Validation of ELISAs for the detection of antibodies to Sarcoptes scabiei in pigs.
An Enzyme-linked ImmunoSorbent Assay (ELISA) was developed for the detection of antibodies to Sarcoptes scabiei. This 'Animal Health Service'-ELISA (AHS-ELISA) was compared with a commercial test (Checkit(R) Sarcoptest) using experimental and field sera. The experimental study was a contact infestation experiment. Eighty piglets were randomly divided between the experimental and control group. After introduction of three Sarcoptes scabiei var. suis infested pigs in the experimental group, both groups were monitored by determining scratching indices, taking ear scrapings and blood samples in Weeks 0, 2, 4, 6, 8, 12 and 16. Four pigs in the control group were immunised with either Dermatophagoides pteronyssinus (Dp) antigens (n=2), or Acarus siro (As) antigens (n=2). In the control group all (non-immunised) pigs were negative in all tests. In the experimental group only slightly elevated scratching indices were observed, with a maximum in Week 8. After 2 weeks for the first time an ear scraping was positive (2.5%). In Week 8 the highest number of positive ear scrapings were found (25.0%). Positive results in the Sarcoptest were first obtained in Week 12 (10.5% positive), while eventually 29.0% of the finishing pigs were positive after 16 weeks. The AHS-ELISA first detected a serological response after 6 weeks (5. 0% positives), increasing until after 16 weeks a large proportion (74.2%) of the finishing pigs were seropositive, making the AHS-ELISA the most sensitive test. In the AHS-ELISA one As-immunised pig remained seronegative, but the other hyper-immunised pigs crossreacted. In the Sarcoptest, only Dp-immunised pigs had elevated Optical Densities (OD's) albeit below the cut-off level. Although hyper-immunisation is not a representation of field conditions, it cannot be excluded that the AHS-ELISA is not 100% specific.Field samples were taken from 20 sows in 30 herds, classified as mange-free, suspect, or infested. On a herd level there was high agreement among the ELISAs. Both serological tests were suitable to distinguish mange-free herds from infested herds. In one infested herd the decline of maternal antibody in piglets was studied by sampling 40 piglets from 20 different litters. The lowest average OD using the AHS-ELISA was found at 5 weeks of age, followed by a significant increase at 7 weeks. The average OD with the Sarcoptest was at a minimum level at 3 weeks, but no increase was found later. For screening of herds, interference of maternal antibodies is avoided by sampling at an age of 7 weeks or older.
van der Heijden
H M
HM
R&D, Animal Health Service in the Netherlands, P.O. Box 9, 7400 AA, Deventer, The Netherlands. h.vd.heijden@gdvdieren.nl
Rambags
P G
PG
Elbers
A R
AR
van Maanen
C
C
Hunneman
W A
WA
D000818:Animals; D000906:Antibodies / Q000032:analysis*; D039741:Antigens, Dermatophagoides; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods / Q000662:veterinary*; D006023:Glycoproteins / Q000276:immunology; D012520:Sarcoptes scabiei / Q000276:immunology*; D012532:Scabies / Q000276:immunology / Q000662:veterinary*; D013552:Swine; D013553:Swine Diseases / Q000276:immunology*
D000906:Antibodies; D039741:Antigens, Dermatophagoides; D006023:Glycoproteins