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Registro Completo |
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Biblioteca(s): |
Embrapa Pecuária Sudeste. |
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Data corrente: |
27/05/2024 |
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Data da última atualização: |
22/08/2024 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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Autoria: |
GIGLIOTI, R.; VERCESI FILHO, A. E.; DOMINGOS, A. G.; SILVA, S. S. DA; CUNHA, R. C.; IBELLI, A. M. G.; OKINO, C. H.; OLIVEIRA, M. C. de S. |
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Afiliação: |
RODRIGO GIGLIOTI, INSTITUTO DE ZOOTECNIA; ANIBAL EUGÊNIO VERCESI FILHO, INSTITUTO DE ZOOTECNIA; ANA GONÇALVES DOMINGOS, UNIVERSIDADE NOVA DE LISBOA; SÉRGIO SILVA DA SILVA, C. R. O. ANIMAL SCIENCE; RODRIGO CASQUERO CUNHA, UNIVERSIDADE FEDERAL DE PELOTAS; ADRIANA MERCIA GUARATINI IBELLI, CPPSE; CINTIA HIROMI OKINO, CPPSE; MARCIA CRISTINA DE SENA OLIVEIRA, CPPSE. |
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Título: |
Detection and quantification of Babesia bovis and Babesia bigemina using different target genes. |
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Ano de publicação: |
2024 |
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Fonte/Imprenta: |
Research in Veterinary Science, v. 168, 2024, 105122. |
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Páginas: |
7 p. |
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DOI: |
10.1016/j.rvsc.2023.105122 |
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Idioma: |
Inglês |
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Conteúdo: |
Molecular assays have been widely used for the detection and quantification of bovine babesiosis due to their high sensitivity and specificity. However, variations in the sensitivity of pathogen detection may occur depending on the selected target gene. Thus, this study aimed to compare the detection sensitivity (DS) of Babesia bovis and B. bigemina infection levels in artificially and naturally infected cattle using quantitative PCR (qPCR) and six target genes. For B. bovis, the merozoite surface antigen genes 2b and 2c (msa-2b and msa-2c), and the mitochondrial cytochrome b gene (cybmt) were used. For B. bigemina, the genes encoding the proteins associated with rhoptry 1c (rap-1c), rap-1a, and cybmt were used. Six bovines, free of babesiosis, were artificially infected with 1 × 10 8 red blood cells infected (iRBC) with B. bovis (n = 3) or 1 × 10 6 B. bigemina iRBC (n = 3). The animals were evaluated daily until parasitemia was confirmed (≥ 2.0%). The quantity of iRBC present in each animal was determined by examining blood smears. Blood samples were then subjected to DNA extraction, serial dilution, and qPCR analysis to determine the DS of each target gene. In addition, 30 calves naturally infected by Babesia spp. were also evaluated using the same six target genes. Regarding the artificial infection, B. bovis cybmt showed 25-fold higher sensitivity than the msa-2b and msa-2c genes, while the B. bigemina cybmt exhibited 5-fold and 25-fold higher sensitivity than the rap-1a and rap-1c genes, respectively. The rap-1a gene was found to be 5 times more sensitive than the rap-1c gene, while the B. bovis msa-2b and msa-2c genes exhibited similar DS. The positive frequencies of naturally infected calves for the target cybmt, msa-2b, and msa-2c genes (B. bovis) were: 100%, 33.3% and 50%, while cybmt, rap-1a, and rap-1c genes (B. bigemina) were 90%, 83.3%, and 63.3%, respectively. This study may contribute to the selection of suitable genes for molecular monitoring of bovine babesiosis. Mitochondrial genes could be considered as an alternative to improve the sensitivity of B. bovis and B. bigemina detection using qPCR. MenosMolecular assays have been widely used for the detection and quantification of bovine babesiosis due to their high sensitivity and specificity. However, variations in the sensitivity of pathogen detection may occur depending on the selected target gene. Thus, this study aimed to compare the detection sensitivity (DS) of Babesia bovis and B. bigemina infection levels in artificially and naturally infected cattle using quantitative PCR (qPCR) and six target genes. For B. bovis, the merozoite surface antigen genes 2b and 2c (msa-2b and msa-2c), and the mitochondrial cytochrome b gene (cybmt) were used. For B. bigemina, the genes encoding the proteins associated with rhoptry 1c (rap-1c), rap-1a, and cybmt were used. Six bovines, free of babesiosis, were artificially infected with 1 × 10 8 red blood cells infected (iRBC) with B. bovis (n = 3) or 1 × 10 6 B. bigemina iRBC (n = 3). The animals were evaluated daily until parasitemia was confirmed (≥ 2.0%). The quantity of iRBC present in each animal was determined by examining blood smears. Blood samples were then subjected to DNA extraction, serial dilution, and qPCR analysis to determine the DS of each target gene. In addition, 30 calves naturally infected by Babesia spp. were also evaluated using the same six target genes. Regarding the artificial infection, B. bovis cybmt showed 25-fold higher sensitivity than the msa-2b and msa-2c genes, while the B. bigemina cybmt exhibited 5-fold and 25-fold higher sensitivity than the rap-1a... Mostrar Tudo |
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Palavras-Chave: |
QPCR; Quantification; Sensitivity. |
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Thesaurus Nal: |
Babesiosis; Detection. |
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Categoria do assunto: |
H Saúde e Patologia |
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Marc: |
LEADER 02980naa a2200289 a 4500
001 2164535
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008 2024 bl uuuu u00u1 u #d
024 7 $a10.1016/j.rvsc.2023.105122$2DOI
100 1 $aGIGLIOTI, R.
245 $aDetection and quantification of Babesia bovis and Babesia bigemina using different target genes.$h[electronic resource]
260 $c2024
300 $a7 p.
520 $aMolecular assays have been widely used for the detection and quantification of bovine babesiosis due to their high sensitivity and specificity. However, variations in the sensitivity of pathogen detection may occur depending on the selected target gene. Thus, this study aimed to compare the detection sensitivity (DS) of Babesia bovis and B. bigemina infection levels in artificially and naturally infected cattle using quantitative PCR (qPCR) and six target genes. For B. bovis, the merozoite surface antigen genes 2b and 2c (msa-2b and msa-2c), and the mitochondrial cytochrome b gene (cybmt) were used. For B. bigemina, the genes encoding the proteins associated with rhoptry 1c (rap-1c), rap-1a, and cybmt were used. Six bovines, free of babesiosis, were artificially infected with 1 × 10 8 red blood cells infected (iRBC) with B. bovis (n = 3) or 1 × 10 6 B. bigemina iRBC (n = 3). The animals were evaluated daily until parasitemia was confirmed (≥ 2.0%). The quantity of iRBC present in each animal was determined by examining blood smears. Blood samples were then subjected to DNA extraction, serial dilution, and qPCR analysis to determine the DS of each target gene. In addition, 30 calves naturally infected by Babesia spp. were also evaluated using the same six target genes. Regarding the artificial infection, B. bovis cybmt showed 25-fold higher sensitivity than the msa-2b and msa-2c genes, while the B. bigemina cybmt exhibited 5-fold and 25-fold higher sensitivity than the rap-1a and rap-1c genes, respectively. The rap-1a gene was found to be 5 times more sensitive than the rap-1c gene, while the B. bovis msa-2b and msa-2c genes exhibited similar DS. The positive frequencies of naturally infected calves for the target cybmt, msa-2b, and msa-2c genes (B. bovis) were: 100%, 33.3% and 50%, while cybmt, rap-1a, and rap-1c genes (B. bigemina) were 90%, 83.3%, and 63.3%, respectively. This study may contribute to the selection of suitable genes for molecular monitoring of bovine babesiosis. Mitochondrial genes could be considered as an alternative to improve the sensitivity of B. bovis and B. bigemina detection using qPCR.
650 $aBabesiosis
650 $aDetection
653 $aQPCR
653 $aQuantification
653 $aSensitivity
700 1 $aVERCESI FILHO, A. E.
700 1 $aDOMINGOS, A. G.
700 1 $aSILVA, S. S. DA
700 1 $aCUNHA, R. C.
700 1 $aIBELLI, A. M. G.
700 1 $aOKINO, C. H.
700 1 $aOLIVEIRA, M. C. de S.
773 $tResearch in Veterinary Science$gv. 168, 2024, 105122.
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Registro original: |
Embrapa Pecuária Sudeste (CPPSE) |
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