结核分枝杆菌快速培养方法的建立及临床应用初步观察

Establishment on the fast cultivation method for Mycobacterium tuberculosis and its primary observation in clinical practice

目的 观察结核分枝杆菌在自制培养基上的生长情况，建立快速培养方法。方法将改良罗氏培养基的成分进行改进，自制5种培养基，每种3支，每支接种菌量10-3mg，观察结核分枝杆菌标准株生长情况。并以第5号培养基3种成分不同浓度做正交叉实验，得出最佳浓度。收集临床标本，在自制培养基上培养观察。结果 结核分枝杆菌标准株在5种自制培养基上，菌落开始生长天数及典型菌落出现天数均比改良罗氏培养基短，两者差异均具有显著性（P〈O．01）。第5号培养基以25％牛肉浸液，20％当归浸液，20％党参浸液的浓度最佳，结核分枝杆菌标准株接种后第3～5d开始生长，第7～9d出现典型的“菜花状”菌落。临床标本接种在改良罗氏及自制培养基上，阳性分离率差异无显著性（f—O．09，P〉O．05），两种培养基上典型菌落形成的平均天数差异有显著性（f一6．828，P〈O．001）。结论 结核分枝杆菌标准株在自制第5号培养基上生长快，培养时间短，为该菌培养提供了一种新的快速培养基，在临床上有一定的实用价值，值得进一步研究、推广。

Objective To establish a fast cultivation method of Mycobacterium tuberculosis （MT） through improving culture medium. Methods Culture mediums of different ingredient were prepared through improving the Lowenstein- Jensen medium and were divided into 5 groups, 3 tubes in each. Inoculation amount of MT was 10-3 mg in each tube, and the growth of MT was observed. We did a orthogonal test to obtain the best concentration by using different concentration of three ingredients of the 5th medium. Then collection of clinical specimen and observation of the growth situation of MT on the self-prepared culture medium was conducted to obtain the best cultivation method. Results The starting growth time of bacterium cluster and the appearance time of typical bacterium cluster were significantly shorter than that of Lo- wenstein-Jensen medium （P〈0. 01） when the standard strains of MT were cultured on the 5 groups of self-prepared cul- ture medium. In the 5th medium, the best concentration of different ingredient was 25% beef broth, 20% angelica and 20%Dangshen extract. In the above medium, the MT started growing after 3-5 days of incubation and the typical ＂cau- liflower-like＂ bacterium cluster appeared after 7-9 days of incubation. After clinical specimen in Lowenstein-Jensen me- dium and the 5th medium were incubated, separating rate of MT had no significance（t=0.09, P〉0.05）, but there was significant difference in average days of appearance of typical bacterium cluster （t= 6. 828, P〈0. 001 ）. Conclusion MT grew quickly on the self-prepared of the 5th medium, the incubation time was shorter. It is a new and fast culture medium for this bacterium incubation. This method has a certain practical clinical value and is worth studying further.