细胞培养支原体感染的DNA杂交检测。

In Vitro Pub Date : 1984-05-01 DOI:10.1007/BF02619586
S Razin, M Gross, M Wormser, Y Pollack, G Glaser
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引用次数: 8

摘要

用由克隆的山羊支原体核糖体RNA基因组成的探针对可疑细胞培养物的Eco - ri酶切DNA进行Southern blot杂交,可以检测支原体对细胞培养物的感染,并鉴定支原体。探针不能与真核生物的DNA杂交。与支原体DNA的杂交模式是物种特异性的,能够识别四种最常见的支原体污染物,口腔支原体,细耳支原体,精氨酸支原体和莱氏支原体。该测试也非常敏感,可以检测到少至1ng的支原体DNA,大致相当于10(5)支原体的DNA含量。
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Detection of mycoplasmas infecting cell cultures by DNA hybridization.

Infection of cell cultures by mycoplasmas can be detected and the mycoplasma identified by Southern blot hybridization of the Eco RI-digested DNA of the suspected cell cultures with a nick-translated probe consisting of cloned ribosomal RNA genes of Mycoplasma capricolum. The probe does not hybridize with eukaryotic DNA. The hybridization pattern with mycoplasmal DNA is species specific, enabling the identification of the four most prevalent mycoplasma contaminants, Mycoplasma orale, Mycoplasma hyorhinis, Mycoplasma arginini, and Acholeplasma laidlawii. The test is also very sensitive and can detect as little as 1 ng of mycoplasmal DNA, roughly equivalent to the DNA content of 10(5) mycoplasmas.

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