Tabaqchali S, Silman R, Holland D
Department of Medical Microbiology, St Bartholomew's Hospital Medical College, London.
J Clin Pathol. 1987 Sep;40(9):1070-87. doi: 10.1136/jcp.40.9.1070.
A new rapid automated method for the identification and classification of microorganisms is described. It is based on the incorporation of 35S-methionine into cellular proteins and subsequent separation of the radiolabelled proteins by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The protein patterns produced were species specific and reproducible, permitting discrimination between the species. A large number of Gram negative and Gram positive aerobic and anaerobic organisms were successfully tested. Furthermore, there were sufficient differences within species between the protein profiles to permit subdivision of the species. New typing schemes for Clostridium difficile, coagulase negative staphylococci, and Staphylococcus aureus, including the methicillin resistant strains, could thus be introduced; this has provided the basis for useful epidemiological studies. To standardise and automate the procedure an automated electrophoresis system and a two dimensional scanner were developed to scan the dried gels directly. The scanner is operated by a computer which also stores and analyses the scan data. Specific histograms are produced for each bacterial species. Pattern recognition software is used to construct databases and to compare data obtained from different gels: in this way duplicate "unknowns" can be identified. Specific small areas showing differences between various histograms can also be isolated and expanded to maximise the differences, thus providing differentiation between closely related bacterial species and the identification of differences within the species to provide new typing schemes. This system should be widely applied in clinical microbiology laboratories in the near future.
本文描述了一种用于微生物鉴定和分类的新型快速自动化方法。该方法基于将³⁵S-甲硫氨酸掺入细胞蛋白质中,随后通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)分离放射性标记的蛋白质。所产生的蛋白质图谱具有物种特异性且可重复,能够区分不同物种。大量革兰氏阴性和革兰氏阳性需氧菌和厌氧菌均已成功测试。此外,同一物种内的蛋白质谱存在足够差异,可对物种进行细分。由此可以引入针对艰难梭菌、凝固酶阴性葡萄球菌和金黄色葡萄球菌(包括耐甲氧西林菌株)的新分型方案;这为有用的流行病学研究提供了基础。为了使该程序标准化和自动化,开发了一种自动电泳系统和二维扫描仪,用于直接扫描干燥的凝胶。扫描仪由计算机操作,计算机还存储和分析扫描数据。为每个细菌物种生成特定的直方图。模式识别软件用于构建数据库并比较从不同凝胶获得的数据:通过这种方式可以识别重复的“未知物”。还可以分离并放大显示不同直方图之间差异的特定小区域,以最大化差异,从而区分密切相关的细菌物种并识别物种内的差异,以提供新的分型方案。该系统在不久的将来应会在临床微生物学实验室中得到广泛应用。