Table 3.
Clinical samples used for validation of the FMDV RT‐iiPCR assay
| Species | Animal #/Pen # | Virus strain | DPI | Sample type | Extraction methods |
|---|---|---|---|---|---|
| Cattle | 1 | FMDV O/UKG11/2001 | 1,2,3 | Nasal Swabs | P/M |
| Oral swabs | P/M/t | ||||
| 2 | FMDV O/UKG11/2001 | 1,2,3 | Nasal swabs | P/M | |
| Oral swabs | P/M/t | ||||
| Sheep | 535 | FMDV O/UKG11/2001 | 1,2,3 | Nasal and Oral swabs | P/M |
| 566 | FMDV O/UKG11/2001 | 1,2,3 | Nasal Swab | P/M | |
| Oral swabs | P/M/t | ||||
| Pig | 3 | Uninfected | – | Tongue and soft palate epithelium | t |
| 4 | FMDV O/UKG11/2001 | 4 | Vesicular fluid | None | |
| 5 | SVDV PO1/2003 | 17 | Vesicular fluid | None | |
| 14 | FMDV O/UKG11/2001 | 3 | Tongue epithelium | t | |
| 15 | FMDV O/UKG11/2001 | 4 | Vesicular fluid | None | |
| 16 | FMDV O/UKG11/2001 | 3 | Tongue epithelium | t | |
| 24 | FMDV O/UKG11/2001 | 7 | Vesicular fluid | None | |
| 76 | FMDV SAT1/ZAM9/2000 | 1 | Coronary band and interdigital epithelium | M | |
| 77 | FMDV A/IRN1/2005 | 1 | Coronary band and interdigital epithelium | M | |
| 78 | FMDV A/IRN1/2009 | −2, 1, 2, 3, 4 | Nasal and oral swabs | P/M | |
| 79 | FMDV A/IRN1/2009 | −2, 1, 2, 3, 4 | Nasal and oral swabs | P/M | |
| 80 | FMDV A/IRN1/2009 | −2, 1, 2, 3, 4 | Nasal swab | P/M | |
| Oral swab | P/M/t | ||||
| Lesion swab | t | ||||
| 81 | FMDV SAT1/ZAM9/2008 | 6 | Vesicular fluid | None | |
| Pen #1 | FMDV O/UKG11/2001 | 0, 1, 2, 3 | Oral Fluid | M | |
| Pen # 2 | FMDV O/UKG11/2001 | 0, 1, 2, 4 | Oral Fluid | M | |
| Pen # 3 | FMDV O/UKG11/2001 | 0, 1, 2, 5 | Oral Fluid | M | |
| Pen # 4 | FMDV O/UKG11/2001 | 0, 1, 2, 6 | Oral Fluid | M |
A large collection of archived clinical samples (swabs, epithelial tissues and vesicular and oral fluids), collected from cattle, sheep and pigs infected with different FMDV strains, was used for clinical validation of the FMDV RT‐iiPCR assay. Nucleic acid was extracted from the samples using routine laboratory‐based MagMAX™ (M) extraction system, or PetNAD™ (P) column‐based or automated taco™ mini extraction system (t) recommended for the portable POCKIT analyser. Vesicular fluid was diluted in nuclease‐free water and used directly in the FMDV RT‐iiPCR assay. In addition to the samples listed, 10 additional nasal and oral samples from heathy cattle and sheep were used as negative controls for PetNAD™ and MagMAX™ extraction followed by testing in FMDV RT‐iiPCR and FMDV RRT‐PCR assays.
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