Detection and quantification of Babesia bovis and Babesia bigemina using different target genes.

Autor: Giglioti R; Instituto de Zootecnia, Rua Heitor Penteado, n. 56, Nova Odessa, São Paulo 13380-011, Brazil. Electronic address: rodrigo.giglioti@sp.gov.br., Filho AEV; Instituto de Zootecnia, Rua Heitor Penteado, n. 56, Nova Odessa, São Paulo 13380-011, Brazil., Domingos AG; Instituto de Higiene e Medicina Tropical, Universidade Nova de Lisboa, Portugal., da Silva SS; C.R.O. Animal Science, Estrada Colônia São Domingos, Colônia, Turuçú, Rio Grande do Sul, Brazil., Cunha RC; Laboratório de Biologia Molecular Veterinária, Faculdade de Veterinária, Universidade Federal de Pelotas, Capão do Leão, Rio Grande do Sul, Brazil., Ibelli AMG; Embrapa Pecuária Sudeste, São Carlos, São Paulo, Brazil., Okino CH; Embrapa Pecuária Sudeste, São Carlos, São Paulo, Brazil., de Sena Oliveira MC; Embrapa Pecuária Sudeste, São Carlos, São Paulo, Brazil.
Jazyk: angličtina
Zdroj: Research in veterinary science [Res Vet Sci] 2024 Mar; Vol. 168, pp. 105122. Date of Electronic Publication: 2023 Dec 26.
DOI: 10.1016/j.rvsc.2023.105122
Abstrakt: 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.
Competing Interests: Declaration of Competing Interest The authors declare that there is no competing interest or conflicts of interest.
(Copyright © 2023. Published by Elsevier Ltd.)
Databáze: MEDLINE
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