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[一种在含透明质酸钾凝胶中快速检测细菌透明质酸酶的简单方法]

[A simple method for the rapid detection of bacterial hyaluronidase in K hyaluronate-containing gel].

作者信息

Balke E, Weiss R

出版信息

Zentralbl Bakteriol Mikrobiol Hyg A. 1984 Aug;257(3):317-22.

PMID:6485633
Abstract

For detection of hyaluronidase activities we investigated several groups of bacteria. The bacteria were inoculated on a 1,5% agarose gel in Petri plates of 4 cm diameter or gel discs of 7 mm diameter, containing 0,1% of K-hyaluronate as well as nutritient medium, and were incubated for 2-20 h at 37 degrees C in a moist chamber. Subsequently some ml of a 10% solution of cetylpyridiniumchloride were poured on the gel to precipitate the polymere hyaluronate. If the hyaluronate was depolymerized by hyaluronidase, a translucent area was visible around the colonies. We found out, that a gel layer of 1 mm was sufficient to detect the small amounts of hyaluronidase, which were produced by bacteria within an incubation time of 2 h. These results were confirmed by incubation for 20 h and in some cases 36 h. The hyaluronidase production by different anaerobic Clostridium strains was always proved after a 20 h growth period. The bacteria were inoculated with the whole loop of a self made platin sowing wire loop. By this method quantitative differences of hyaluronidase activities between different strains of bacteria could be detected.

摘要

为检测透明质酸酶活性,我们研究了几组细菌。将细菌接种在直径4厘米的培养皿中的1.5%琼脂糖凝胶或直径7毫米的凝胶圆盘上,凝胶中含有0.1%的钾透明质酸盐以及营养培养基,并在潮湿环境中于37℃孵育2 - 20小时。随后,将几毫升10%的十六烷基氯化吡啶溶液倒在凝胶上,以使聚合的透明质酸盐沉淀。如果透明质酸盐被透明质酸酶解聚,菌落周围会出现一个半透明区域。我们发现,1毫米厚的凝胶层足以检测细菌在2小时孵育时间内产生的少量透明质酸酶。这些结果在孵育20小时以及某些情况下孵育36小时后得到了证实。不同厌氧梭菌菌株的透明质酸酶产生情况在生长20小时后总能得到证实。用自制的铂接种环整环接种细菌。通过这种方法,可以检测不同菌株之间透明质酸酶活性的定量差异。

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