

Martin G. McCandless ◽

Stephen J. Stray ◽

...

Keyword(s):

Mammalian Cells ◽

Neutralizing Antibodies ◽

Transduction Efficiency ◽

Pseudotyped Virus ◽

Spike Glycoprotein ◽

Highly Pathogenic ◽

Mammalian Cell Lines ◽

Serological Studies ◽

Pathogenic Viruses ◽

Vesicular Stomatitis

Abstract Pseuodotyped particles have significant importance and use in virology as tools for studying the biology of highly pathogenic viruses in a lower biosafety environment. The biological, chemical, and serological studies of the recently emerged SARS-CoV-2 will be greatly aided by the development and optimization of a suitable pseudotyping system. Here, we pseudotyped the SARS-CoV-2 Spike glycoprotein (SPG) on a traditional retroviral (MMLV) as well as a third generation lentiviral (pLV) vector and tested the transduction efficiency in several mammalian cell lines expressing SARS-CoV-2 receptor hACE2. While MMLV pseudotyped the vesicular stomatitis virus G glycoprotein (VSV-G) efficiently, it could not pseudotype the full-length SPG. In contrast, pLV pseudotyped both glycoproteins efficiently; however, much higher titers of pLV-G particles were produced. Among all the tested mammalian cells, 293Ts expressing hACE2 were most efficiently transduced using the pLV-S system. The pLV-S particles were efficiently neutralized by diluted serum (>:640) from recently recovered COVID-19 patients who showed high SARS-CoV-2 specific IgM and IgG levels. In summary, pLV-S pseudotyped virus provides a valid screening tool for the presence of anti SARS-CoV-2 specific neutralizing antibodies in convalescent patient serum.

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Optimized Pseudotyping Conditions for the SARS-COV-2 Spike Glycoprotein

Journal of Virology ◽

10.1128/jvi.01062-20 ◽

2020 ◽

Vol 94 (21) ◽

Cited By ~ 8

Author(s):

Marc C. Johnson ◽

Terri D. Lyddon ◽

Reinier Suarez ◽

Braxton Salcedo ◽

Mary LePique ◽

...

Keyword(s):

Signal Peptide ◽

Neutralizing Antibodies ◽

Particle Production ◽

Surface Glycoprotein ◽

Spike Protein ◽

Target Cells ◽

Spike Glycoprotein ◽

Infectious Particle ◽

Pathogenic Virus ◽

Hiv 1

ABSTRACT The severe acute respiratory syndrome coronavirus 2 (SARS-COV-2) Spike glycoprotein is solely responsible for binding to the host cell receptor and facilitating fusion between the viral and host membranes. The ability to generate viral particles pseudotyped with SARS-COV-2 Spike is useful for many types of studies, such as characterization of neutralizing antibodies or development of fusion-inhibiting small molecules. Here, we characterized the use of a codon-optimized SARS-COV-2 Spike glycoprotein for the generation of pseudotyped HIV-1, murine leukemia virus (MLV), and vesicular stomatitis virus (VSV) particles. The full-length Spike protein functioned inefficiently with all three systems but was enhanced over 10-fold by deleting the last 19 amino acids of the cytoplasmic tail. Infection of 293FT target cells was possible only if the cells were engineered to stably express the human angiotensin-converting enzyme 2 (ACE2) receptor, but stably introducing an additional copy of this receptor did not further enhance susceptibility. Stable introduction of the Spike-activating protease TMPRSS2 further enhanced susceptibility to infection by 5- to 10-fold. Replacement of the signal peptide of the Spike protein with an optimal signal peptide did not enhance or reduce infectious particle production. However, modifications D614G and R682Q further enhanced infectious particle production. With all enhancing elements combined, the titer of pseudotyped HIV-1 particles reached almost 106 infectious particles/ml. Finally, HIV-1 particles pseudotyped with SARS-COV-2 Spike were successfully used to detect neutralizing antibodies in plasma from coronavirus disease 2019 (COVID-19) patients, but not in plasma from uninfected individuals. IMPORTANCE In work with pathogenic viruses, it is useful to have rapid quantitative tests for viral infectivity that can be performed without strict biocontainment restrictions. A common way of accomplishing this is to generate viral pseudoparticles that contain the surface glycoprotein from the pathogenic virus incorporated into a replication-defective viral particle that contains a sensitive reporter system. These pseudoparticles enter cells using the glycoprotein from the pathogenic virus, leading to a readout for infection. Conditions that block entry of the pathogenic virus, such as neutralizing antibodies, will also block entry of the viral pseudoparticles. However, viral glycoproteins often are not readily suited for generating pseudoparticles. Here, we describe a series of modifications that result in the production of relatively high-titer SARS-COV-2 pseudoparticles that are suitable for the detection of neutralizing antibodies from COVID-19 patients.

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Epitope mapping and cellular localization of swine acute diarrhea syndrome coronavirus nucleocapsid protein using a novel monoclonal antibody

Virus Research ◽

10.1016/j.virusres.2019.197752 ◽

2019 ◽

Vol 273 ◽

pp. 197752

Author(s):

Yuru Han ◽

Jiyu Zhang ◽

Hongyan Shi ◽

Ling Zhou ◽

Jianfei Chen ◽

...

Keyword(s):

Monoclonal Antibody ◽

Acute Diarrhea ◽

Nucleocapsid Protein ◽

Cellular Localization ◽

Epitope Mapping ◽

Diarrhea Syndrome

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1491. Active Norovirus Surveillance in Children Under 5 Years with Diarrhea after Rotavirus Vaccine Introduction in Argentina (2017–2019)

Open Forum Infectious Diseases ◽

10.1093/ofid/ofz360.1355 ◽

2019 ◽

Vol 6 (Supplement_2) ◽

pp. S543-S543

Author(s):

Angela Gentile ◽

Juan Stupka ◽

Juan I Degiuseppe ◽

María del Valle Juárez ◽

Maria Florencia Lucion ◽

...

Keyword(s):

Acute Diarrhea ◽

Epidemiological Data ◽

Cross Sectional Study ◽

Epidemiological Surveillance ◽

Watery Diarrhea ◽

Rotavirus Vaccine ◽

Cross Sectional ◽

Vaccine Introduction ◽

Rotavirus Vaccination ◽

Children Under 5

Abstract Background Acute diarrhea is one of the leading causes of infant morbidity and mortality. Argentina introduced massive rotavirus vaccination in 2015. In several countries, this introduction has changed the distribution of enteropathogens. The decrease in the prevalence of rotavirus has been described at the expense of an increase in Norovirus (NoV) activity worldwide. The aim of this study was to analyze the role of NoV in acute diarrhea cases in outpatient children under 5 years of age and their epidemiological profile. Methods A prospective and cross-sectional study in <5 years outpatients attended for acute diarrhea in Children’s Hospital “Dr. Ricardo Gutiérrez” in Buenos Aires, Argentina, between July 2017 and March 2019 was conducted. Active epidemiological surveillance was performed with a specific case reporting form. Stool samples were tested for NoV (RT-qPCR). Clinical and epidemiological data were recorded. Results A total of 252 patients were enrolled and 235 stools samples were tested. Median of age was 22.3 months (IQR: 11–30), 58.7% were male. The most frequent symptoms were fever and vomiting in 63.1% and 53%, respectively; 52% had watery diarrhea, 45.2% had moderate diarrhea according to Vesikari Scale, 95.6% were normohydrated and 22% had a household member with diarrhea. There were no immunocompromised children. A 72% had received rotavirus vaccine, 86% of them with full scheme. From samples tested, 27% (n = 63) were NoV positive. NoV was found throughout the year and the frequency of detection was higher in January and June (summer and winter in Argentina). Regarding genetic diversity the most frequent genogroup was GII (65%; 41/63) and genotype GII.P16-GII.4 Sydney (48%; 20/41). Bacterial co-infection was observed in 35%. Compared with negative cases, NoV were younger (18 vs. 20 months; P < 0.001) and were associated with higher prevalence of rotavirus vaccination (88% vs. 66%; P = 0.001). No statistically difference was found regarding to gender, clinical outcome and severity. Conclusion NoV was detected at high frequency (27%) in children presenting moderate acute diarrhea, mainly in those who received rotavirus vaccine. Regarding sporadic acute diarrhea cases in children, it is important to consider NoV as a frequent etiological agent. Disclosures All authors: No reported disclosures.

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