Development of a loop-mediated isothermal amplification (LAMP) assay for the detection of Anaplasma marginale

Autor(es)

Giglioti, Rodrigo ^{[1, 2, 3]} ; Bassetto, Cesar Cristiano ^{[1, 2]} ; Okino, Cintia Hiromi ^[1] ; de Oliveira, Henrique Nunes ^[2] ; de Sena Oliveira, Marcia Cristina ^[1]

Afiliação do(s) autor(es)

^[1] Embrapa Pecuaria Sudeste, Rodovia Washington Luiz, Km 234, CP 339, BR-13560970 Sao Carlos, SP - Brazil

^[2] Univ Estadual Paulista, Fac Ciencias Agr & Vet, Via Acesso Prof Paulo Donato Castellane, BR-14884900 Jaboticabal, SP - Brazil

^[3] Ctr Pesquisa Genet & Reprod Anim, Inst Zootecnia, Rua Heitor Penteado 56, BR-13380011 Nova Odessa, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: Experimental and Applied Acarology; v. 77, n. 1, p. 65-72, JAN 2019.
• Citações Web of Science: 1

Resumo

Parasitemia generated by Anaplasma marginale causes significant losses in the cattle industry. A major constraint to the effective control and management of anaplasmosis in livestock is the lack of a rapid and reliable diagnostic test to identify the parasite and allow for immediate therapy. In the present study, we developed a novel DNA-based assay for the detection of A. marginale in bovine blood samples, using loop-mediated isothermal amplification (LAMP). DNA from six cattle and hemoparasite samples (Babesia bovis, Babesia bigemina, Anaplasma centrale and A. marginale) were tested for specificity, sensitivity and cross-reactions. The developed LAMP procedures were also confirmed and compared with the qPCR method. The same gene sequence (major surface protein 1b, msp1b) of A. marginale was used to design a set of primers for the LAMP and qPCR assays. The results showed that LAMP is specific, as no positive signal was observed for the other tested hemoparasites. However, the sensitivity of the qPCR assay was ten times higher than LAMP. Our findings indicate that this LAMP method has a good sensitivity and high specificity for the detection of A. marginale and may have a potential application in the detection and differentiation of bovine anaplasmosis. (AU)

• Processo FAPESP: 16/07216-7 - Estudo genômico e caracterização imunológica de bovinos resistentes à babesiose bovina e análise da diversidade genética de Babesia bovis e Babesia bigemina
• Beneficiário: Henrique Nunes de Oliveira
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 17/11297-5 - CARACTERIZAÇÃO IMUNOLÓGICA DE BOVINOS RESISTENTES À BABESIOSE BOVINA E ANÁLISE DA DIVERSIDADE GENÉTICA DE Babesia bovis E Babesia bigemina
• Beneficiário: Rodrigo Giglioti
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado