Isolation of influenza viruses in MDCK 33016PF cells and clearance of contaminating respiratory viruses.
Identifieur interne : 000252 ( Main/Curation ); précédent : 000251; suivant : 000253Isolation of influenza viruses in MDCK 33016PF cells and clearance of contaminating respiratory viruses.
Auteurs : Bernhard Roth [Allemagne] ; Hannah Mohr ; Martin Enders ; Wolfgang Garten ; Jens-Peter GregersenSource :
- Vaccine [ 1873-2518 ] ; 2012.
Descripteurs français
- KwdFr :
- Allemagne, Animaux, Chiens, Co-infection (virologie), Culture virale (), Humains, Infections de l'appareil respiratoire (diagnostic), Infections de l'appareil respiratoire (virologie), Lignée cellulaire, Maladies virales (diagnostic), Maladies virales (virologie), Réaction de polymérisation en chaine multiplex, Virologie (), Virus (croissance et développement), Virus (génétique), Virus (isolement et purification).
- MESH :
- croissance et développement : Virus.
- diagnostic : Infections de l'appareil respiratoire, Maladies virales.
- génétique : Virus.
- isolement et purification : Virus.
- virologie : Co-infection, Infections de l'appareil respiratoire, Maladies virales.
- Allemagne, Animaux, Chiens, Culture virale, Humains, Lignée cellulaire, Réaction de polymérisation en chaine multiplex, Virologie.
- Wicri :
- geographic : Allemagne.
English descriptors
- KwdEn :
- Animals, Cell Line, Coinfection (virology), Dogs, Germany, Humans, Multiplex Polymerase Chain Reaction, Respiratory Tract Infections (diagnosis), Respiratory Tract Infections (virology), Virology (methods), Virus Cultivation (methods), Virus Diseases (diagnosis), Virus Diseases (virology), Viruses (genetics), Viruses (growth & development), Viruses (isolation & purification).
- MESH :
- geographic : Germany.
- diagnosis : Respiratory Tract Infections, Virus Diseases.
- genetics : Viruses.
- growth & development : Viruses.
- isolation & purification : Viruses.
- methods : Virology, Virus Cultivation.
- virology : Coinfection, Respiratory Tract Infections, Virus Diseases.
- Animals, Cell Line, Dogs, Humans, Multiplex Polymerase Chain Reaction.
Abstract
This paper summarizes results obtained by multiplex PCR screening of human clinical samples for respiratory viruses and corresponding data obtained after passaging of virus-positive samples in MDCK 33016PF cells. Using the ResPlexII v2.0 (Qiagen) multiplex PCR, 393 positive results were obtained in 468 clinical samples collected during an influenza season in Germany. The overall distribution of positive results was influenza A 42.0%, influenza B 38.7%, adenovirus 1.5%, bocavirus 0.5%, coronavirus 3.3%, enterovirus 5.6%, metapneumovirus 1.0%, parainfluenza virus 0.8%, rhinovirus 4.1%, and respiratory syncytial virus (RSV) 2.5%. Double infections of influenza virus together with another virus were found for adenovirus B and E, bocavirus, coronavirus, enterovirus and for rhinovirus. These other viruses were rapidly lost upon passages in MDCK 33016PF cells and under conditions as applied to influenza virus passaging. Clinical samples, in which no influenza virus but other viruses were found, were also subject to passages in MDCK 33016PF cells. Using lower inoculum dilutions than those normally applied for preparations containing influenza virus (total dilution of the original sample of ∼10(4)), the positive results for the different viruses turned negative already after 2 or 3 passages in MDCK 33016PF cells. These results demonstrate that, under practical conditions as applied to grow influenza viruses, contaminating viruses can be effectively removed by passages in MDCK cells. In combination with their superior isolation efficiency, MDCK cells appear highly suitable to be used as an alternative to embryonated eggs to isolate and propagate influenza vaccine candidate viruses.
DOI: 10.1016/j.vaccine.2011.11.063
PubMed: 22119922
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pubmed:22119922Le document en format XML
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<front><div type="abstract" xml:lang="en">This paper summarizes results obtained by multiplex PCR screening of human clinical samples for respiratory viruses and corresponding data obtained after passaging of virus-positive samples in MDCK 33016PF cells. Using the ResPlexII v2.0 (Qiagen) multiplex PCR, 393 positive results were obtained in 468 clinical samples collected during an influenza season in Germany. The overall distribution of positive results was influenza A 42.0%, influenza B 38.7%, adenovirus 1.5%, bocavirus 0.5%, coronavirus 3.3%, enterovirus 5.6%, metapneumovirus 1.0%, parainfluenza virus 0.8%, rhinovirus 4.1%, and respiratory syncytial virus (RSV) 2.5%. Double infections of influenza virus together with another virus were found for adenovirus B and E, bocavirus, coronavirus, enterovirus and for rhinovirus. These other viruses were rapidly lost upon passages in MDCK 33016PF cells and under conditions as applied to influenza virus passaging. Clinical samples, in which no influenza virus but other viruses were found, were also subject to passages in MDCK 33016PF cells. Using lower inoculum dilutions than those normally applied for preparations containing influenza virus (total dilution of the original sample of ∼10(4)), the positive results for the different viruses turned negative already after 2 or 3 passages in MDCK 33016PF cells. These results demonstrate that, under practical conditions as applied to grow influenza viruses, contaminating viruses can be effectively removed by passages in MDCK cells. In combination with their superior isolation efficiency, MDCK cells appear highly suitable to be used as an alternative to embryonated eggs to isolate and propagate influenza vaccine candidate viruses.</div>
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<Abstract><AbstractText>This paper summarizes results obtained by multiplex PCR screening of human clinical samples for respiratory viruses and corresponding data obtained after passaging of virus-positive samples in MDCK 33016PF cells. Using the ResPlexII v2.0 (Qiagen) multiplex PCR, 393 positive results were obtained in 468 clinical samples collected during an influenza season in Germany. The overall distribution of positive results was influenza A 42.0%, influenza B 38.7%, adenovirus 1.5%, bocavirus 0.5%, coronavirus 3.3%, enterovirus 5.6%, metapneumovirus 1.0%, parainfluenza virus 0.8%, rhinovirus 4.1%, and respiratory syncytial virus (RSV) 2.5%. Double infections of influenza virus together with another virus were found for adenovirus B and E, bocavirus, coronavirus, enterovirus and for rhinovirus. These other viruses were rapidly lost upon passages in MDCK 33016PF cells and under conditions as applied to influenza virus passaging. Clinical samples, in which no influenza virus but other viruses were found, were also subject to passages in MDCK 33016PF cells. Using lower inoculum dilutions than those normally applied for preparations containing influenza virus (total dilution of the original sample of ∼10(4)), the positive results for the different viruses turned negative already after 2 or 3 passages in MDCK 33016PF cells. These results demonstrate that, under practical conditions as applied to grow influenza viruses, contaminating viruses can be effectively removed by passages in MDCK cells. In combination with their superior isolation efficiency, MDCK cells appear highly suitable to be used as an alternative to embryonated eggs to isolate and propagate influenza vaccine candidate viruses.</AbstractText>
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