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Solid phase immune electron microscopy for diagnosis of transmissible gastroenteritis in pigs
A serological trapping technique is described for detecting transmissible gastroenteritis (tge) virus in faeces. The technique involves the coating of electron microscope grids with protein A and specific tge virus antiserum. Optimal conditions for performing this solid phase immune electron microsc...
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Published by Elsevier Ltd.
1988
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7131200/ https://www.ncbi.nlm.nih.gov/pubmed/2841730 http://dx.doi.org/10.1016/S0034-5288(18)30859-2 |
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author | VAN NIEUWSTADT, A.P. CORNELISSEN, J.B.W.J. VREESWIJK, J. |
author_facet | VAN NIEUWSTADT, A.P. CORNELISSEN, J.B.W.J. VREESWIJK, J. |
author_sort | VAN NIEUWSTADT, A.P. |
collection | PubMed |
description | A serological trapping technique is described for detecting transmissible gastroenteritis (tge) virus in faeces. The technique involves the coating of electron microscope grids with protein A and specific tge virus antiserum. Optimal conditions for performing this solid phase immune electron microscopy technique were a concentration 250 µg ml(−1) of protein A; 1:100 diluted rabbit anti-tge virus hyperimmune serum for coating the grids and overnight incubation with virus samples. The possibility of detecting coronavirus in crude faeces was highly improved with solid phase immune electron microscopy, compared with conventional negative staining electron microscopy, by specific trapping of virus and prevention of adsorption of contaminants. The sensitivity of the method was evaluated by coded investigation of a dilution series of stock virus mixed with different pools of faeces. The improvement of virus detection in faeces by solid phase immune electron microscopy, compared with standard electron microscopy was at least 100-fold. Faecal shedding of coronavirus by pigs infected with virulent and attenuated strains of tge virus was studied. Virus detection in faeces by a standard electron microscopy technique was not practical, since the virus was obscured by a large quantity of debris in the faeces. By using solid phase immune electron microscopy, however, the aspect of the specimens on the grids improved so much that, in addition to more common immunofluorescence, the technique might be useful as a diagnostic test for tge. Virus was detected in daily faecal samples from one or two days after experimental infection with virulent tge virus, until death in five out of eight animals. Unlike immunofluorescence, solid phase immune electron microscopy may be used for diagnosis in living animals. |
format | Online Article Text |
id | pubmed-7131200 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 1988 |
publisher | Published by Elsevier Ltd. |
record_format | MEDLINE/PubMed |
spelling | pubmed-71312002020-04-08 Solid phase immune electron microscopy for diagnosis of transmissible gastroenteritis in pigs VAN NIEUWSTADT, A.P. CORNELISSEN, J.B.W.J. VREESWIJK, J. Res Vet Sci Article A serological trapping technique is described for detecting transmissible gastroenteritis (tge) virus in faeces. The technique involves the coating of electron microscope grids with protein A and specific tge virus antiserum. Optimal conditions for performing this solid phase immune electron microscopy technique were a concentration 250 µg ml(−1) of protein A; 1:100 diluted rabbit anti-tge virus hyperimmune serum for coating the grids and overnight incubation with virus samples. The possibility of detecting coronavirus in crude faeces was highly improved with solid phase immune electron microscopy, compared with conventional negative staining electron microscopy, by specific trapping of virus and prevention of adsorption of contaminants. The sensitivity of the method was evaluated by coded investigation of a dilution series of stock virus mixed with different pools of faeces. The improvement of virus detection in faeces by solid phase immune electron microscopy, compared with standard electron microscopy was at least 100-fold. Faecal shedding of coronavirus by pigs infected with virulent and attenuated strains of tge virus was studied. Virus detection in faeces by a standard electron microscopy technique was not practical, since the virus was obscured by a large quantity of debris in the faeces. By using solid phase immune electron microscopy, however, the aspect of the specimens on the grids improved so much that, in addition to more common immunofluorescence, the technique might be useful as a diagnostic test for tge. Virus was detected in daily faecal samples from one or two days after experimental infection with virulent tge virus, until death in five out of eight animals. Unlike immunofluorescence, solid phase immune electron microscopy may be used for diagnosis in living animals. Published by Elsevier Ltd. 1988-05 2018-08-23 /pmc/articles/PMC7131200/ /pubmed/2841730 http://dx.doi.org/10.1016/S0034-5288(18)30859-2 Text en Copyright © 1988 Published by Elsevier Ltd. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active. |
spellingShingle | Article VAN NIEUWSTADT, A.P. CORNELISSEN, J.B.W.J. VREESWIJK, J. Solid phase immune electron microscopy for diagnosis of transmissible gastroenteritis in pigs |
title | Solid phase immune electron microscopy for diagnosis of transmissible gastroenteritis in pigs |
title_full | Solid phase immune electron microscopy for diagnosis of transmissible gastroenteritis in pigs |
title_fullStr | Solid phase immune electron microscopy for diagnosis of transmissible gastroenteritis in pigs |
title_full_unstemmed | Solid phase immune electron microscopy for diagnosis of transmissible gastroenteritis in pigs |
title_short | Solid phase immune electron microscopy for diagnosis of transmissible gastroenteritis in pigs |
title_sort | solid phase immune electron microscopy for diagnosis of transmissible gastroenteritis in pigs |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7131200/ https://www.ncbi.nlm.nih.gov/pubmed/2841730 http://dx.doi.org/10.1016/S0034-5288(18)30859-2 |
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