Standardization of denaturing gradient gel electrophoresis (DGGE) for discrimination of Victoria and Yamagata lineages of influenza B

P C Resende; M M Siqueira; P S Born; F C Motta

doi:10.1016/j.jviromet.2011.10.018

Standardization of denaturing gradient gel electrophoresis (DGGE) for discrimination of Victoria and Yamagata lineages of influenza B

J Virol Methods. 2012 Jan;179(1):212-6. doi: 10.1016/j.jviromet.2011.10.018. Epub 2011 Nov 3.

Authors

P C Resende¹, M M Siqueira, P S Born, F C Motta

Affiliation

¹ Instituto Oswaldo Cruz, FIOCRUZ, Laboratório de Vírus Respiratórios e do Sarampo, Pav. Hélio e Peggy Pereira, Av. Brasil 4365, 21040-360 Rio de Janeiro, Brazil.

PMID: 22079619
DOI: 10.1016/j.jviromet.2011.10.018

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.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Denaturing Gradient Gel Electrophoresis / standards*
Hemagglutinin Glycoproteins, Influenza Virus / genetics
Humans
Influenza B virus / classification*
Influenza B virus / genetics*
Influenza B virus / isolation & purification
Influenza, Human / virology*
Polymerase Chain Reaction / standards*
Virology / methods*

Substances

Hemagglutinin Glycoproteins, Influenza Virus