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Isolation of peste des petits ruminants virus using primary goat kidney cell culture from kidneys obtained at slaughter

BACKGROUND: Traditionally isolation of peste des petits ruminant virus (PPRV) is performed in Vero cells that takes several blind passages before observing typical cytopathic effects (CPEs). As an alternate, researchers have been using lamb kidney (LK) cells but day‐old lambs are difficult to obtain...

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Autores principales: Begum, Shahana, Nooruzzaman, Mohammed, Hasnat, Azmary, Parvin, Mst. Murshida, Parvin, Rokshana, Islam, Mohammad Rafiqul, Chowdhury, Emdadul Haque
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8136951/
https://www.ncbi.nlm.nih.gov/pubmed/33326709
http://dx.doi.org/10.1002/vms3.413
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author Begum, Shahana
Nooruzzaman, Mohammed
Hasnat, Azmary
Parvin, Mst. Murshida
Parvin, Rokshana
Islam, Mohammad Rafiqul
Chowdhury, Emdadul Haque
author_facet Begum, Shahana
Nooruzzaman, Mohammed
Hasnat, Azmary
Parvin, Mst. Murshida
Parvin, Rokshana
Islam, Mohammad Rafiqul
Chowdhury, Emdadul Haque
author_sort Begum, Shahana
collection PubMed
description BACKGROUND: Traditionally isolation of peste des petits ruminant virus (PPRV) is performed in Vero cells that takes several blind passages before observing typical cytopathic effects (CPEs). As an alternate, researchers have been using lamb kidney (LK) cells but day‐old lambs are difficult to obtain and requires animal sacrifice. OBJECTIVE: We established a primary goat kidney (GK) cell culture from the kidneys obtained at slaughter. METHODS: The kidney of Black Bengal goats were collected from slaughter house and processed to make single cell suspension. The cells were resuspended in appropriate culture medium and maintained under optimum culture condition. RESULTS: The 80% confluent monolayer of GK cells was obtained after 15–20 days post seeding. Upon infection with a field isolate of PPRV, the well‐developed CPEs characterized by cell rounding, vacuolation in the cytoplasm and fusion of cells were observed after 48 hr post infection. Virus quantification in the culture supernatant revealed more viral RNA in GK cells than LK cells. The multicycle growth analysis of PPRV showed a steady increase in the virus loads in the culture supernatant of infected GK cells, suggesting an adaptation of the PPRV in GK cells. CONCLUSIONS: The findings suggest that primary GK cells can be successfully prepared from the mature kidney cortical tissues and can be used for the isolation of PPRV. This system could reduce the unnecessary sacrifice of lambs or kids. Since kidneys of slaughtered goats are available throughout the year, using this protocol primary cell culture from mature goat kidney can provide primary cells to the laboratory throughout the year.
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spelling pubmed-81369512021-05-24 Isolation of peste des petits ruminants virus using primary goat kidney cell culture from kidneys obtained at slaughter Begum, Shahana Nooruzzaman, Mohammed Hasnat, Azmary Parvin, Mst. Murshida Parvin, Rokshana Islam, Mohammad Rafiqul Chowdhury, Emdadul Haque Vet Med Sci Original Articles BACKGROUND: Traditionally isolation of peste des petits ruminant virus (PPRV) is performed in Vero cells that takes several blind passages before observing typical cytopathic effects (CPEs). As an alternate, researchers have been using lamb kidney (LK) cells but day‐old lambs are difficult to obtain and requires animal sacrifice. OBJECTIVE: We established a primary goat kidney (GK) cell culture from the kidneys obtained at slaughter. METHODS: The kidney of Black Bengal goats were collected from slaughter house and processed to make single cell suspension. The cells were resuspended in appropriate culture medium and maintained under optimum culture condition. RESULTS: The 80% confluent monolayer of GK cells was obtained after 15–20 days post seeding. Upon infection with a field isolate of PPRV, the well‐developed CPEs characterized by cell rounding, vacuolation in the cytoplasm and fusion of cells were observed after 48 hr post infection. Virus quantification in the culture supernatant revealed more viral RNA in GK cells than LK cells. The multicycle growth analysis of PPRV showed a steady increase in the virus loads in the culture supernatant of infected GK cells, suggesting an adaptation of the PPRV in GK cells. CONCLUSIONS: The findings suggest that primary GK cells can be successfully prepared from the mature kidney cortical tissues and can be used for the isolation of PPRV. This system could reduce the unnecessary sacrifice of lambs or kids. Since kidneys of slaughtered goats are available throughout the year, using this protocol primary cell culture from mature goat kidney can provide primary cells to the laboratory throughout the year. John Wiley and Sons Inc. 2020-12-16 /pmc/articles/PMC8136951/ /pubmed/33326709 http://dx.doi.org/10.1002/vms3.413 Text en © 2020 The Authors Veterinary Medicine and Science Published by John Wiley & Sons Ltd https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Articles
Begum, Shahana
Nooruzzaman, Mohammed
Hasnat, Azmary
Parvin, Mst. Murshida
Parvin, Rokshana
Islam, Mohammad Rafiqul
Chowdhury, Emdadul Haque
Isolation of peste des petits ruminants virus using primary goat kidney cell culture from kidneys obtained at slaughter
title Isolation of peste des petits ruminants virus using primary goat kidney cell culture from kidneys obtained at slaughter
title_full Isolation of peste des petits ruminants virus using primary goat kidney cell culture from kidneys obtained at slaughter
title_fullStr Isolation of peste des petits ruminants virus using primary goat kidney cell culture from kidneys obtained at slaughter
title_full_unstemmed Isolation of peste des petits ruminants virus using primary goat kidney cell culture from kidneys obtained at slaughter
title_short Isolation of peste des petits ruminants virus using primary goat kidney cell culture from kidneys obtained at slaughter
title_sort isolation of peste des petits ruminants virus using primary goat kidney cell culture from kidneys obtained at slaughter
topic Original Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8136951/
https://www.ncbi.nlm.nih.gov/pubmed/33326709
http://dx.doi.org/10.1002/vms3.413
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