1.
Open Session of the EuFMD - Cascais –Portugal 26-28 October 2016
TAILED PRIMERS ENHANCE REAL-
TIME RT-PCR DETECTION OF FOOT
AND MOUTH DISEASE VIRUS
F. Vandenbussche, E. Mathijs, D. Lefebvre,
K. De Clercq and S. Van Borm

2.
Use of a portable real-time reverse transcriptase-polymerase chain reaction assay for
rapid detection of foot-and-mouth disease virus.
Callahan JD, Brown F, Osorio FA, Sur JH, Kramer E, Long GW, Lubroth J, Ellis SJ,
Shoulars KS, Gaffney KL, Rock DL, Nelson WM.
J Am Vet Med Assoc. 2002 Jun 1;220(11):1636-42.
3D
5’-UTR
3D and
5’-UTR
Reference methods

3.
But what about specificity?
When your authorities demand you to demonstrate that a sample
does NOT contain FMDV…

4.
But what about specificity?
When your authorities demand you to demonstrate that a sample
does NOT contain FMDV…
• Contamination ?
• Cross-reactivity ?
• PCR artefact ?
• …
• Small amount of FMD
viral RNA ?

5.
But what about specificity?
When your authorities demand you to demonstrate that a sample
does NOT contain FMDV…
• Contamination ?
• Cross-reactivity ?
• PCR artefact ?
• …
• Small amount of FMD
viral RNA ?
To improve the robustness and overall performance of the assays ?!

6.
5’-UTR: dilution series with tailed and non-tailed primers

7.
5’-UTR: effect of tailing of forward and/or reverse primers

8.
3D: 5 / 50 FMDV strains showed Cq value shift (3.43 ± 0.11)

9.
3D: 5 / 50 FMDV strains showed Cq value shift (3.43 ± 0.11)

10.
Thank you for your attention!