The role of electron microscopy in the rapid diagnosis of viral infections — review

Schramlová, J; Arientová, Simona; Hulínská, D

doi:10.1007/s12223-010-0015-8

Cited by 37 publications

(36 citation statements)

References 43 publications

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“…rtPCR has become the mainstream diagnostic approach for many viral pathogens but EM and EIA remain as routine diagnostic tests for EV in many laboratories. The comparison of rtPCR was restricted to EM as EM is the routine diagnostic test currently used for sporadic gastroenteritis screening at Provincial Laboratory for Public Health, virology reference laboratory for Alberta, as well as other similar laboratories in Canada and internationally [Schramlová et al, ; Higgins et al, ]. A three‐way evaluation comparing EM, EIA, and molecular methods was not carried out in this study because of resource limitations.…”

Section: Discussionmentioning

confidence: 99%

Enhanced enteric virus detection in sporadic gastroenteritis using a multi-target real-time PCR panel: A one-year study

2013

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Viral gastroenteritis causes significant mortality and morbidity worldwide. Identifying the etiology of viral gastroenteritis is a challenge as most enteric viruses (EVs) are non-culturable. This study is to develop an EV testing panel using real-time PCR (EVPrtPCR) to simultaneously detect rotavirus, norovirus, sapovirus, astrovirus, and enteric adenovirus in stool samples. EVPrtPCR using universal amplification conditions was run in a single instrument run. EVPrtPCR was used to test 2,486 sporadic gastroenteritis samples submitted for EV testing using electron microscopy (EM) between July 2008 and July 2009. Retesting spiked negative stool samples and Salmon DNA as internal control were used to evaluate inhibition. EVPrtPCR detected viruses in significantly more samples: 748 (34%) as compared to 94 (3.8%) by EM. EM did not detect any norovirus, sapovirus, and mixed infection, and detected only 39% of rotavirus and 38.2% of enteric adenovirus positive samples. Four samples that tested positive for rotavirus and two for adenovirus and for small-round-structured viruses by EM were negative by EVPrtPCR. Norovirus was the most common virus detected (17.6%) with 92.4% as genogroup II, followed by rotavirus (6.8%), sapovirus (4.2%), astrovirus (2.0%), and enteric adenovirus (1.4%) with 9% samples positive for mixed infection. Overall, EV identification followed a U-shaped age distribution; positive samples were more common in children ≤5 years old and adults >60 years old. Norovirus, sapovirus and astrovirus showed winter predominance and rotavirus peaked in the spring. No inhibition was observed. Molecular technology significantly enhanced the identification of EV causing sporadic gastroenteritis in Alberta.

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Section: Discussionmentioning

confidence: 99%

Enhanced enteric virus detection in sporadic gastroenteritis using a multi-target real-time PCR panel: A one-year study

2013

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“…TEM images of presumptive naked Picornaviridae (enterovirus)‐like particles and (f) a known coxsackievirus , present in the Umgeni River at sampling sites U1, U2, and U3 during spring and summer tested. Images captured at 500 000–600 000× magnification.…”

Section: Resultsmentioning

confidence: 99%

“…TEM images of presumptive enveloped VLPs (a, b,c) Coronaviridae VLPs, (d) known coronavirus , (e, f, g) presumptive Orthomyxoviridae VLPs, (h) known influenzavirus , present in the Umgeni River at the different sampling sites during all seasons tested. Images captured at 400 000–600 000× magnification.…”

Section: Resultsmentioning

confidence: 99%

Detecting Virus‐Like Particles from the Umgeni River, South Africa

Ganesh

Lin

Singh

2013

CLEAN Soil Air Water

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It is important to consider viruses in water quality because of their incidence as causal agents for diarrhoeal disease, and due to their characteristics, which allow them to survive in changing environmental conditions indefinitely. This study assessed the viral quality of the Umgeni River in South Africa seasonally. A two‐step tangential flow filtration process was setup to remove the bacteria and to concentrate the virus populations from large volume water samples. The concentrated water samples contained up to 659 and 550 pfu/mL of somatic and F‐RNA coliphages, respectively. Several virus families including Adenoviridae, Herpesviridae, Orthomyxoviridae, Picornaviridae, Poxviridae and Reoviridae were found in the river based on the morphologies examined under transmission electron microscopy. All concentrated water samples produced substantial cytopathic effects on the Vero, HEK 293, Hela and A549 cell lines. These results indicate the potential of viruses in the water samples especially from the lower catchment areas of the Umgeni River to infect human hosts throughout the year. The present study highlights the importance of routine environmental surveillance of human enteric viruses in water sources. This can contribute to a better understanding of the actual burden of disease on those who might be using the water directly without treatment.

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“…Transmission electron microscopy is a perfectly adequate tool to investigate viral agents during outbreaks of gastroenteritis [191]. It is used when it is necessary to apply a fast and reliable diagnostic method to contain the infection and to quickly minimize the animal losses and consequently the economic damages that cause to the rational economic exploitation of the production animals, as much by the decrease of the productivity as in terms of treatment costs.…”

Section: Transmission Electron Microscopymentioning

confidence: 99%

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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The role of electron microscopy in the rapid diagnosis of viral infections — review

Cited by 37 publications

References 43 publications

Enhanced enteric virus detection in sporadic gastroenteritis using a multi-target real-time PCR panel: A one-year study

Enhanced enteric virus detection in sporadic gastroenteritis using a multi-target real-time PCR panel: A one-year study

Detecting Virus‐Like Particles from the Umgeni River, South Africa

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

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