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Testing of the influenza virus purification by CIEF
M. Horká, O. Kubíček, A. Kubesová, Z. Kubíčková, K. Rosenbergová, K. Šlais
Language English Country Germany
Document type Comparative Study, Journal Article, Research Support, Non-U.S. Gov't
- MeSH
- Isoelectric Focusing methods MeSH
- Isoelectric Point MeSH
- Hydrogen-Ion Concentration MeSH
- Horses MeSH
- Virus Cultivation methods MeSH
- Orthomyxoviridae chemistry growth & development isolation & purification MeSH
- Reverse Transcriptase Polymerase Chain Reaction methods MeSH
- Swine MeSH
- Sucrose MeSH
- Animals MeSH
- Check Tag
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Comparative Study MeSH
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 x 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 pIs 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.
References provided by Crossref.org
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