RESUMO
Anaplasma marginale infection in cattle (n = 216) in the states of Uttar Pradesh and Uttarakhand, North India was screened by microscopy and nested-polymerase chain reaction (PCR). Two recombinant proteins viz. major surface protein (MSP) 5 and MSP2 of A. marginale were expressed in Escherichia coli and their potential in the detection of antibodies to Anaplasma species in the cattle was evaluated by immunoglobulin G-enzyme linked immunosorbent assay (IgG-ELISA). The MSP5 IgG ELISA results were compared with competitive (c) inhibition ELISA. Microscopy being the least sensitive diagnostic test detected 12.0% of animals positive for A. marginale infection while nested-PCR detected 87.9% of these animals as positive for A. marginale infection. The recombinant MSP5 antigen showed positive reactivity in 170/190 nested-PCR confirmed positive animals (sensitivity 89.5%) with specificity of 77.0%. In comparison, the recombinant MSP2 antigen showed lesser sensitivity and specificity of 79.0% and 69.2%, respectively. The cELISA was more sensitive and specific than IgG-ELISA. However, molecular detection by msp5 nested-PCR was highly sensitive and reliable for detection of carrier cattle for Anaplasma infection. The study indicated that a large cattle population (87.9%) was carrier for A. marginale infection in this region of the country.
Assuntos
Anaplasma marginale , Anaplasmose , Doenças dos Bovinos , Anaplasma/genética , Anaplasma marginale/genética , Anaplasmose/diagnóstico , Anaplasmose/epidemiologia , Animais , Bovinos , Doenças dos Bovinos/diagnóstico , Doenças dos Bovinos/epidemiologia , Ensaio de Imunoadsorção Enzimática/veterináriaRESUMO
AIM: Serodiagnosis of Fasciola gigantica natural infection in buffaloes with recombinant cathepsin L1-D and native cathepsin-L protease antigens. METHODS: The recombinant cat L1-D antigen was expressed in prokaryotic expression system and native cathepsin-L proteases were purified by alcoholic fractionation from adult F. gigantica flukes. Buffaloes (n = 325) were screened for anti-Fasciola antibodies with the above antigens in immunoglobulin-G-enzyme linked immunosorbent assay (IgG-ELISA). RESULTS: The recombinant cat L1-D antigen showed positive reactivity with 101/122 necropsy positive animals but 21/122 necropsy confirmed positive animals were negative in this ELISA (sensitivity 82.8%). However, 30/203 (14.8%) necropsy negative animals for Fasciola were seropositive with specificity of 85.2%. With native cat-L protease, 104/122 necropsy confirmed positive animals were ELISA positive but 18/122 necropsy positive animals were seronegative, thereby depicting the sensitivity of 85.2%. But ELISA with this antigen showed 27/203 (13.3%) necropsy negative animals as positive (specificity 86.7%). CONCLUSIONS: Comparative evaluation of both the antigens showed that they are suitable for serodiagnosis of F. gigantica infection in buffalo herds.
Assuntos
Anticorpos Anti-Helmínticos/sangue , Búfalos/parasitologia , Catepsinas/imunologia , Cisteína Endopeptidases/imunologia , Fasciola/imunologia , Fasciolíase/veterinária , Proteínas de Helminto/imunologia , Animais , Western Blotting/veterinária , Catepsinas/genética , Catepsinas/metabolismo , Bovinos , Clonagem Molecular , Cisteína Endopeptidases/genética , Cisteína Endopeptidases/metabolismo , DNA Complementar/biossíntese , DNA Complementar/genética , Ensaio de Imunoadsorção Enzimática/veterinária , Fasciola/genética , Fasciola/isolamento & purificação , Fasciolíase/diagnóstico , Fasciolíase/parasitologia , Proteínas de Helminto/genética , Proteínas de Helminto/metabolismo , Fígado/parasitologia , Reação em Cadeia da Polimerase/veterinária , RNA de Helmintos/genética , RNA de Helmintos/isolamento & purificação , Coelhos , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/metabolismo , Sensibilidade e EspecificidadeRESUMO
Three recombinant proteins of Echinococcus granulosus including two antigen B sub-units EgAgB8/1 and EgAgB8/2 and Echinococcus protoscolex calcium binding protein 1 (EPC1) were expressed in prokaryotic expression vectors. The diagnostic potential of these three recombinant proteins was evaluated in the detection of cystic echinococcosis in buffaloes in IgG-ELISA. The EgAgB8/1 and EgAgB8/2 recombinant proteins reacted fairly with the hydatid infected buffaloes with sensitivity of 75.0% and 78.6%, respectively and specificity of 75.8% while EPC1 recombinant protein showed higher sensitivity (89.3%) but lower specificity (51.5%). Cross-reactivity of these three antigens was assayed with buffalo sera naturally infected with Explanatum explanatum, Paramphistomum epiclitum, Gastrothylax spp., Fasciola gigantica and Sarcocystis spp. EgAgB8/1 and EPC1 antigens cross-reacted with all these sera while EgAgB8/2 showed no cross-reaction with Sarcocystis spp. and reacted with some of the E. explanatum infected buffalo sera. This study explores the potential of three hydatid antigens viz. EgAgB8/1, EgAgB8/2 and EPC1 for their diagnostic potential in buffaloes positive for cystic echinococcosis.
