Oberhofer T R, Back A E
J Clin Microbiol. 1982 Apr;15(4):625-9. doi: 10.1128/jcm.15.4.625-629.1982.
A useful method for isolating and recognizing Haemophilus ducreyi from chancres and buboes of male patients is presented. A total of 41 clinical isolates of H. ducreyi were recovered from 33 patients over an 8-year period, and the experience with the 15 most recent isolates is presented in detail. Chocolate agar supplemented with 1% Iso VitaleX and 5% sheep blood agar were prepared, using Trypticase soy and Mueller-Hinton Agar bases, and incubation conditions included ambient, capneic, and anaerobic environments. Mueller-Hinton agar was clearly superior over Trypticase soy agar for isolation of H. ducreyi, although there was little difference between 5% sheep blood and supplemented chocolate agar. Growth in ambient air and under anaerobiasis was poor or lacking, whereas growth in 5 to 7% CO2 was good to luxuriant. Heat-inactivated and fresh (unheated)human blood clot tubes also were used for selective isolation. Although the rates of isolation from the two types of clot tube were not significantly different, unheated clot tubes were superior to heated clot tubes because of reduced level of contaminants. Gram stain characteristics taken from blood clot tubes and solid media, cellular and colonial morphology of the bacilli, and lack of oxidase, catalase, and biochemical activity except nitrate reductase were determinant factors. The results of this study demonstrated that successful isolation of H. ducreyi can be achieved with a minimal amount of resources and expertise.
本文介绍了一种从男性患者的溃疡和腹股沟淋巴结炎中分离和识别杜克雷嗜血杆菌的有效方法。在8年期间，从33例患者中总共分离出41株杜克雷嗜血杆菌临床菌株，并详细介绍了对最近15株菌株的研究经验。使用胰蛋白胨大豆琼脂和穆勒-欣顿琼脂基础培养基制备添加1%异维生素K和5%羊血的巧克力琼脂，培养条件包括常温、含二氧化碳环境和厌氧环境。虽然5%羊血琼脂和添加异维生素K的巧克力琼脂之间差异不大，但穆勒-欣顿琼脂在分离杜克雷嗜血杆菌方面明显优于胰蛋白胨大豆琼脂。在常温空气中和厌氧条件下生长较差或不生长，而在5%至7%二氧化碳环境中生长良好至茂盛。热灭活和新鲜（未加热）人血凝块管也用于选择性分离。虽然从两种类型的血凝块管中分离的比率没有显著差异，但未加热的血凝块管由于污染物水平较低而优于加热的血凝块管。从血凝块管和固体培养基中获取的革兰氏染色特征、杆菌的细胞和菌落形态，以及除硝酸盐还原酶外缺乏氧化酶、过氧化氢酶和生化活性是决定性因素。本研究结果表明，只需少量资源和专业知识就能成功分离杜克雷嗜血杆菌。
