Diagnostic probes for Bacillus anthracis spores selected from a landscape phage library
Diagnostic probes for Bacillus anthracis spores selected from a landscape phage library
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DOI：
10.1373/clinchem.2004.038018
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发表时间：
2004-10-01
影响因子：
9.3
通讯作者：
Petrenko, VA
中科院分区：
文献类型：
--
作者：
Brigati, J;Williams, DD;Petrenko, VA
y背景：最近炭疽杆菌孢子被用作生物武器，这突出表明需要有一个持续监测系统。目前的监测系统依赖于抗体衍生的探针，其不足以哈代以承受在极端条件下的长期使用。我们描述了新的，噬菌体衍生的探针，可以用作强大的替代品antibody.Methods：从一个景观噬菌体库与随机octapeptides上显示的所有拷贝的主要噬菌体外壳蛋白的噬菌体FD-四，我们选择的克隆，结合到孢子的B。炭疽菌（Sterne菌株）。ELISA、微量淘选和共沉淀试验用于评估这些噬菌体结合B的特异性和选择性。结果：筛选出的噬菌体克隆上的肽段可指导噬菌体与B结合。炭疽孢子大多数克隆表现出很少或没有绑定到孢子的远亲芽孢杆菌属物种，但观察到一些绑定与孢子的密切相关的物种。我们最特异的孢子结合噬菌体展示肽EPRLSPHS（每个噬菌体几千肽），并且与B的孢子的结合好3.5至70倍。结论：筛选出的噬菌体探针优先与B结合。炭疽菌Sterne孢子与其他芽孢杆菌属物种相比。这些噬菌体有可能进一步发展成为高度特异性和稳健的探针，适用于连续监测设备和生物吸附剂的长期使用。(C)2004年美国临床化学协会。
yBackground: Recent use of Bacillus anthracis spores as a bioweapon has highlighted the need for a continuous monitoring system. Current monitoring systems rely on antibody-derived probes, which are not hardy enough to withstand long-term use under extreme conditions. We describe new, phage-derived probes that can be used as robust substitutes for antibodies.Methods: From a landscape phage library with random octapeptides displayed on all copies of the major phage coat protein of the phage fd-tet, we selected clones that bound to spores of B. anthracis (Sterne strain). ELISA, micropanning, and coprecipitation assays were used to evaluate the specificity and selectivity with which these phage bound to B. anthracis spores.Results: Peptides on the selected clones directed binding of the phage to B. anthracis spores. Most clones exhibited little or no binding to spores of distantly related-Bacillus species, but some binding was observed with spores of closely related species. Our most specific spore-binding phage displayed a peptide EPRLSPHS (several thousand peptides per phage) and bound 3.5- to 70-fold better to spores of B. anthracis Sterne than to spores of other Bacillus species.Conclusions: The selected phage probes bound preferentially to B. anthracis Sterne spores compared with other Bacillus species. These phage could possibly be further developed into highly specific and robust probes suitable for long-term use in continuous monitoring devices and biosorbents. (C) 2004 American Association for Clinical Chemistry.