Bluetongue virus in wild deer, Belgium, 2005-2008.

To investigate bluetongue virus serotype 8 infection in Belgium, we conducted a virologic and serologic survey on 2,416 free-ranging cervids during 2005-2008. Infection emerged in 2006 and spread over the study area in red deer, but not in roe deer.

Simultaneous detection of bluetongue virus RNA, internal control GAPDH mRNA, and external control synthetic RNA by multiplex real-time PCR.

Bluetongue is an insect-borne disease of domestic and wild ruminants that requires strict monitoring by sensitive, reproducible and robust methods. Real-time reverse transcription polymerase chain reaction (RT-qPCR) analysis has become the method of choice for routine viral diagnosis. As false-negative test results can have serious implications; an internal/external control system should be incorporated in each analysis to detect RT-qPCR failure due to poor sample quality, improper nucleic acid extraction and/or PCR inhibition.

A proposed validation method for automated nucleic acid extraction and RT-qPCR analysis: an example using Bluetongue virus.

This study proposes a validation strategy for an automated extraction procedure, followed by RT-qPCR analysis. To avoid false-negative results, a triplex RT-qPCR was used which detects the target viral RNA, an internal and an external control. The methods to determine the validation parameters such as linearity, efficiency, analytical sensitivity, analytical specificity and intra- and interrun variability are described in detail.

Safety concerns for the potential use of cyanovirin-N as a microbicidal anti-HIV agent.

Based on its antiviral activity profile, cyanovirin-N (CV-N) holds great potential for anti-HIV microbicidal application. However, limited data are available on the possible side-effects of this lectin. A detailed investigation was carried out to obtain better insights in the cytotoxic, inflammatory and (anti)-proliferative properties of CV-N in comparison with several other plant-derived lectins.