Multiplexed serotype-specific real time PCR assays -- a valuable tool to
support large scale surveillance for bluetongue virus infection.
- Peter Kirkland,
- B. Farrugia,
- M.J. Frost,
- C. Zhang,
- Deborah Finlaison
Deborah Finlaison
Elizabeth Macarthur Agricultural Institute
Author ProfileAbstract
In the last decade, real time PCR has been increasingly adopted for
bluetongue diagnosis with both broadly reactive and serotype-specific
assays widely used. The use of these assays and nucleic acid sequencing
technologies have enhanced bluetongue virus detection, resulting in the
identification of a number of new serotypes. As a result, more than 30
different serotypes are proposed. Rapid identification of the virus
serotype is essential for matching of antigens used in vaccines and to
undertake surveillance and epidemiological studies to assist risk
management. However, it is not uncommon for multiple serotypes to
circulate in a region either concurrently or in successive years. It is
therefore necessary to have a large suite of assays available to ensure
that the full spectrum of viruses is detected. Nevertheless, covering a
large range of virus serotypes is demanding from both a time and
resource perspective. To overcome these challenges, real time PCR assays
were optimised to match local virus strains and then combined in a panel
of quadriplex assays, resulting in 3 assays to detect 12 serotypes
directly from blood samples from cattle and sheep. These multiplex
assays have been used extensively for bluetongue surveillance in both
sentinel animals and opportunistically collected samples. A protocol to
adapt these assays to capture variations in local strains of bluetongue
virus and to expand the panel is described. Collectively these assays
provide powerful tools for surveillance and the rapid identification of
bluetongue virus serotypes directly from animal blood samples.28 Jan 2022Submitted to Transboundary and Emerging Diseases 30 Jan 2022Submission Checks Completed
30 Jan 2022Assigned to Editor
07 Feb 2022Reviewer(s) Assigned
25 Feb 2022Review(s) Completed, Editorial Evaluation Pending
26 Feb 2022Editorial Decision: Revise Minor
21 Apr 20221st Revision Received
21 Apr 2022Submission Checks Completed
21 Apr 2022Assigned to Editor
17 May 2022Reviewer(s) Assigned
23 May 2022Review(s) Completed, Editorial Evaluation Pending
23 May 2022Editorial Decision: Accept