Application of a flow type quartz crystal microbalance immunosensor for real time determination of cattle bovine ephemeral fever virus in liquid.

Bovine ephemeral fever (BEF) is a viral disease of cattle. A flow type quartz crystal microbalance (QCM) immunosensor was developed for the real time determination BEF virus (BEFV) that is suitable for clinical point-case diagnosis. Self-assembled monolayer (SAM) of thiols and sulphides by the cystamine-glutaraldehyde method was used for the immobilization of BEFV monoclonal antibody on the gold surface of a quartz crystal microbalance (QCM). A positive correlation was found between the virus concentration and frequency changes (R(2)=0.9962) on this QCM system. The reproducible rates for the 50 and 10mug/mL samples were 4 and 13.9%, respectively. There was no interference from non-specifically adsorbed phage. Using this flow type QCM immunosensor, BEFV could specifically be detected with sensitivity comparable to a conventional enzyme-linked immunosorbent assay (ELISA). The measurement could be obtained directly, within several minutes, rather than hours as required visualizing the results of ELISA. In addition, the observation of reproducible and constant changes after successive additions of BEFV suggests that a QCM immunosensor in a flow cell could be developed for automated or continuous real time operation.