Premium
Testing of the influenza virus purification by CIEF
Author(s) -
Horká Marie,
Kubíček Oldřich,
Kubesoví Anna,
Kubíčková Zuzana,
Rosenbergová Kateřina,
Šlais Karel
Publication year - 2010
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200900310
Subject(s) - embryonated , virus , chromatography , virology , influenza a virus , biology , immunomagnetic separation , chemistry , microbiology and biotechnology
Abstract In virological practice, the pre‐concentration, purification and subsequent determination of the purity and concentration of the viruses from the cultural medium and/or from the real sample are required. The conventional techniques used today are equipment demanding, time‐consuming and laborious. In this study, the CIEF of influenza viruses with UV detection has been developed and subsequently used to test the purification of the virus from the biological samples. The equine and swine influenza viruses present in infected allantoic fluid of specific pathogen free embryonated chicken eggs were precipitated by using PEG 6000 and sodium chloride. The precipitated viruses were centrifuged at 14 000× g , and the impurities of different densities were removed by using the sucrose gradients. The efficiency of the virus purification technique was examined by the CIEF and compared to the results of real‐time PCR. The p I s of both influenza viruses were determined. Simultaneously, the CIEF was found to be a suitable method for the rapid testing of the efficiency of the virus purification.