Mehal W Z, Esiri M M, Lo Y M, Chapman R W, Fleming K A
Nuffield Department of Bacteriology and Pathology, John Radcliffe Hospital, Oxford.
J Clin Pathol. 1993 Jul;46(7):646-9. doi: 10.1136/jcp.46.7.646.
To develop a sensitive and specific polymerase chain reaction (PCR) based system for detecting genomic variation in JC virus. To apply this system to formalin fixed, paraffin wax embedded brain tissue from patients with and without progressive multifocal leucoencephalopathy (PML).
A pair of primers (JC1 and JC2) were designed to be complementary to the early and late regions of JC and BK polyomaviruses, respectively. A third primer (JC3), internal to JC1 and JC2, was designed to be specific for JC virus. The specificity of JC3 was investigated by amplifying plasmids with BK or JC virus genomes. Sensitivity was estimated by titration of a plasmid containing JC virus genome. Seven brains from patients with PML (PMLB) and 30 from patients without PML (non-PMLB) were amplified using JC1 and JC2, followed by JC1 and JC3. Amplification of the beta globin gene was used as an amplification control.
Amplification with JC1 and JC2 was common for JC and BK viruses, but with JC1 and JC3 it was specific for JC virus. The sensitivity of the system was 25 copies of JC plasmid per 10 microliters of digested tissue. Five out of seven PMLB and 28 of the 30 non-PMLB amplified for beta globin, but only the PMLB gave a signal with polyoma primers. Hypervariation of the length of the regulatory region of the JC isolates in the PML tissues was consistent with the presence of multiple strains of JC.
Variation in the regulatory region of JC virus can be specifically and sensitively detected from routinely processed, paraffin wax embedded brain tissue. Variation in the regulatory region is common in PML derived JC strains, but JC virus was not detectable in non-PMLB tissue.
开发一种基于聚合酶链反应（PCR）的灵敏且特异的系统，用于检测JC病毒的基因组变异。将该系统应用于患有和未患有进行性多灶性白质脑病（PML）患者的福尔马林固定、石蜡包埋脑组织。
设计一对引物（JC1和JC2），分别与JC和BK多瘤病毒的早期和晚期区域互补。设计第三个引物（JC3），位于JC1和JC2内部，对JC病毒具有特异性。通过扩增含有BK或JC病毒基因组的质粒来研究JC3的特异性。通过滴定含有JC病毒基因组的质粒来估计灵敏度。使用JC1和JC2对7例PML患者的脑（PMLB）和30例非PML患者的脑（非PMLB）进行扩增，随后使用JC1和JC3。β珠蛋白基因的扩增用作扩增对照。
用JC1和JC2扩增对JC和BK病毒很常见，但用JC1和JC3扩增对JC病毒具有特异性。该系统的灵敏度为每10微升消化组织中有25个JC质粒拷贝。7例PMLB中有5例和30例非PMLB中有28例扩增出β珠蛋白，但只有PMLB在用多瘤引物时产生信号。PML组织中JC分离株调控区长度的高度变异与多种JC毒株的存在一致。
可以从常规处理的石蜡包埋脑组织中特异性且灵敏地检测JC病毒调控区的变异。调控区的变异在源自PML的JC毒株中很常见，但在非PMLB组织中未检测到JC病毒。
