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

被引：13
作者：
Giglioti, Rodrigo [1 ,2 ,3 ]
Bassetto, Cesar Cristiano [1 ,2 ]
Okino, Cintia Hiromi [2 ]
de Oliveira, Henrique Nunes [1 ]
de Sena Oliveira, Marcia Cristina [2 ]
机构：
[1] Univ Estadual Paulista, Fac Ciencias Agr & Vet, Via Acesso Prof Paulo Donato Castellane, BR-14884900 Jaboticabal, SP, Brazil
[2] Embrapa Pecuaria Sudeste, Rodovia Washington Luiz,Km 234,CP 339, BR-13560970 Sao Carlos, SP, Brazil
[3] Ctr Pesquisa Genet & Reprod Anim, Inst Zootecnia, Rua Heitor Penteado 56, BR-13380011 Nova Odessa, SP, Brazil
来源：
EXPERIMENTAL AND APPLIED ACAROLOGY | 2019年 / 77卷 / 01期
基金：
巴西圣保罗研究基金会;
关键词：
LAMP; qPCR; Anaplasma marginale; Detection; Sensibility; Specificity; RAPID DETECTION; CATTLE;
D O I：
10.1007/s10493-018-0327-y
中图分类号：
Q96 [昆虫学];
学科分类号：
摘要：
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.
引用
收藏
页码：65 / 72
页数：8
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