1. 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. J Am Vet Med Assoc. 2002 Jun 1;220(11):1636-42.
Reference methods
5’-UTR 3D
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 Jun 1;220(11):
3D and 5’-UTR

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 ?!

7. 5’-UTR: dilution series with tailed and non-tailed primers

8. 5’-UTR: effect of tailing of forward and/or reverse primers

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

10. 3D: 5 / 50 FMDV strains showed Cq value shift (3.43 ± 0.11)

12. Thank you for your attention!