Application of real time RT-PCR for the genetic homogeneity and stability tests of the seed candidates for live attenuated influenza vaccine production.
Identifieur interne : 000747 ( Main/Exploration ); précédent : 000746; suivant : 000748Application of real time RT-PCR for the genetic homogeneity and stability tests of the seed candidates for live attenuated influenza vaccine production.
Auteurs : Svetlana Shcherbik [États-Unis] ; Sheila B. Sergent ; William G. Davis ; Bo Shu ; John Barnes ; Irina Kiseleva ; Natalie Larionova ; Alexander Klimov ; Tatiana BousseSource :
- Journal of virological methods [ 1879-0984 ] ; 2014.
Descripteurs français
- KwdFr :
- Animaux, Contrôle de qualité, Embryon de poulet, Humains, Instabilité du génome, Orthomyxoviridae (génétique), Réaction de polymérisation en chaine en temps réel (), Technologie pharmaceutique (), Vaccins antigrippaux (génétique), Vaccins antigrippaux (normes), Vaccins atténués (génétique), Vaccins atténués (normes), Variation génétique, Virologie ().
- MESH :
- génétique : Orthomyxoviridae, Vaccins antigrippaux, Vaccins atténués.
- normes : Vaccins antigrippaux, Vaccins atténués.
- Animaux, Contrôle de qualité, Embryon de poulet, Humains, Instabilité du génome, Réaction de polymérisation en chaine en temps réel, Technologie pharmaceutique, Variation génétique, Virologie.
English descriptors
- KwdEn :
- Animals, Chick Embryo, Genetic Variation, Genomic Instability, Humans, Influenza Vaccines (genetics), Influenza Vaccines (standards), Orthomyxoviridae (genetics), Quality Control, Real-Time Polymerase Chain Reaction (methods), Technology, Pharmaceutical (methods), Vaccines, Attenuated (genetics), Vaccines, Attenuated (standards), Virology (methods).
- MESH :
- chemical , genetics : Influenza Vaccines, Vaccines, Attenuated.
- chemical , standards : Influenza Vaccines, Vaccines, Attenuated.
- genetics : Orthomyxoviridae.
- methods : Real-Time Polymerase Chain Reaction, Technology, Pharmaceutical, Virology.
- Animals, Chick Embryo, Genetic Variation, Genomic Instability, Humans, Quality Control.
Abstract
Development and improvement of quality control tests for live attenuated vaccines are a high priority because of safety concerns. Live attenuated influenza vaccine (LAIV) viruses are 6:2 reassortants containing the hemagglutinin (HA) and neuraminidase (NA) gene segments from circulating influenza viruses to induce protective immune responses, and the six internal gene segments from a cold-adapted Master Donor Virus (MDV). LAIV candidate viruses for the 2012-2013 seasons, A/Victoria/361/2011-CDC-LV1 (LV1) and B/Texas/06/2011-CDC-LV2B (LV2B), were created by classical reassortment of A/Victoria/361/2011 and MDV-A A/Leningrad/134/17/57 (H2N2) or B/Texas/06/2011 and MDV-B B/USSR/60/69. In an attempt to provide better identity and stability testing for quality control of LV1 and LV2B, sensitive real-time RT-PCR assays (rRT-PCR) were developed to detect the presence of undesired gene segments (HA and NA from MDV and the six internal genes from the seasonal influenza viruses). The sensitivity of rRT-PCR assays designed for each gene segment ranged from 0.08 to 0.8EID50 (50% of Egg Infectious Dose) per reaction for the detection of undesired genes in LV1 and from 0.1 to 1EID50 per reaction for the detection of undesired genes in LV2B. No undesired genes were detected either before or after five passages of LV1 or LV2B in eggs. The complete genome sequencing of LV1 and LV2B confirmed the results of rRT-PCR, demonstrating the utility of the new rRT-PCR assays to provide the evidence for the homogeneity of the prepared vaccine candidate.
DOI: 10.1016/j.jviromet.2013.09.003
PubMed: 24056261
Affiliations:
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Le document en format XML
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Live attenuated influenza vaccine (LAIV) viruses are 6:2 reassortants containing the hemagglutinin (HA) and neuraminidase (NA) gene segments from circulating influenza viruses to induce protective immune responses, and the six internal gene segments from a cold-adapted Master Donor Virus (MDV). LAIV candidate viruses for the 2012-2013 seasons, A/Victoria/361/2011-CDC-LV1 (LV1) and B/Texas/06/2011-CDC-LV2B (LV2B), were created by classical reassortment of A/Victoria/361/2011 and MDV-A A/Leningrad/134/17/57 (H2N2) or B/Texas/06/2011 and MDV-B B/USSR/60/69. In an attempt to provide better identity and stability testing for quality control of LV1 and LV2B, sensitive real-time RT-PCR assays (rRT-PCR) were developed to detect the presence of undesired gene segments (HA and NA from MDV and the six internal genes from the seasonal influenza viruses). The sensitivity of rRT-PCR assays designed for each gene segment ranged from 0.08 to 0.8EID50 (50% of Egg Infectious Dose) per reaction for the detection of undesired genes in LV1 and from 0.1 to 1EID50 per reaction for the detection of undesired genes in LV2B. No undesired genes were detected either before or after five passages of LV1 or LV2B in eggs. The complete genome sequencing of LV1 and LV2B confirmed the results of rRT-PCR, demonstrating the utility of the new rRT-PCR assays to provide the evidence for the homogeneity of the prepared vaccine candidate. </div></front></TEI><affiliations><list><country><li>États-Unis</li></country></list><tree><noCountry><name sortKey="Barnes, John" sort="Barnes, John" uniqKey="Barnes J" first="John" last="Barnes">John Barnes</name><name sortKey="Bousse, Tatiana" sort="Bousse, Tatiana" uniqKey="Bousse T" first="Tatiana" last="Bousse">Tatiana Bousse</name><name sortKey="Davis, William G" sort="Davis, William G" uniqKey="Davis W" first="William G" last="Davis">William G. Davis</name><name sortKey="Kiseleva, Irina" sort="Kiseleva, Irina" uniqKey="Kiseleva I" first="Irina" last="Kiseleva">Irina Kiseleva</name><name sortKey="Klimov, Alexander" sort="Klimov, Alexander" uniqKey="Klimov A" first="Alexander" last="Klimov">Alexander Klimov</name><name sortKey="Larionova, Natalie" sort="Larionova, Natalie" uniqKey="Larionova N" first="Natalie" last="Larionova">Natalie Larionova</name><name sortKey="Sergent, Sheila B" sort="Sergent, Sheila B" uniqKey="Sergent S" first="Sheila B" last="Sergent">Sheila B. Sergent</name><name sortKey="Shu, Bo" sort="Shu, Bo" uniqKey="Shu B" first="Bo" last="Shu">Bo Shu</name></noCountry><country name="États-Unis"><noRegion><name sortKey="Shcherbik, Svetlana" sort="Shcherbik, Svetlana" uniqKey="Shcherbik S" first="Svetlana" last="Shcherbik">Svetlana Shcherbik</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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