Table 2.
Summary of methodologies used to detect evidence of contact with disease agents in carnivores in the Iberian lynx distribution areas during 2004–2006, showing samples analysed, threshold for serology (if quantitative), sequences used for polymerase chain reaction (PCR), conjugates used for immunofluorescence, and media used for culture
| Agenta | Sampleb | Assayc | Detectsd | Name of commercial kits, positive cut-off (serology), sequences (PCR), conjugates (FA), and media (culture) | Manufacturers (in parenthesis) and/or references |
|---|---|---|---|---|---|
| FCV | Serum | cELISA | c | F1008-AB02 Feline Calicivirus ELISA | Eurovet Veterinaria |
| FHV-1 | Serum | cELISA | c | F107-AB02 Feline Herpes ELISA | Eurovet Veterinaria |
| Oro-pharyngeal swab | FA | ai | Feline serum anti-FHV-1 | Eurovet Veterinaria | |
| 210-41-FHV/FVR | VMRD | ||||
| FCoV | Serum | cELISA | c | Ingezim Corona Felino | Ingenasa |
| Enteric mucosa, mesenteric LN, faeces | FA | ai | Feline serum anti-FCoV | Ingenasa | |
| Anti-felineIgG FITC conjugate | VMRD | ||||
| RIC | ai | FCoVMegacor (FB995) | VMRD | ||
| FeLV | Serum | cELISA | c | Ingezim FeLV | Ingenasa |
| Serum | ELISA-d | ai | Ingezim FeLV-Das | Ingenasa | |
| FIV | Serum | cELISA | c | Ingezim FIV | Ingenasa |
| PV | Serum | cELISA | c | Ingezim CPV | Ingenasa |
| Intestine, intestinal content, faeces | PCR | ai | BAD 1017A | Eurokit S.L. | |
| VPF (5’-GAT GGC ACC TCC GGC AAA GA-3’) | Reed et al. (1988) | ||||
| VPR (5’-TTT CTA GGT GCT AGT TGA G-3’) | |||||
| ELISA-d | ai | Ingezim CPV/FPV-Das | Ingenasa | ||
| IPo | ai | Rabbit IgG anti-feline panleukopenia virus 29989 ImmunoPure biotinylated protein A. 21123 ImmunoPure Avidin, Horseradish peroxidase conjugate | Pierce Chemical Co., Palacín, 1984, Rakich et al., 1986 | ||
| FA | ai | Feline serum anti-FPV | Ingenasa | ||
| Anti-feline IgG FITC conjugate | VMRD | ||||
| CDV | Serum | cELISA | c | Ingezim Moquillo | Ingenasa |
| Tonsils, retro-pharyngeal and/or submandibular LN | FA | ai | Canine serum anti-CDV | Ingenasa | |
| Anti-canine IgG FITC conjugate | VMRD | ||||
| PCR | ai | Ingene CDV | Ingenasa | ||
| CAV-1 | Serum | cELISA | c | Canine hepatitis IgG ELISA | Eurovet Veterinaria |
| Tonsils, retro-pharyngeal and/or submandibular LN | FA | ai | Canine serum anti-CAV-1 | Eurovet Veterinaria | |
| Anti-canine IgG FITC conjugate | VMRD | ||||
| EH | Serum | cELISA | c | Ingezim Ehrlichia | Ingenasa |
| Spleen | FA | ai | Canine serum positive to Ehrlichia canis. | VMRD | |
| Ref-211-P-EC Rabbit anti-dog IgG Fc fragment antibody, FITC conjugated | Rockland Immunochemicals | ||||
| CH | Serum | cELISA | c | Chlamydia antibodies | Eurovet Veterinaria |
| Conjunctival swab | RIC | ai | Clearview Chlamydia MF | Unipath | |
| LI | Serum | MAL | c | 1:100 | Faine (1982) |
| Kidney and/or urine | Staining | ai | Argentic stain | León-Vizcaíno et al. (1987) | |
| IPo + IFA | ai | Ig Rabbit anti Leptospira interrogans serovar Goat Ig anti-rabbit IgG: GAR/Ig/FITC or /IPo | Nordic Immunological Laboratories, León-Vizcaíno et al. (1987) | ||
| Culture | ai | EMJH medium | Difco Laboratories Baranton and Postic (1989) | ||
| BB | Tonsil smears | FA | ai | Serum anti Borrelia burgdorferi | Institut Pasteur Production VMRD, Baranton and Postic (1989) |
| Anti-canine IgG FITC | |||||
| SE | Multiorgan | Culture | ai | Selenite-cystine and Rappaport-Vassiliadis + Salmonella-Shigella agar and brilliant green agar | Millán et al., 2004 |
| MB | Serum | cELISA | c | Protein MBP70; ⩾30% | Martín-Atance et al., 2006 |
| Multiorgan | Culture | ai | Middlebrook’s 7H11 and Lowenstein-Jensen media | Corner and Nicolacopoulos, 1988, Gallager and Horwill, 1977 | |
| PCR | ai | TB1-F (5´-GAA CAA TCC GCA GTT GAC AA-3´) | Böddinghauss et al., 1990, Liébana et al., 1996 | ||
| TB1-R (5´-AGC ACG CTG TCA ATC ATG TA-3´) | |||||
| TG | Serum | MAT | c | 1:25 | Dubey and Desmonts (1987) |
| Faeces | FA | ai | Toxo-Troll ‘F’ 75 411 | BioMérieux Industry | |
| NC | Serum | ELISA + IFA | c | >40% | VMRD |
| CF | Entire blood | PCR | ai | 7549 (5´-GTCAGGATCCTGGGTTGATCCTGCCAG-3´) | Reichard et al., 2005, Millán et al., 2007a |
| 7548 (5´-GACTGAATTCGACTTCTCCTTCCTTTAAG-3´) |
FCV, feline calicivirus; FHV-1, feline herpesvirus-1; FCoV, feline coronavirus; FeLV, feline leukemia virus; FIV, feline immunodeficiency virus; PV, feline parvovirus subgroup virus; CDV, canine distemper virus; CAV-1, canine adenovirus-1; EH, Ehrlichia spp.; CH, Chlamydophila spp.; LI, Leptospira interrogans (serovars Australis, Autumnalis, Ballum, Bataviae, Bratislava, Canicola, Grippotyphosa, Hardjo, Hebdomadis, Icterogaemorrhagiae, Pomona, Saxkoebing, Sejroë, Tarassovi); BB, Borrelia burgdorferi; SE, Salmonella enterica; MB, Mycobacterium bovis; TG, Toxoplasma gondii; NC, Neospora caninum; CF, Cytauxzoon felis.
LN, lymph node.
ELISA, enzyme-linked immunosorbent assay (c, competitive; d, direct); FA, direct immunofluorescence; IFA, indirect immunofluorescence assay; IPo, immunoperoxidase; LA, latex aglutination; MAL, microaglutination-lisis; MAT, modified agglutination test; RIC, rapid immunochromatography.
ai, active infection; c, contact with the agent.