Brooks K, Sodeman T
Appl Microbiol. 1974 Jun;27(6):1023-6. doi: 10.1128/am.27.6.1023-1026.1974.
Deoxyribonucleic acid (DNA) from 250 strains of aerobic, nonfermentative, gram-negative coccobacilli and rods were tested for the ability to transform a stable competent auxotroph of Acinetobacter (strain trp E 27) to prototrophy by using the method established by Juni. Several modifications of Juni's original procedure were made to adapt it for use in a clinical diagnostic laboratory. These modifications were directed primarily towards shortening the procedure to allow completion in a time framework consistent with current procedures. The modifications included changes in sterilization temperature, incubation time and temperature of the competent auxotroph and DNA preparation, overnight incubation temperature, and variations in the age of the auxotroph culture when used. Under these conditions, the transformation can easily be performed in 24 h, the final 16 to 18 h being an overnight uninterrupted incubation period. When used in conjunction with the glucose oxidative fermentative basal metabolism test, it provided a rapid highly efficient means for grouping and identifying acinetobacters which is far superior to a biochemical schema. Without exception, the 141 strains of DNA from Acinetobacter species were able to transform the auxotroph to prototrophy. None of the 105 oxidase-positive nonfermenters possessed DNA which was able to transform the Acinetobacter auxotroph to prototrophy.
采用朱尼建立的方法,对250株需氧、非发酵、革兰氏阴性球杆菌和杆菌的脱氧核糖核酸(DNA)进行了测试,以检测其将不动杆菌稳定的感受态营养缺陷型菌株(trp E 27菌株)转化为原养型的能力。对朱尼的原始程序进行了若干修改,以使其适用于临床诊断实验室。这些修改主要是为了缩短程序,以便能在与当前程序一致的时间框架内完成。修改内容包括灭菌温度、感受态营养缺陷型菌株和DNA制剂的孵育时间及温度、过夜孵育温度的变化,以及使用时营养缺陷型菌株培养物菌龄的变化。在这些条件下,转化可在24小时内轻松完成,最后16至18小时为过夜不间断孵育期。当与葡萄糖氧化发酵基础代谢试验结合使用时,它为不动杆菌的分组和鉴定提供了一种快速高效的方法,远优于生化模式。无一例外,来自不动杆菌属的141株DNA菌株均能将营养缺陷型菌株转化为原养型。105株氧化酶阳性非发酵菌中,没有一株的DNA能够将不动杆菌营养缺陷型菌株转化为原养型。