RESUMO
An antigen detection kit (Trichin-L), based on latex agglutination and developed by the Bio-Rad company was validated at five European laboratories. The validation parameters included specificity, sensitivity, robustness and reproducibility. Specificity was evaluated by testing parasite antigens from five non-Trichinella parasites in addition to the Trichinella genus. To evaluate sensitivity, 10 pork samples spiked with 1, 3, 6 or 15 Trichinella larvae were tested in each laboratory. To evaluate the robustness of the test, the solubilized antigens were maintained at room temperature and tested at different times. Reproducibility was assessed in each laboratory using 40, 100g minced pork samples, each spiked with Trichinella spiralis. The use of larval homogenates obtained from the Trichin-L kit as a template for parasite identification at the species level by a multiplex PCR, was also evaluated. The results showed a high specificity and sensitivity where solubilized antigens maintained their stability and reactivity for up to three days. Reproducibility was high, as similar results were obtained in the five laboratories. The larval homogenates obtained using the Trichin-L kit were successfully used in multiplex PCRs to identify Trichinella species.
Assuntos
Antígenos de Helmintos/imunologia , Testes de Fixação do Látex/veterinária , Carne/parasitologia , Doenças dos Suínos/diagnóstico , Trichinella spiralis/isolamento & purificação , Triquinelose/veterinária , Animais , Europa (Continente) , Larva , Reação em Cadeia da Polimerase Multiplex/veterinária , Kit de Reagentes para Diagnóstico , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Especificidade da Espécie , Suínos , Doenças dos Suínos/parasitologia , Trichinella spiralis/imunologia , Triquinelose/diagnóstico , Triquinelose/parasitologiaRESUMO
Sarcoptic mange occurs in many parts of the world and is common in populations of domestic and wild canids, including red foxes (Vulpes vulpes). In recent years, an indirect antibody enzyme-linked immunosorbent assay (ELISA), with higher sensitivity and specificity than traditional diagnostic methods, has been successfully applied in the diagnosis of sarcoptic mange in dogs. The same ELISA has also demonstrated specific antibodies to Sarcoptes scabiei in experimentally infected red foxes. The aim of this study was to evaluate the indirect ELISA when used to detect antibodies to S. scabiei in field sera from Swedish red foxes. One cohort of both infected and non-infected red foxes (cohort 1; n = 88), and one cohort of apparently non-infected foxes (cohort 2; n = 67) were examined for skin lesions and presence of S. scabiei by thorough visual examination at autopsy and skin scrapings. Samples of blood-tinted body liquid from the abdomen or thorax cavity were collected and analysed by the indirect ELISA. The relative sensitivity and specificity of the ELISA at different cut-offs (OD values) were estimated by comparing the test results to the infection status as determined by examination and skin scrapings. The highest combination of relative sensitivity and specificity, calculated based on cohort 1, was 95.4 and 100.0%, respectively. These estimates were constant for cut-offs 0.150-0.225, which included the cut-off based on the mean plus three standard deviations of test results from cohort 2 (0.165). It is concluded that this test can be useful in diagnosis and epidemiological studies of S. scabiei infection in red foxes.
Assuntos
Ensaio de Imunoadsorção Enzimática/veterinária , Raposas , Sarcoptes scabiei/imunologia , Escabiose/veterinária , Animais , Anticorpos/sangue , Estudos de Coortes , Ensaio de Imunoadsorção Enzimática/métodos , Feminino , Masculino , Valor Preditivo dos Testes , Escabiose/sangue , Escabiose/diagnóstico , SuéciaRESUMO
In a laboratory study to evaluate the efficacy of selamectin for treatment of canine nasal mite infection, 12 purpose-bred beagles were experimentally infected with Pneumonyssoides caninum (P. caninum). Six of the dogs were treated with selamectin applied to the skin of the back at dosages of 6 to 24 mg/kg for three times at 2-week intervals. The remaining six dogs were an untreated control group. At necropsy 39 to 46 days after inoculation, no P. caninum mites were found in any of the treated dogs. In contrast, nasal mites were found in five of the untreated dogs. This difference was statistically significant at P=0.015.
Assuntos
Antiparasitários/uso terapêutico , Doenças do Cão/tratamento farmacológico , Ivermectina/análogos & derivados , Infestações por Ácaros/veterinária , Cavidade Nasal/parasitologia , Animais , Cães , Relação Dose-Resposta a Droga , Feminino , Ivermectina/uso terapêutico , Masculino , Infestações por Ácaros/tratamento farmacológico , Ácaros/efeitos dos fármacos , Ácaros/crescimento & desenvolvimento , Distribuição Aleatória , Resultado do TratamentoRESUMO
Abstract The purpose of this study was to evaluate a serodiagnostic test (enzyme-linked immunosorbent assay; ELISA) for sarcoptic mange in dogs and to characterize the assay antigen, based on the mite Sarcoptes scabiei var. vulpes. The ELISA, applied to sera from 359 dogs suspected of having sarcoptic mange, showed a sensitivity and specificity of 92 and 96%, respectively. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) of the antigen employed in the ELISA revealed polypeptide bands with molecular weights ranging between 14 and 164 kDa. In Western blot analyses antigens of molecular weights between 62 and 64 kDa dominated. Particularly dominant were antigens of 164 and 147 kDa. These were found to have isoelectrical points in the range of 5.7-6.9. Sera from dogs infected with Cheyletiella sp., Demodex canis, Linognathus setosus and Otodectes cynotis, as well as from dogs allergic to fleas, were negative in the ELISA. Résumé- Le but de cette étude est d'évaluer un test sérologique ELISA pour le diagnostic de la gale sarcoptique chez le chien et de caractériser l'antigène révélateur, extrait de l'acarien Sarcoptes scabiei var. vulpes. Le test ELISA, lors d'une étude conduite avec les sérums de 359 chiens suspects de gale sarcoptique a démontré une sensibilité et une spécificité de 92 et 96%, respectivement. L'électrophorèse en gel polyacrilamide dodécyl sulfate de sodium de l'antigène utilisé dans l'ELISA a révélé des bandes polypeptidiques de poids moléculaire compris entre 14 et 164 kDa. Dans l'analyse en Western blot, les antigènes de poids moléculaire compris entre 62 et 164 kDa étaient les plus abondants, notamment ceux de 164 et 147 kDa. Ces derniers ont des points isoélectriques compris entre 5.7 et 6.9. Les sérums de chiens infectés par des Cheyletiella sp. Demodex canis, Linognathus setosus et Otodectes cynotis, ou par des chiens allergiques aux puces, se sont révélés négatifs en ELISA. [Bornstein, S., Thebo, P., Zakrisson, G. Evaluation of an enzyme-linked immunosorbent assay (ELISA) for the serological diagnosis of canine sarcoptic mange (Evaluation d'un test ELISA pour le diagnostic sérologique de la gale sarcoptique canine). Veterinary Dermatology 1996; 7: 21-28.] Resumen El objetivo de este estudio fue el de evaluar una pruba serodiagnóstica (prueba de inmunoadsorción ligada a enzima; ELISA) para la sarna sarcóptica en el perro y caracterizar el antigeno prueba, basado en el ácaro Sarcoptes scabei, var. vulpes. El ELISA, aplicado a sueros de 359 perros sospechosos de padecer sarna sarcóptica, mostró una sensibilidad y especificidad del 92 y 96%, respectivamente. La electroforesis en gel de poliacrilamida dodecil sulfato sódico (SDS-PAGE) del antigeno usado en el ELISA reveló bandas de polipétidos con peso molecular entre 14 y 164 kDa. En el análisis Western blot, predominaron los antigenos de pesos moleculares entre 62 y 164 kDa. Los antigenos entre 164 y 147 kDa fueron especialmente predominantes. Estos tuvieron puntos isoeléctricos entre 5.7 y 6.9. Los sueros de perros infectados por Cheyletiella sp., Demodex canis, Linognathus setosus y Otodectes cynotis, asi como el de perros alérgicos a las pulgas fueron negativos en el ELISA. [Bornstein, S., Thebo, P., Zakrisson, G. Evaluation of an enzyme-linked immunosorbent assay (ELISA) for the serological diagnosis of canine sarcoptic mange (Evaluation de una prueba de immunoadsorcion ligada a enzima (ELISA) para el diagnostico serologico de la sarna sarcóptica canina). Veterinary Dermatology 1996; 7: 21-28.] Zusammenfassung- Ziel dieser Studie war, einen Serodiagnostiktest (Enzyme-Linked-Immunosorbent-Assay, ELISA) für Sarkoptesräude des Hundes zu überprüfen und das Testantigen zu charakterisieren, das auf der Milbe Sarcoptes scabiei var. vulpes basiert. Der ELISA-Test, der bei den Sera von 359 Hunden mit Sarkoptesverdacht angewendet wurde, zeigte eine Sensitivität von 92% bzw. 96%. Die Natriumdodecylsul-fatpolyacrylamid-Gelelektrophorese (SDS-PAGE) des Antigen, das im ELISA verwendet wurde, zeigte Polypeptid-Banden mit Molekulargewichten zwischen 14 und 164 kDa. In der Wester-blot-Analyse dominierten Antigene mit einem Molekulargewicht zwischen 62 und 164 kDa. Besonders dominierend waren Antigene von 164 und 147 kDa. Bei diesen stellte man isoelektrische Punkte im Bereich von 5,7 bis 6,9 fest. Die Sera von Hunden, die mit Cheyletiella sp., Demodex canis, Linognathus setosus und Otodectes cynotis infiziert waren, fielen ebenso wie die Hunde mit Allergie auf Flöhe im ELISA negativ aus. [Bornstein, S., Thebo, P., Zakrisson, G. Evaluation of an enzyme-linked immunosorbent assay (ELISA) for the serological diagnosis of canine sarcoptic mange (Die Auswertung eines Enzym-Linked-Immunosorbent-Assay (ELISA) für die serologische Diagnose der kaninen Sarkoptesräude). Veterinary Dermatology 1996; 7: 21-28.].