Background
Methods
Alignment and phylogenetic reconstruction
Primer, probe design and assay conditions
Analytical sensitivity and specificity of G9R-NA OPXV rt-PCR assay
Old World viruses | Sample ID | Accession No. | Ortholog | Viral Prep. | 2 ng* |
---|---|---|---|---|---|
Camelpox
| CMLV-V78 I | Not available | Not available | Pure | ND |
Cowpox
| CPXV-Sweden | Not available | Not available | Pure | ND |
Ectromelia
| ECTV-MOS | NC_004105 | 73 | Pure | ND |
Monkeypox
| MPXV-USA | DQ011157 | 84 | Pure | ND |
Orf
| ORFV-NZ2 | DQ184476 | 44 | Pure | ND |
Squirrel Fibroma
| SQFV-Kilham | Not available | 59 | Crude | ND |
Tanapox
| TAV-Kenya | NC_009888 | 63 | Pure | ND |
Taterapox
| TATV-DAH68 | NC_008291 | 90 | Pure | ND |
Vaccinia
| VACV-COP | M35027 | 108 | Pure | ND |
Variola
| VARV-BSH | L22579 | 76 | Pure | ND |
North American viruses
| |||||
Raccoonpox
| RCNV-1964 | FJ807746-54 | 70 | Pure | 17 |
Skunkpox
| SKPV-1978 | FJ807755-63 | 86 | Pure | 17 |
Volepox
| VPXV-1985 | FJ807737-45 | 86 | Pure | 16 |
Animal specimen collection and preparation
Specimen | Day 6 | Day 7 | Day 8 | Total No. of specimens | ||||
---|---|---|---|---|---|---|---|---|
(Inf) | (un) | (inf) | (un) | (inf) | (un) | (inf) | (un) | |
Blood | 4 | 1 | 3 | 1 | 2 | 1 | 9 | 3 |
Oral swab | 6 | 1 | 5 | 1 | 7 | 1 | 18 | 3 |
Ocular swab | 6 | 1 | 8 | 1 | 2 | 1 | 13 | 3 |
Anal swab | 5~ | 1 | 3~ | 1 | 2 | 1 | 13 | 3 |
Spleen | 6 | 1 | 5 | 1 | 2 | 1 | 13 | 3 |
Gonad | 6 | 1 | 5 | 1 | 2 | 1 | 13 | 3 |
Kidney | 6 | 1 | 5 | 1 | 2 | 1 | 13 | 3 |
Lymph node | 6 | 1 | 5 | 1 | 2 | 1 | 13 | 3 |
Supradrenal | 6 | 1 | 4 | 1 | 2 | 1 | 12 | 3 |
Brain | 6 | 1 | 5 | 1 | 2 | 1 | 14 | 3 |
Liver | 6 | 1 | 5 | 1 | 2 | 1 | 14 | 3 |
Lung | 6 | 1 | 5 | 1 | 2 | 1 | 14 | 3 |
Skin lesion | 5 | 1 | 2 | 1 | 10 | 1 | 14 | 3 |
Tognue lesion | 5 | 1 | 2 | 1 | NA | 1 | 7 | 3 |
Intestine lesion | NA | 1 | 2 | 1 | NA | 1 | 2 | 3 |
Bladder | NA | NA | 2 | NA | 1 | NA | 3 | NA |
Pancreas | NA | NA | 1 | NA | 1 | NA | 2 | NA |
Number of positive/tested | 79/80 | 0/15 | 64/66 | 0/15 | 41/41 | 0/15 | 184/187 | 0/45 |
Detection of viable OPXV in animal samples through tissue cell culture techniques
Results
Analytical specificity of G9R-NA OPXV rt-PCR assay
Analytical sensitivity of G9R-NA OPXV rt-PCR assay
Virus | DNA (fg) | 200 | 20 | 10 | 5 | 2.5 | 1.25 | 0.625 |
---|---|---|---|---|---|---|---|---|
Raccoonpox
| Pure | 33 | 36.5 | 37.9 | 38.9 (20) | 39.4 (5) | NT | NT |
Skunkpox
| Pure | 28.9 | 32.7 | 34 | 35 | 36 | 37.8 (18) | 39.2 (11) |
Volepox
| Pure | 30.4 | 33.9 | 35.2 | 36.1 | 37.4 | 38.8 (23) | 39.4 (19) |