The cyanobacterial lectin, microvirin-N, enhances the specificity and sensitivity of lipoarabinomannan-based TB diagnostic tests.
The cyanobacterial lectin, microvirin-N, enhances the specificity and sensitivity of lipoarabinomannan-based TB diagnostic tests.
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DOI：
10.1039/d0an01725f
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发表时间：
2021-02-21
期刊：
影响因子：
--
通讯作者：
Wright DW
中科院分区：
文献类型：
--
作者：
van der Horst M ;Karamchand L ;Bauer WS ;Nel AJM ;Blackburn JM ;Wright DW
结核病（TB）是全球十大死亡原因之一，尽管可以治疗。除其他外，根除结核病需要在低资源环境中的护理点（POC）工作的具有高度特异性和敏感性的诊断测试。TB表面糖脂抗原，甘露糖封端的脂阿拉伯甘露聚糖（ManLAM）目前作为唯一的POC分子诊断生物标志物适用于低成本的免疫测定。在这里，我们证明了微病毒N（MVN）（一种14.3 kDa的蓝细菌凝集素）对H37 Rv TB ManLAM的高亲和力和卓越的特异性，并利用它开发了一种新型的珠上ELISA。MVN以亚皮摩尔结合亲和力结合ManLAM，但不结合由非致病性分枝杆菌表达的LAM的其他变体-当前市售抗LAM抗体不能实现的结合特异性和亲和力水平。随后使用MVN功能化磁珠开发了珠上ELISA，其允许从人尿液中特异性捕获ManLAM，检测限（LOD）为1.14 ng mL−1，并且当与PILAM（在非致病性分枝杆菌上发现的LAM变体）测试时没有交叉反应性。凝集素microvirin首次用于选择性结合结核病生物标志物ManLAM。
Tuberculosis (TB) is one of the top ten causes of death globally, despite being treatable. The eradication of TB disease requires, amongst others, diagnostic tests with high specificity and sensitivity that will work at the point of care (POC) in low-resource settings. The TB surface glycolipid antigen, mannose-capped lipoarabinomannan (ManLAM) currently serves as the only POC molecular diagnostic biomarker suitable for use in low cost immunoassays. Here, we demonstrate the high affinity and exceptional specificity of microvirin-N (MVN), a 14.3 kDa cyanobacterial lectin, toward H37Rv TB ManLAM and utilize it to develop a novel on-bead ELISA. MVN binds to ManLAM with sub-picomolar binding affinity, but does not bind to other variants of LAM expressed by non-pathogenic mycobacteria – a level of binding specificity and affinity that current commercially available anti-LAM antibodies cannot achieve. An on-bead ELISA was subsequently developed using MVN-functionalized magnetic beads which allows for the specific capture of ManLAM from human urine with a limit of detection (LOD) of 1.14 ng mL−1 and no cross-reactivity when tested with PILAM, a variant of LAM found on non-pathogenic mycobacteria. The lectin microvirin was used for the first time to selectively bind the tuberculosis biomarker ManLAM.
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影响因子：
11.3
作者：
Mishra AK;Driessen NN;Appelmelk BJ;Besra GS
通讯作者：
Besra GS
影响因子：
4.8
作者：
Huskens, Dana;Ferir, Geoffrey;Schols, Dominique
通讯作者：
Schols, Dominique
影响因子：
3.6
作者：
Kehr, JC;Zilliges, Y;Dittmann, E
通讯作者：
Dittmann, E
影响因子：
2.5
作者：
Lipman, NS;Jackson, LR;Weis-Garcia, F
通讯作者：
Weis-Garcia, F
DOI：
10.1016/j.trstmh.2005.04.014
发表时间：
2005-12-01
影响因子：
2.2
作者：
Boehme, C;Molokova, E;Hoelscher, M
通讯作者：
Hoelscher, M