Comparative Susceptibility of Madin–Darby Canine Kidney (MDCK) Derived Cell Lines for Isolation of Swine Origin Influenza A Viruses from Different Clinical Specimens
Madin–Darby canine kidney (MDCK) cells are commonly used for the isolation of mammalian influenza A viruses. The goal of this study was to compare the sensitivity and suitability of the original MDCK cell line in comparison with MDCK-derived cell lines, MDCK.2, MDCK SIAT-1 and MDCK-London for isolat...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2021-11-01
|
| Series: | Viruses |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1999-4915/13/12/2346 |
| _version_ | 1797500008707129344 |
|---|---|
| author | Matthew Suderman Mariko Moniwa Tamiru N. Alkie Davor Ojkic Andre Broes Neil Pople Yohannes Berhane |
| author_facet | Matthew Suderman Mariko Moniwa Tamiru N. Alkie Davor Ojkic Andre Broes Neil Pople Yohannes Berhane |
| author_sort | Matthew Suderman |
| collection | DOAJ |
| description | Madin–Darby canine kidney (MDCK) cells are commonly used for the isolation of mammalian influenza A viruses. The goal of this study was to compare the sensitivity and suitability of the original MDCK cell line in comparison with MDCK-derived cell lines, MDCK.2, MDCK SIAT-1 and MDCK-London for isolation of swine-origin influenza A viruses (IAV-S) from clinical specimens. One-hundred thirty clinical specimens collected from pigs in the form of nasal swabs, lung tissue and oral fluids that were positive by PCR for the presence of IAV-S RNA were inoculated in the cell cultures listed above. MDCK-SIAT1 cells yielded the highest proportion of positive IAV-S isolations from all specimen types. For nasal swabs, 58.62% of the specimens were IAV-S positive in MDCK-SIAT1 cells, followed by MDCK-London (36.21%), and conventional MDCK and MDCK.2 cells (27.5%). For lung specimens, 59.38% were IAV-S positive in MDCK-SIAT1 cells, followed by MDCK-London (40.63%), and conventional MDCK and MDCK.2 cells (18.75–31.25%). Oral fluids yielded the lowest number of positive virus isolation results, but MDCK-SIAT1 cells were still had the highest rate (35%) of IAV-S isolation, whereas the isolation rate in other cells ranged from 5–7.5%. Samples with lower IAV-S PCR cycle threshold (Ct) values were more suitable for culturing and isolation. The isolated IAV-S represented H1N1-β, H1N2-α, H1N1pdm and H3N2 cluster IV and cluster IVB viruses. The result of the current study demonstrated the importance of using the most appropriate MDCK cells when isolating IAV-S from clinical samples. |
| first_indexed | 2024-03-10T03:55:40Z |
| format | Article |
| id | doaj.art-f71611909b02442b86bc619284ce71ec |
| institution | Directory Open Access Journal |
| issn | 1999-4915 |
| language | English |
| last_indexed | 2024-03-10T03:55:40Z |
| publishDate | 2021-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Viruses |
| spelling | doaj.art-f71611909b02442b86bc619284ce71ec2023-11-23T10:56:35ZengMDPI AGViruses1999-49152021-11-011312234610.3390/v13122346Comparative Susceptibility of Madin–Darby Canine Kidney (MDCK) Derived Cell Lines for Isolation of Swine Origin Influenza A Viruses from Different Clinical SpecimensMatthew Suderman0Mariko Moniwa1Tamiru N. Alkie2Davor Ojkic3Andre Broes4Neil Pople5Yohannes Berhane6National Centre for Foreign Animal Disease, Canadian Food Inspection Agency, Winnipeg, MB R3E 3R2, CanadaNational Centre for Foreign Animal Disease, Canadian Food Inspection Agency, Winnipeg, MB R3E 3R2, CanadaNational Centre for Foreign Animal Disease, Canadian Food Inspection Agency, Winnipeg, MB R3E 3R2, CanadaAnimal Health Laboratory, University of Guelph, Guelph, ON N1G 2W1, CanadaBiovet Inc., Saint-Hyacinthe, QC J2S 8W2, CanadaVeterinary Diagnostic Services, Manitoba Agriculture and Resource Development, 545 University Crescent, Winnipeg, MB R3T 5S6, CanadaNational Centre for Foreign Animal Disease, Canadian Food Inspection Agency, Winnipeg, MB R3E 3R2, CanadaMadin–Darby canine kidney (MDCK) cells are commonly used for the isolation of mammalian influenza A viruses. The goal of this study was to compare the sensitivity and suitability of the original MDCK cell line in comparison with MDCK-derived cell lines, MDCK.2, MDCK SIAT-1 and MDCK-London for isolation of swine-origin influenza A viruses (IAV-S) from clinical specimens. One-hundred thirty clinical specimens collected from pigs in the form of nasal swabs, lung tissue and oral fluids that were positive by PCR for the presence of IAV-S RNA were inoculated in the cell cultures listed above. MDCK-SIAT1 cells yielded the highest proportion of positive IAV-S isolations from all specimen types. For nasal swabs, 58.62% of the specimens were IAV-S positive in MDCK-SIAT1 cells, followed by MDCK-London (36.21%), and conventional MDCK and MDCK.2 cells (27.5%). For lung specimens, 59.38% were IAV-S positive in MDCK-SIAT1 cells, followed by MDCK-London (40.63%), and conventional MDCK and MDCK.2 cells (18.75–31.25%). Oral fluids yielded the lowest number of positive virus isolation results, but MDCK-SIAT1 cells were still had the highest rate (35%) of IAV-S isolation, whereas the isolation rate in other cells ranged from 5–7.5%. Samples with lower IAV-S PCR cycle threshold (Ct) values were more suitable for culturing and isolation. The isolated IAV-S represented H1N1-β, H1N2-α, H1N1pdm and H3N2 cluster IV and cluster IVB viruses. The result of the current study demonstrated the importance of using the most appropriate MDCK cells when isolating IAV-S from clinical samples.https://www.mdpi.com/1999-4915/13/12/2346Madin–Darby canine kidney (MDCK) cellsvirus isolationswine influenzanasal swabsoral fluids |
| spellingShingle | Matthew Suderman Mariko Moniwa Tamiru N. Alkie Davor Ojkic Andre Broes Neil Pople Yohannes Berhane Comparative Susceptibility of Madin–Darby Canine Kidney (MDCK) Derived Cell Lines for Isolation of Swine Origin Influenza A Viruses from Different Clinical Specimens Viruses Madin–Darby canine kidney (MDCK) cells virus isolation swine influenza nasal swabs oral fluids |
| title | Comparative Susceptibility of Madin–Darby Canine Kidney (MDCK) Derived Cell Lines for Isolation of Swine Origin Influenza A Viruses from Different Clinical Specimens |
| title_full | Comparative Susceptibility of Madin–Darby Canine Kidney (MDCK) Derived Cell Lines for Isolation of Swine Origin Influenza A Viruses from Different Clinical Specimens |
| title_fullStr | Comparative Susceptibility of Madin–Darby Canine Kidney (MDCK) Derived Cell Lines for Isolation of Swine Origin Influenza A Viruses from Different Clinical Specimens |
| title_full_unstemmed | Comparative Susceptibility of Madin–Darby Canine Kidney (MDCK) Derived Cell Lines for Isolation of Swine Origin Influenza A Viruses from Different Clinical Specimens |
| title_short | Comparative Susceptibility of Madin–Darby Canine Kidney (MDCK) Derived Cell Lines for Isolation of Swine Origin Influenza A Viruses from Different Clinical Specimens |
| title_sort | comparative susceptibility of madin darby canine kidney mdck derived cell lines for isolation of swine origin influenza a viruses from different clinical specimens |
| topic | Madin–Darby canine kidney (MDCK) cells virus isolation swine influenza nasal swabs oral fluids |
| url | https://www.mdpi.com/1999-4915/13/12/2346 |
| work_keys_str_mv | AT matthewsuderman comparativesusceptibilityofmadindarbycaninekidneymdckderivedcelllinesforisolationofswineorigininfluenzaavirusesfromdifferentclinicalspecimens AT marikomoniwa comparativesusceptibilityofmadindarbycaninekidneymdckderivedcelllinesforisolationofswineorigininfluenzaavirusesfromdifferentclinicalspecimens AT tamirunalkie comparativesusceptibilityofmadindarbycaninekidneymdckderivedcelllinesforisolationofswineorigininfluenzaavirusesfromdifferentclinicalspecimens AT davorojkic comparativesusceptibilityofmadindarbycaninekidneymdckderivedcelllinesforisolationofswineorigininfluenzaavirusesfromdifferentclinicalspecimens AT andrebroes comparativesusceptibilityofmadindarbycaninekidneymdckderivedcelllinesforisolationofswineorigininfluenzaavirusesfromdifferentclinicalspecimens AT neilpople comparativesusceptibilityofmadindarbycaninekidneymdckderivedcelllinesforisolationofswineorigininfluenzaavirusesfromdifferentclinicalspecimens AT yohannesberhane comparativesusceptibilityofmadindarbycaninekidneymdckderivedcelllinesforisolationofswineorigininfluenzaavirusesfromdifferentclinicalspecimens |