Fujinami Yoshihito, Hirai Yoshikazu, Sakai Ikuko, Yoshino Mineo, Yasuda Jiro
Department of First Forensic Science, National Research Institute of Police Science, Kashiva, Chiba, Japan.
Microbiol Immunol. 2007;51(2):163-9. doi: 10.1111/j.1348-0421.2007.tb03894.x.
Detection of biological weapons is a primary concern in force protection, treaty verification, and safeguarding civilian populations against domestic terrorism. One great concern is the detection of Bacillus anthracis, the causative agent of anthrax. Therefore, there is a pressing need to develop novel methods for rapid, simple, and precise detection of B. anthracis. Here, we report that the C-terminal region of gamma-phage lysin protein (PlyG) binds specifically to the cell wall of B. anthracis and the recombinant protein corresponding to this region (positions, 156-233), PlyGB, is available as a bioprobe for detection of B. anthracis. Our detection method, based on a membrane direct blot assay using recombinant PlyGB, was more rapid and sensitive than the gamma-phage test and was simpler and more inexpensive than genetic methods such as PCR, or immunological methods using specific antibodies. Furthermore, its specificity was comparable to the gamma-phage test. PlyGB is applicable in conventional methods instead of antibodies and could be a potent tool for detection of B. anthracis.
生物武器的检测是部队防护、条约核查以及保护平民免受国内恐怖主义威胁的首要关注点。一个重大担忧是炭疽杆菌(炭疽病的病原体)的检测。因此,迫切需要开发用于快速、简便且精确检测炭疽杆菌的新方法。在此,我们报告γ-噬菌体溶菌蛋白(PlyG)的C末端区域特异性结合炭疽杆菌的细胞壁,并且对应于该区域(第156 - 233位)的重组蛋白PlyGB可作为检测炭疽杆菌的生物探针。我们基于使用重组PlyGB的膜直接印迹分析的检测方法比γ-噬菌体检测更快、更灵敏,并且比诸如PCR等基因方法或使用特异性抗体的免疫方法更简单、更便宜。此外,其特异性与γ-噬菌体检测相当。PlyGB可替代抗体应用于传统方法,并且可能成为检测炭疽杆菌的有力工具。