Veterinary virologists share avian flu data.
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Biomedical subjects
Dennis Senne
Publications and source records attributed to Dennis Senne.
Development of an internal positive control for rapid diagnosis of avian influenza virus infections by real-time reverse transcription-PCR with lyophilized reagents.
We developed an internal positive control (IPC) RNA to help ensure the accuracy of the detection of avian influenza virus (AIV) RNA by reverse transcription (RT)-PCR and real-time RT-PCR (RRT-PCR). The IPC was designed to have the same binding sites for the forward and reverse primers of the AIV matrix gene as the target amplicon, but it had a unique internal sequence used for the probe site. The amplification of the viral RNA and the IPC by RRT-PCR were monitored with two different fluorescent probes in a multiplex format, one specific for the AIV matrix gene and the other for the IPC. The RRT-PCR test was further simplified with the use of lyophilized bead reagents for the detection of AIV RNA. The RRT-PCR with the bead reagents was more sensitive than the conventional wet reagents for the detection of AIV RNA. The IPC-based RRT-PCR detected inhibitors in blood, kidney, lungs, spleen, intestine, and cloacal swabs, but not allantoic fluid, serum, or tracheal swabs The accuracy of RRT-PCR test results with the lyophilized beads was tested on cloacal and tracheal swabs from experimental birds inoculated with AIV and compared with virus isolation (VI) on embryonating chicken eggs. There was 97 to 100% agreement of the RRT-PCR test results with VI for tracheal swabs and 81% agreement with VI for cloacal swabs, indicating a high level of accuracy of the RRT-PCR assay. The same IPC in the form of armored RNA was also used to monitor the extraction of viral RNA and subsequent detection by RRT-PCR.
Animals↗
Viral and bacterial serology of free-ranging Pacific walrus.
Serum or heparinized plasma samples were obtained between 1994 and 1996 from 20 male and 20 female adult free-ranging Pacific walrus (Odobenus rosmarus divergens) from St. Lawrence Island and Round Island, Alaska. Samples were screened for antibodies to some potentially pathogenic bacteria and viruses. No sample had detectable antibody to Brucella spp. Three of 40 (8%) had low antibody titers to Leptospira interrogans serovars. Phocine distemper virus antibodies were not detected. Serologic responses to one or more caliciviruses (San Miguel sea lion virus 12 or vesicular exanthema of swine serotypes E54, F55, G55, 1934B) were detected in 18% (seven of 40) walrus. Antibodies to one or more subtypes of influenza A virus (H10, N2, N3, N5, N6, N7) were detected in 21% (eight of 38). Periodic screening of free-ranging populations for exposure to infectious diseases has become an important component of bio-monitoring programs to facilitate understanding and detecting trends in marine mammal populations.
Alaska↗