Clonagem, expressão, caracterizaÇão molecular da proteína de superfície MSP5 da amostra PR1 de Anaplasma marginale e sua aplicaÇão em um teste de ELISA por competiÇão

Marana, Elizabete Regina Marangoni; Kano, Flora Satiko; Vicentini, Josy Campanhã; Spurio, Rafael Sanches; Ribeiro, M. F. B.; Coelho, Adriana Letícia M; Vidotto, Marilda Carlos; Vidotto, Odilon

doi:10.4322/rbpv.01802002

Cited by 2 publications

(2 citation statements)

References 20 publications

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“…To determine sensitivity and specificity of IFAT, it was compared to the ELISA test, which was considered to be the gold standard because it presents high specificity and produces few false positive results (MARANA et al, 2009;BRAZ et al, 2000).…”

Section: Resultsmentioning

confidence: 99%

Transplacental transmission of Anaplasma marginale in beef cattle chronically infected in southern Brazil

Grau

Filho

Pappen

et al. 2013

Rev. Bras. Parasitol. Vet.

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In this study, we have investigated the incidence of transplacental transmission of Anaplasma marginale in chronically infected cows with no history of acute anaplasmosis during gestation. In addition, we evaluated various techniques for detection of transplacental transmission of A. marginale. Blood samples were collected from 30 cows at three different periods: at the time of artificial insemination, at gestational diagnosis, and after calving. Also, blood was collected from the newborn calves, including one sample before colostrum intake, and another three days after birth. A. marginale-specific antibodies were detected in 100% of the cows with an indirect fluorescent antibody test (IFAT), and in 97% of them, using an indirect enzyme-linked immunosorbent assay (ELISA). Also, we observed that all of the three-day-old newborn calves were seropositive by IFAT. According to polymerase chain reaction, 63.3% of the cows were carriers of A. marginale, as well as 6.7% of the newborn calves. This represented a transplacental transmission rate of 10.5%. Furthermore, a correlation of 93.3% was observed between the two serodiagnostic techniques, demonstrating that both ELISA and IFAT can be used in epidemiological surveys of A. marginale. These results confirm the occurrence of transplacental transmission of A. marginale in chronically infected cows and suggest the importance of this transmission route in areas of enzootic instability.Keywords: Anaplasma marginale, transplacental transmission, enzootic instability. ResumoO objetivo do estudo foi avaliar a transmissão transplacentária de A. marginale em bovinos de corte cronicamente infectados utilizando diferentes testes de diagnóstico. Amostras de sangue foram colhidas de trinta matrizes durante o período de inseminação artificial, no diagnóstico de gestação e após o parto. Foi colhido sangue dos bezerros antes da ingestão do colostro e três dias após o nascimento. Anticorpos foram detectados em 100% das matrizes pela Reação de Imunofluorescência Indireta (RIFI) e 97% pelo Ensaio de Imunoadsorção Enzimático Indireto (ELISA). Aos três dias de idade 100% dos bezerros estavam soropositivos na RIFI. Conforme os resultados da PCR, 63.3% das matrizes e 6.7% dos bezerros eram portadores do agente, representando taxa de transmissão transplacentária de 10.5%. Correlação de 93.3% foi observada entre os resultados das técnicas sorológicas, demonstrando que tanto ELISA quanto RIFI podem ser utilizadas em levantamentos epidemiológicos do agente. Os resultados confirmaram a transmissão transplacentária do agente em vacas cronicamente infectadas. Isto demonstra a importância desta via de transmissão do A. marginale em áreas de instabilidade enzoótica.Palavras-chave: Anaplasma marginale, transmissão transplacentária, instabilidade enzoótica.

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Section: Resultsmentioning

confidence: 99%

Transplacental transmission of Anaplasma marginale in beef cattle chronically infected in southern Brazil

Grau

Filho

Pappen

et al. 2013

Rev. Bras. Parasitol. Vet.

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“…Escherichia coli BL21 Star (DE3) One Shot cells (Invitrogen Life Technologies, Sao Paulo, BRA) were transformed with the recombinant plasmids pET102-MSP1a, pET102-MSP1b, pET102-MSP4 and pET102-MSP5 to produce recombinant MSP1a and MSP1b (TAMEKUNI et al, 2009), MSP4 (KAWASAKI et al, 2007a and MSP5 (MARANA et al, 2009) as a fusion product with 6His. The transformants were grown in Luria Bertani (LB) medium to an optical density (OD) of OD 500nm 0.7, and Isopropilthio-D-Galactoside (IPTG) was added to a final concentration of 1 mM.…”

Section: Expression Of Msp Genes and Purification Of Recombinant Mspsmentioning

confidence: 99%

Immune response of calves inoculated with proteins ofAnaplasma marginale bound to an immunostimulant complex

Gasparini

Vieira

Nascimento

et al. 2013

Rev. Bras. Parasitol. Vet.

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Despite our current knowledge of the immunology, pathology, and genetics of Anaplasma marginale, prevention in cattle is currently based on old standbys, including live attenuated vaccines, antibiotic treatment, and maintaining enzootic stability in cattle herds. In the present study, we evaluated the use of an immunostimulant complex (ISCOMATRIX) adjuvant, associated with a pool of recombinant major surface proteins (rMSP1a, rMSP1b, rMSP4 and rMSP5) to improve the humoral immune response triggered in calves mainly by IgG2. Ten calves were divided in three groups: 4 calves were inoculated with the ISCOMATRIX/rMSPs (G1); 2 calves were inoculated with ISCOMATRIX adjuvant (G2); and 4 calves received saline (G3). Three inoculations were administered at 21-day intervals. In G1, the calves showed significant increases in total IgG, IgG1 and IgG2 levels 21 days after the second inoculation, compared to the control group (p < 0.05), and G1 calves remained above the cut-off value 28 days after the third inoculation (p < 0.05). The post-immunized sera from calves in G1 reacted specifically for each of the rMSPs used. In conclusion, the ISCOMATRIX/rMSPs induced antigen-specific seroconversion in calves. Therefore, additional testing to explore the protection induced by rMSPs, both alone and in conjunction with proteins previously identified as subdominant epitopes, is warranted.

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Clonagem, expressão, caracterizaÇão molecular da proteína de superfície MSP5 da amostra PR1 de Anaplasma marginale e sua aplicaÇão em um teste de ELISA por competiÇão

Cited by 2 publications

References 20 publications

Transplacental transmission of Anaplasma marginale in beef cattle chronically infected in southern Brazil

Transplacental transmission of Anaplasma marginale in beef cattle chronically infected in southern Brazil

Immune response of calves inoculated with proteins ofAnaplasma marginale bound to an immunostimulant complex

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