The Use of Convalescent Sera in Immune-Electron Microscopy to Detect Non-Suspected/New Viral Agents

Llombart‐Bosch, Antonio; Tittarelli, Claudia Marcela; Cerioli, M.

doi:10.3390/v7052683

Cited by 16 publications

(18 citation statements)

References 32 publications

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“…5B). After PEDV infection in IPEC-J2 cells, viral particles assumed to be PEDV due to their mostly round shape (Lavazza et al, 2015) and diameter ranging from 95 to 190 nm (Lee, 2015) were found in the cytosol of the cells (Fig. 5B).…”

Section: Pedv Is a Potential Target Of Autophagymentioning

confidence: 99%

Rapamycin-induced autophagy restricts porcine epidemic diarrhea virus infectivity in porcine intestinal epithelial cells

Kim

et al. 2017

Antiviral Research

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Porcine epidemic diarrhea virus (PEDV) invades porcine intestinal epithelial cells (IECs) and causes diarrhea and dehydration in pigs. In the present study, we showed a suppression of PEDV infection in porcine jejunum intestinal epithelial cells (IPEC-J2) by an increase in autophagy. Autophagy was activated by rapamycin at a dose that does not affect cell viability and tight junction permeability. The induction of autophagy was examined by LC3I/LC3II conversion. To confirm the autophagic-flux (entire autophagy pathway), autophagolysosomes were examined by an immunofluorescence assay. Pre-treatment with rapamycin significantly restricted not only a 1 h infection but also a longer infection (24 h) with PEDV, while this effect disappeared when autophagy was blocked. Co-localization of PEDV and autophagosomes suggests that PEDV could be a target of autophagy. Moreover, alleviation of PEDV-induced cell death in IPEC-J2 cells pretreated with rapamycin demonstrates a protective effect of rapamycin against PEDV-induced epithelial cell death. Collectively, the present study suggests an early prevention against PEDV infection in IPEC-J2 cells via autophagy that might be an effective strategy for the restriction of PEDV, and opens up the possibility of the use of rapamycin in vivo as an effective prophylactic and prevention treatment.

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Section: Pedv Is a Potential Target Of Autophagymentioning

confidence: 99%

Rapamycin-induced autophagy restricts porcine epidemic diarrhea virus infectivity in porcine intestinal epithelial cells

Kim

et al. 2017

Antiviral Research

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“…Immunoelectron microscopy (IEM) is utilized when the number of viral particles in a sample is very low, when virions are pleomorphic and difficult to identify because they do not have a typical viral morphology or when the samples are "dirty" because the aggregated complexes are more easily observed [203]. It allows identification of the virus for specific antigen-antibody reaction and such identification is achieved by its morphology.…”

Section: Immunoelectron Microscopy Techniquementioning

confidence: 99%

“…Several variations of the method such as immune clumping or direct immunoelectron microscopy (DIEM) [206,207] or immune aggregation electron microscopy (IAEM) [203], solid phase immune electron microscopy (SPIEM) [199] and decoration [208] have been used. Hyperimmune sera, monoclonal antibodies or convalescent sera can be used in performing the technique [193,203].…”

Section: Immunoelectron Microscopy Techniquementioning

confidence: 99%

“…Hyperimmune sera, monoclonal antibodies or convalescent sera can be used in performing the technique [193,203]. The SPIEM has been utilized to detect most of the viruses that cause gastroenteritis such as bovine rotavirus, swine, equine, canine, bovine coronavirus, swine, canine parvovirus and BVDV.…”

Section: Immunoelectron Microscopy Techniquementioning

confidence: 99%

“…IAEM was used to detect porcine rotavirus (PoRV), porcine torovirus (PoToV) and porcine epidemic diarrhea coronavirus (PEDV) in pigs with enteritis utilizing convalescent sera [203].…”

Section: Immunoelectron Microscopy Techniquementioning

confidence: 99%

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Application of Transmission Electron Microscopy Techniques in the Veterinary Diagnosis of Viral Gastroenteritis in Livestock Animals

Catroxo¹,

Martins²

2018

Stomach Disorders

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Gastroenteritis caused by viruses is considered to be one of the most important diseases in livestock, being the main cause of morbidity and mortality in young animals, culminating in serious economic losses due to costs with prophylaxis and treatment, increased susceptibility of animals to secondary infections, developmental delay and death. Stressful factors may support the onset of illness. Several viral agents can cause gastroenteritis in various animal species. Rotaviruses are considered the main cause of enteric infections in various animals, including humans constituting important zoonosis. Due to genetic diversity and their ability to cross the species barrier, the coronaviruses infect many species. In cattle, they cause "Winter Dysentery" in adult animals and "Neonatal Diarrhea" in newborn calves. In swine, they are responsible for "Transmissible Gastroenteritis" and "Swine Epidemic Diarrhea." Equines infected with coronavirus also develop severe gastroenteritis. Bovine viral diarrhea (BVD) caused by a flavivirus of the genus Pestivirus is related to digestive and reproductive disorders, affecting any productive sector, are it cut, milk or confinement. Transmission electron microscopy is an indispensable tool in the diagnosis of viral gastroenteric infectious diseases. Negative staining is a simple, fast and efficient technique, being ideal for the detection of gastroenteric viruses, being easily visualized. The immunoelectron microscopy (IEM) technique allows increasing the sensitivity of virus detection where low concentrations of virus are aggregated so that they may be more easily seen. The immunolabeling with colloidal gold technique utilizes specific antibodies tagged with particles of colloidal gold to label the antigen antibody reaction. Embedding resin technique allows obtaining information on the virus-cell interaction. The different transmission electron microscopy modalities promotes a fast and accurate diagnosis of the different gastroenteric viral agents, allowing prophylactic measures of control and prevention in the creations to be promptly instituted, avoiding animal losses and disastrous economic losses, and collaborating with the National Porcine and Bovine Agribusiness.

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Virus detection by transmission electron microscopy: Still useful for diagnosis and a plus for biosafety

Roingeard

Raynal

Eymieux

et al. 2018

Reviews in Medical Virology

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Summary Transmission electron microscopy (TEM) is the only imaging technique allowing the direct visualization of viruses, due to its nanometer‐scale resolution. Between the 1960s and 1990s, TEM contributed to the discovery of many types of viruses and served as a diagnostic tool for identifying viruses directly in biological samples, either in suspension or in sections of tissues or mammalian cells grown in vitro in contact with clinical samples. The diagnosis of viral infections improved considerably during the 1990s, with the advent of highly sensitive techniques, such as enzyme‐linked immunosorbent assay (ELISA) and PCR, rendering TEM obsolete for this purpose. However, the last 20 years have demonstrated the utility of this technique in particular situations, due to its “catch‐all” nature, making diagnosis possible through visualization of the virus, without the need of prior assumptions about the infectious agent sought. Thus, in several major outbreaks in which molecular techniques failed to identify the infectious agent, TEM provided the answer. TEM is also still occasionally used in routine diagnosis to characterize infections not diagnosed by molecular assays. It is also used to check the microbiological safety of biological products. Many biopharmaceuticals are produced in animal cells that might contain little‐known, difficult‐to‐detect viruses. In this context, the “catch‐all” properties of TEM make it possible to document the presence of viruses or virus‐like particles in these products.

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The Use of Convalescent Sera in Immune-Electron Microscopy to Detect Non-Suspected/New Viral Agents

Cited by 16 publications

References 32 publications

Rapamycin-induced autophagy restricts porcine epidemic diarrhea virus infectivity in porcine intestinal epithelial cells

Rapamycin-induced autophagy restricts porcine epidemic diarrhea virus infectivity in porcine intestinal epithelial cells

Application of Transmission Electron Microscopy Techniques in the Veterinary Diagnosis of Viral Gastroenteritis in Livestock Animals

Virus detection by transmission electron microscopy: Still useful for diagnosis and a plus for biosafety

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