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J Virol Methods. 2011 Nov 3. [Epub ahead of print]
Standardization of denaturing gradient gel electrophoresis (DGGE) for discrimination of Victoria and Yamagata lineages of influenza B.
Resende PC, Siqueira MM, Born PS, Motta FC.
Source
Instituto Oswaldo Cruz, FIOCRUZ, Laborat?rio de V?rus Respirat?rios e do Sarampo, Pav. H?lio e Peggy Pereira, Av. Brasil 4365, 21040-360 Rio de Janeiro, Brazil.
Abstract
In this study, a PCR-DGGE protocol was standardized in order to distinguish Victoria and Yamagata influenza B lineages directly from clinical samples. After routine multiplex PCR characterization, amplicons of the haemagglutinin gene bearing a 40bp-length GC clamp were generated by nested-PCR and analyzed by electrophoresis in 6% polyacrylamide gel with a 25-45% urea-formamide gradient. The results showed a perfect correlation between DGGE and phylogenetic analyses for all compared samples, besides some distinct profiles in Victoria and Yamagata groups that could be used to infer variability inside these groups. In summary, this DGGE protocol for the haemagglutinin gene is rapid, useful and efficient, being an alternative for discrimination between the influenza B lineages.
Copyright ? 2011. Published by Elsevier B.V.
PMID:
22079619
[PubMed - as supplied by publisher]
Standardization of denaturing gradient gel electrophoresis (DGGE) for discrimination of Victoria and Yamagata lineages of influenza B.
Resende PC, Siqueira MM, Born PS, Motta FC.
Source
Instituto Oswaldo Cruz, FIOCRUZ, Laborat?rio de V?rus Respirat?rios e do Sarampo, Pav. H?lio e Peggy Pereira, Av. Brasil 4365, 21040-360 Rio de Janeiro, Brazil.
Abstract
In this study, a PCR-DGGE protocol was standardized in order to distinguish Victoria and Yamagata influenza B lineages directly from clinical samples. After routine multiplex PCR characterization, amplicons of the haemagglutinin gene bearing a 40bp-length GC clamp were generated by nested-PCR and analyzed by electrophoresis in 6% polyacrylamide gel with a 25-45% urea-formamide gradient. The results showed a perfect correlation between DGGE and phylogenetic analyses for all compared samples, besides some distinct profiles in Victoria and Yamagata groups that could be used to infer variability inside these groups. In summary, this DGGE protocol for the haemagglutinin gene is rapid, useful and efficient, being an alternative for discrimination between the influenza B lineages.
Copyright ? 2011. Published by Elsevier B.V.
PMID:
22079619
[PubMed - as supplied by publisher]
