Pietrzak J, Muehlestein S, Gasser M
Institut für Medizinische Mikrobiologie der Universität Basel.
Zentralbl Bakteriol. 1990 Oct;274(1):61-9. doi: 10.1016/s0934-8840(11)80975-8.
In this paper we describe a new assay for diphtheria toxin in bacterial cultures, based on a sandwich-dot immunobinding method. This method uses horse polyclonal diphtheria antitoxin as the coating antibody and mouse monoclonal diphtheria antitoxin as the detecting antibody. The sensitivity of this new method is within a range of 5-10 ng toxin per ml in Corynebacterium diphtheriae cultures. We did not observe any false-positive reactions. As a solid phase we used nitrocellulose disks which bind protein more strongly than the commonly used plastic solid phase. Results with strong toxin producers of C. diphtheriae can be obtained as soon as 11 h after starting a bacterial test culture. To detect weak toxin producers, the cultivation period must be extended to about 18 h and the results can be obtained after about 24 h. The method presented in this paper is simpler, faster and more reliable than currently used methods.
在本文中，我们描述了一种基于夹心斑点免疫结合法检测细菌培养物中白喉毒素的新方法。该方法使用马多克隆白喉抗毒素作为包被抗体，小鼠单克隆白喉抗毒素作为检测抗体。这种新方法的灵敏度在每毫升白喉棒状杆菌培养物中5 - 10纳克毒素的范围内。我们未观察到任何假阳性反应。作为固相，我们使用了硝酸纤维素圆盘，其比常用的塑料固相更牢固地结合蛋白质。对于白喉棒状杆菌的强毒素产生菌，在开始细菌测试培养后11小时即可获得结果。为了检测弱毒素产生菌，培养期必须延长至约18小时，约24小时后可获得结果。本文介绍的方法比目前使用的方法更简单、更快且更可靠。
