Development of a Nanobody-based lateral flow assay to detect active Trypanosoma congolense infections.

Development of a Nanobody-based lateral flow assay to detect active Trypanosoma congolense infections.
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DOI:
10.1038/s41598-018-26732-7
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发表时间:
2018-06-13
期刊:
影响因子:
4.6
通讯作者:
Magez S
Magez S
中科院分区:
综合性期刊3区
文献类型:
--
作者:
Pinto Torres JE;Goossens J;Ding J;Li Z;Lu S;Vertommen D;Naniima P;Chen R;Muyldermans S;Sterckx YG;Magez S

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非洲动物锥虫病(AAT)是一种影响家畜的疾病,由锥虫属寄生虫(主要是间日弓形虫和弓形虫)引起。AAT在撒哈拉以南非洲很普遍,它继续给那里造成沉重的社会经济负担,因为它使可持续牲畜饲养的发展非常艰难。在开始药物治疗之前主动发现病例和识别受感染的动物,需要有敏感和具体的诊断试验。本文介绍了利用纳米抗体(NBS)检测弓形虫的两种异源夹心法(EL ISA和LFA)的发展。用来自两个弓形虫菌株的分泌组混合物免疫羊驼,鉴定出一对针对糖酵解酶丙酮酸激酶的NB对(Nb44/Nb42)。我们证明,NB44/NB42酶联免疫吸附试验和LFA可用于检测实验感染的小鼠和牛的血浆样本中的寄生虫血症,此外,它们还可以作为“治愈测试”工具。综上所述,本文的发现展示了第一个基于NB的抗原检测LFA的发展和评估,以识别活动的弓形虫感染。
Animal African trypanosomosis (AAT), a disease affecting livestock, is caused by parasites of the Trypanosoma genus (mainly T. vivax and T. congolense). AAT is widespread in Sub-Saharan Africa, where it continues to impose a heavy socio-economic burden as it renders development of sustainable livestock rearing very strenuous. Active case-finding and the identification of infected animals prior to initiation of drug treatment requires the availability of sensitive and specific diagnostic tests. In this paper, we describe the development of two heterologous sandwich assay formats (ELISA and LFA) for T. congolense detection through the use of Nanobodies (Nbs). The immunisation of an alpaca with a secretome mix from two T. congolense strains resulted in the identification of a Nb pair (Nb44/Nb42) that specifically targets the glycolytic enzyme pyruvate kinase. We demonstrate that the Nb44/Nb42 ELISA and LFA can be employed to detect parasitaemia in plasma samples from experimentally infected mice and cattle and, additionally, that they can serve as ‘test-of-cure’ tools. Altogether, the findings in this paper present the development and evaluation of the first Nb-based antigen detection LFA to identify active T. congolense infections.
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