A simple biochemical method for the detection of mycoplasmas and other microbial contaminants of cell cultures
A simple biochemical method for the detection of mycoplasmas and other microbial contaminants of cell cultures
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检测细胞培养物支原体和其他微生物污染物的简单生化方法
DOI：
10.1007/bf00921623
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发表时间：
1976
期刊：
影响因子：
--
通讯作者：
E. Schneider
中科院分区：
文献类型：
--
作者：
E. Stanbridge;E. Schneider
支原体污染物对细胞培养人员的危害已得到充分证明（1）。由于在解释支原体污染细胞的数据时存在问题，大多数细胞生物学家常规筛选培养物中是否存在这些微生物。目前，最广泛使用的支原体检测技术包括在专门制备的肉汤和固体琼脂中培养这些微生物。支原体的阳性鉴定需要出现特征性的“煎蛋”菌落。支原体标准微生物检测的局限性越来越明显;现在人们认识到，有些难养支原体即使不是不可能，也极难在目前可用的无细胞培养基上生长（2）。因此，已经开发了几种非培养试验来帮助检测这些污染物。在本节中，我们描述了一种简单的生化方法，几乎可以在任何实验室进行。这是尿苷：尿嘧啶比率方法，其基于在支原体感染细胞中观察到的两种生化改变：外源性尿嘧啶（U）掺入增加;和外源性尿苷（Urd）掺入核糖核酸（RNA）中减少（3）。通过利用放射性标记的化合物测量Urd和U掺入RNA。
The hazards that mycoplasma contaminants present to cell culturists are well documented (1). Because of the problems in interpreting data derived from mycoplasma contaminated cells, most cell biologists routinely screen their cultures for the presence of these organisms. Currently, the most widely used technique for mycoplasma detection involves growth of these organisms in specially prepared broth and on solid agar. Positive identification of mycoplasmas requires the appearance of characteristic "fried egg" colonies. The limitations of standard microbiological testing for mycoplasmas have become increasingly apparent; and it is now recognized that there are fastidious mycoplasmas that are extremely difficult, if not impossible, to grow on currently available acellular media (2). Consequently several noncultural tests have been developed to assist in the detection of these contaminants. In this section we describe a simple biochemical method which can be performed in almost any laboratory. This is the uridine:uracil ratio method, which is based on two biochemical alterations that are observed in mycoplasma infected cells: increased incorporation of exogenous uracil (U); and decreased incorporation of exogenous uridine (Urd) into ribonucleic acid (RNA) (3). The incorporation of Urd and U into RNA is measured by utilizing radioactively labeled compounds.