Table 2.
Clinical signs, specific antibody reactivity, DTH test results against total L. infantum antigen, Ni reagent, and recombinant Leishmania proteins rKMPII, rTRYP, rLACK, and rpapLe22, and qPCR in the blood of 27 dogs in Mallorca, Spain
| ID | Signs | ELISA* | DTH† | qPCR | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| CTLA | KMP11 | TRYP | LACK | papLe22 | LST | Ni | KMPII + Ni | TRYP + Ni | LACK + Ni | papLe + Ni | Blood‡ | ||
| M1 | − | 0.017 | 0.000 | 0.000 | 0.000 | 0.000 | 5.5§ | 0.0 | 0.0 | 5.5 | 0.0 | 0.0 | 131.2 |
| M2 | − | 0.063 | 0.000 | 0.000 | 0.000 | 0.000 | 0.0 | 8.5 | 7.5 | 9.5 | 4.0 | 0.0 | 0.0 |
| M3 | − | 0.035 | 0.000 | 0.000 | 0.000 | 0.000 | 0.0 | 6.5 | 0.0 | 0.0 | 0.0 | 0.0 | 34.7 |
| M4 | − | 0.045 | 0.000 | 0.000 | 0.000 | 0.000 | 13.0§ | 0.0 | 0.0 | 5.5 | 0.0 | 0.0 | 1.0 |
| M5 | − | 0.036 | 0.000 | 0.000 | 0.000 | 0.000 | 8.0 | 8.5 | 14.0 | 7.5 | 6.0 | 6.5 | 0.0 |
| M6 | − | 0.039 | 0.000 | 0.000 | 0.000 | 0.000 | 7.5 | 5.0 | 10.5 | 15.5 | 11.5 | 12.5 | 1.0 |
| M7 | + | 0.260 | 0.000 | 0.000 | 0.000 | 0.000 | 10.5§ | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 1.0 |
| M8 | − | 0.133 | 0.000 | 0.000 | 0.000 | 0.000 | 8.5 | 5.0 | 8.5 | 15.0 | 0.0 | 0.0 | 22.8 |
| M9 | − | 0.08 | 0.000 | 0.000 | 0.000 | 0.000 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 3.5 |
| M10 | − | 0.015 | 0.000 | 0.000 | 0.000 | 0.000 | 0.0 | 0.0 | 0.0 | 5.5 | 0.0 | 0.0 | 0.0 |
| M11 | − | 0.031 | 0.000 | 0.000 | 0.000 | 0.000 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 |
| M12 | + | 0.330 | 0.000 | 0.000 | 0.000 | 0.000 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 |
| M13 | − | 0.037 | 0.000 | 0.000 | 0.000 | 0.000 | 0.0 | 0.0 | 0.0 | 6.0 | 0.0 | 6.0 | 14.0 |
| M14 | − | 0.028 | 0.000 | 0.000 | 0.000 | 0.000 | 6.5§ | 0.0 | 12.0 | 0.0 | 0.0 | 7.0 | 1.9 |
| M15 | − | 0.141 | 0.000 | 0.000 | 0.000 | 0.000 | 14.0§ | 0.0 | 0.0 | 12.5 | 6.0 | 0.0 | 2.2 |
| M16 | + | 1.860 | 0.000 | 0.000 | 0.000 | 0.369 | 6.5§ | 0.0 | 6.0 | 0.0 | 0.0 | 0.0 | 0.0 |
| M17 | − | 0.043 | 0.000 | 0.000 | 0.000 | 0.000 | 8.0 | 6.5 | 8.5 | 17.5 | 12.0 | 6.0 | 7.0 |
| M18 | − | 0.013 | 0.000 | 0.000 | 0.000 | 0.000 | 0.0 | 0.0 | 0.0 | 0.0 | 5.5 | 7.5 | 0.0 |
| M19 | + | 3.000 | 1.737 | 0.000 | 0.000 | 0.255 | 5.5§ | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 45.7 |
| M20 | − | 0.047 | 0.000 | 0.000 | 0.000 | 0.000 | 5.5§ | 0.0 | 8.0 | 0.0 | 0.0 | 0.0 | 0.0 |
| M21 | − | 0.057 | 0.000 | 0.000 | 0.000 | 0.000 | 8.0 | 5.0 | 0.0 | 0.0 | 0.0 | 13.0 | 0.0 |
| M22 | − | 0.079 | 0.000 | 0.000 | 0.000 | 0.000 | 9.0 | 5.0 | 0.0 | 13.0 | 15.0 | 0.0 | 0.0 |
| M23 | − | 0.061 | 0.000 | 0.000 | 0.000 | 0.000 | 16.5 | 5.0 | 0.0 | 0.0 | 6.5 | 8.0 | 0.0 |
| M24 | + | 0.330 | 0.000 | 0.000 | 0.000 | 0.000 | 8.0§ | 0.0 | 0.0 | 8.5 | 0.0 | 0.0 | 0.0 |
| M25 | − | 0.032 | 0.000 | 0.000 | 0.000 | 0.000 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 | 0.0 |
| M26 | − | 0.024 | 0.000 | 0.000 | 0.000 | 0.000 | 5.5§ | 0.0 | 0.0 | 7.0 | 0.0 | 0.0 | 0.0 |
| M27 | − | 0.037 | 0.000 | 0.000 | 0.000 | 0.000 | 10.0§ | 0.0 | 15.0 | 6.0 | 0.0 | 0.0 | 13.7 |
| n | 5 | 5 | 1 | 0 | 0 | 2 | 18 | 4 | 8 | 2 | 3 | 13 | |
CTLA = crude total L. infantum antigen; LST = leishmanin skin test.
ELISA results are expressed as optical densities (OD). OD results above the cut-off values were considered positive (numbers in bold). The cut-off values (mean + 3 SD for 76 dogs from non-endemic area) were established at 0.180 OD for CTLA, 0.100 OD for rKMPII ELISA, 0.082 OD for rTRYP ELISA, 0.060 OD for rpapLe22 ELISA, and 0.005 OD for rLACK ELISA.
Mean millimeter of the largest perpendicular diameter of induration and eritamatous area and its perpendicular one at 48 and 72 hours post-injection. DTH responses > 5 mm were considered positive (numbers in bold).
Results expressed as x-fold more DNA copies than the calibrator sample (dog M7).
Dogs with a positive LST result and no reaction to Ni reagent. Ni reagent is raw non-infected larva extract in pyrogen-free saline solution.