Proteomic Identification of Immunodiagnostic Antigens for Trypanosoma vivax Infections in Cattle and Generation of a Proof-of-Concept Lateral Flow Test Diagnostic Device.
Proteomic Identification of Immunodiagnostic Antigens for Trypanosoma vivax Infections in Cattle and Generation of a Proof-of-Concept Lateral Flow Test Diagnostic Device.
复制标题
DOI：
10.1371/journal.pntd.0004977
复制
发表时间：
2016-09
影响因子：
3.8
通讯作者：
Ferguson MA
中科院分区：
文献类型：
--
作者：
Fleming JR;Sastry L;Wall SJ;Sullivan L;Ferguson MA
间日锥虫是非洲动物锥虫病的病原体之一，在撒哈拉以南非洲流行，主要通过采采蝇媒介的叮咬传播。这种寄生虫也可以通过机械传播，这使得它传播到了南美洲。这种寄生虫的诊断是有限的，在农场环境中，诊断主要是基于寄生虫的。我们开始使用蛋白质组学方法鉴定候选诊断抗原，以开发成易于使用的笔侧侧流测试装置。选择不变表面糖蛋白家族的两个相关成员TvY486_0045500和TvY486_0019690。这些抗原的片段，缺乏N-末端信号肽和C-末端跨膜结构域，在E.杆菌两者都被开发成ELISA测试，其中之一TvY486_0045500被开发成侧向流测试原型。所有试验均采用113份随机血清样本进行盲法评价，这些样本取自37头感染T.间日疟原虫或T.刚果语。TvY486_0045500和TvY486_0019690 ELISA试验对T的灵敏度和特异性值相同。间日疟感染率94.5%（95% CI，86.5%至98.5%）和88.0%（95%CI，75.7%至95.5%），而TvY486_0045500侧流试验原型的灵敏度和特异性为92.0%（95% CI，83.4%至97.0%）和89.8%（95% CI，77.8%至96.6%）。这些数据表明，重组TvY486_0045500显示出开发用于诊断T.非洲锥虫病。非洲动物锥虫病给撒哈拉以南非洲的农业发展带来了严重的问题，并导致了巨大的经济损失。其中一种主要的寄生虫是间日锥虫。目前的诊断方法要么是以生物学为基础的，要么成本太高，技术要求太高，无法在流行地区使用。在这里，我们确定了T。间日疟原虫蛋白被感染的牛血清选择性识别，并将两种相关蛋白开发成ELISA测试，其中一种开发成侧向流测试原型。当针对随机小牛血清进行检测时，所有三种检测均表现良好，表明笔侧T的开发潜力良好。用于流行地区的间日动物非洲锥虫病诊断装置。
Trypanosoma vivax is one of the causative agents of Animal African Trypanosomosis in cattle, which is endemic in sub-Saharan Africa and transmitted primarily by the bite of the tsetse fly vector. The parasite can also be mechanically transmitted, and this has allowed its spread to South America. Diagnostics are limited for this parasite and in farm settings diagnosis is mainly symptom-based. We set out to identify, using a proteomic approach, candidate diagnostic antigens to develop into an easy to use pen-side lateral flow test device. Two related members the invariant surface glycoprotein family, TvY486_0045500 and TvY486_0019690, were selected. Segments of these antigens, lacking N-terminal signal peptides and C-terminal transmembrane domains, were expressed in E. coli. Both were developed into ELISA tests and one of them, TvY486_0045500, was developed into a lateral flow test prototype. The tests were all evaluated blind with 113 randomised serum samples, taken from 37 calves before and after infection with T. vivax or T. congolense. The TvY486_0045500 and TvY486_0019690 ELISA tests gave identical sensitivity and specificity values for T. vivax infection of 94.5% (95% CI, 86.5% to 98.5%) and 88.0% (95% CI, 75.7% to 95.5%), respectively, and the TvY486_0045500 lateral flow test prototype a sensitivity and specificity of 92.0% (95% CI, 83.4% to 97.0%) and 89.8% (95% CI, 77.8% to 96.6%), respectively. These data suggest that recombinant TvY486_0045500 shows promise for the development of a pen-side lateral flow test for the diagnosis of T. vivax animal African trypanosomosis. African Animal Trypanosomosis presents a significant problem for agricultural development in sub-Saharan Africa and leads to large economic losses. One of the main parasites responsible is Trypanosoma vivax. Current diagnostic methods are either symptom-based or too costly and technologically demanding for use in endemic regions. Here, we identified T. vivax proteins selectively recognized by infected cattle sera and developed two related proteins into ELISA tests and one of these into a lateral flow test prototype. All three tests performed well when tested against randomised calf sera, suggesting good potential for the development of a pen-side T. vivax animal African trypanosomosis diagnostic device for use in endemic regions.
登录
查看更多内容
影响因子：
2.6
作者：
IJAGBONE, IF;STAAK, C;REINHARD, R
通讯作者：
REINHARD, R
影响因子：
3.8
作者：
Sullivan, Lauren;Fleming, Jennifer;Ferguson, Michael A. J.
通讯作者：
Ferguson, Michael A. J.
影响因子：
--
作者：
Tran, Thao;Claes, Filip;Buescher, Philippe
通讯作者：
Buescher, Philippe
影响因子：
2.6
作者：
Uzcanga, Graciela L.;Perez-Rojas, Yenis;Bubis, Jose
通讯作者：
Bubis, Jose
影响因子：
3.8
作者：
Sternberg, Jeremy M.;Gierlinski, Marek;Ndung'u, Joseph M.
通讯作者：
Ndung'u, Joseph M.