Rapid separation and identification of the subtypes of swine and equine influenza A viruses by electromigration techniques with UV and fluorometric detection

• Published in: Analyst 2011-07, Vol.136 (14), p.3010-3015
• Main Authors: HORKA, Marie, KUBICEK, Oldřich, KUBESOVA, Anna, ROSENBERGOVA, Kateřina, KUBICKOVA, Zuzana, SLAIS, Karel
• Format: Article
• Language: English
• Subjects: Analytical chemistry; Animals; Chemistry; Chromatographic methods and physical methods associated with chromatography; Electrophoresis, Capillary - methods; Ethers - chemistry; Exact sciences and technology; Fluorescent Dyes - chemistry; Fluorometry; Horses; Hydrogen-Ion Concentration; Influenza A virus - classification; Influenza A virus - isolation & purification; Influenza A Virus, H1N1 Subtype - isolation & purification; Influenza A Virus, H1N2 Subtype - isolation & purification; Influenza A Virus, H3N8 Subtype - isolation & purification; Influenza A Virus, H7N7 Subtype - isolation & purification; Isoelectric Focusing - methods; Other chromatographic methods; Polyvinyls - chemistry; Spectrometric and optical methods; Spectrophotometry, Ultraviolet; Swine
• ISSN: 0003-2654; 1364-5528
• DOI: 10.1039/c0an00896f

Influenza A is viral disease, which is a cause of yearly epidemics and, potentially, pandemics. The conventional techniques used today are equipment-demanding, time-consuming and laborious. Recently, we have confirmed that the capillary isoelectric focusing is a suitable fast alternative for the verifying of virus purity. In the wide pH gradient of pH range 2.0-7.5 the isoelectric points for subtypes of equine (H3N8) and swine (H1N2) influenza A viruses were determined approximately as 6.6 and 6.5, respectively. In this contribution we have verified these findings using different isolates of different viral subtypes of swine influenza, H1N1, H1N2, and of equine influenza, H3N8, H7N7, which were separated by capillary zone electrophoresis (CZE) and capillary isoelectric focusing (CIEF) in the narrow pH gradient pH range from 6.0 to 7.0. It was found that the isoelectric points of different isolates and subtypes of equine and swine influenza are almost independent of their origin. The electromigration velocities of subtypes of equine or swine influenza viruses were dependent on the antigenic subtypes of their surface glycoproteins. The detection sensitivity of the influenza viruses labeled by the fluorescent non-ionogenic tenside based on poly(ethylene glycol)pyrenebutanoate for fluorometric detection was increased and down to ten labeled viruses were detected. The isoelectric points of the native and labeled equine and swine influenza A viruses and their subtypes do not differ. According to our experiments these methods appear to be useful for the fast preliminary differentiation of influenza viruses in future.