Dörries K, ter Meulen V
J Med Virol. 1983;11(4):307-17. doi: 10.1002/jmv.1890110406.
Cellular DNA of the kidney from a patient with PML was analyzed by reassociation kinetics for the presence of JC virus DNA. Various amounts of viral DNA sequences were detected in different areas of the kidney. The highest concentration (175 genome equivalents/cell) was found in the renal medulla and there were almost none in the renal cortex. Differentiation from the closely related BK virus was carried out by reassociation kinetics and restriction enzyme cleavage with subsequent Southern blot analysis. The enzyme Hind II, which does not cleave within the BK virus genome, generated four restriction enzyme fragments in the cellular DNA from the kidney, thus documenting the presence of JC virus DNA. By examination of the renal DNA with the "no-cut" restriction enzyme XHO I and the "one-cut" enzymes Eco RI and BAM HI it was possible to show that free and not integrated viral DNA was present in these cells. Nonhomogeneous defective DNA bands were not detectable. By in situ hybridization the epithelial cells lining the collecting tubules were found as predominant site of the viral infection in the kidney.
对一名进行性多灶性白质脑病（PML）患者肾脏的细胞DNA进行重缔合动力学分析，以检测是否存在JC病毒DNA。在肾脏的不同区域检测到了不同数量的病毒DNA序列。在肾髓质中发现最高浓度（175个基因组当量/细胞），而肾皮质中几乎没有。通过重缔合动力学以及随后用限制性内切酶切割并进行Southern印迹分析，与密切相关的BK病毒进行区分。Hind II酶在BK病毒基因组内不切割，在肾脏细胞DNA中产生了四个限制性酶切片段，从而证明了JC病毒DNA的存在。通过用“不切割”的限制性内切酶XHO I以及“单切割”的酶Eco RI和BAM HI检测肾脏DNA，有可能表明这些细胞中存在游离而非整合的病毒DNA。未检测到非均一性缺陷DNA条带。通过原位杂交发现，集合小管内衬的上皮细胞是肾脏中病毒感染的主要部位。
