Schätzl H M, Sieger E, Jäger G, Nitschko H, Bader L, Ruckdeschel G, Jäger G
Max von Pettenkofer Institute, University of Munich, Germany.
J Med Virol. 1994 Feb;42(2):138-45. doi: 10.1002/jmv.1890420208.
Immunocompromised individuals were tested for the presence of the human polyomaviruses JC (JCV) and BK (BKV) by the polymerase chain reaction (PCR). The use of appropriate primers in a nested PCR allowed the detection of both viruses simultaneously. Viruses were differentiated by restriction fragment length analysis of amplified DNA fragments. Both BKV and JCV DNA were detected in the urine of an AIDS patient with progressive multifocal leukencephalopathy. In autopsy materials from this patient, JCV- but not BKV-DNA was found in brain and kidney tissue, whereas lung tissue was negative for both virus DNAs. To evaluate the methodology further, hybridization-positive urines from three recipients of bone marrow transplants and a positive urine of an acute myeloid leukemia patient were analyzed by this PCR method. One case was positive both for BKV and JCV, two cases were positive only for BKV, and one was negative for both. Parts of the control regions of JCV and BKV were sequenced directly from PCR-derived fragments. The JCV sequence from urine of the AIDS patient compared to sequences from a bone marrow transplant recipient and to archetypical reference strains showed two nucleotide (nt) exchanges out of 250 nt. The BKV sequences from the AML and the AIDS patients showed five nt exchanges out of 265 nt in the control region and were identified as BKV WW or WWT3 strains. In the agnogene region five exchanges were detected, two of them resulting in non-conservative amino acid exchanges. The possibility of testing clinical specimens of different origins by this PCR method is important for elucidating often unclear clinical courses in immunocompromised patients.(ABSTRACT TRUNCATED AT 250 WORDS)
通过聚合酶链反应（PCR）对免疫功能低下个体检测人多瘤病毒JC（JCV）和BK（BKV）的存在情况。在巢式PCR中使用合适的引物可同时检测这两种病毒。通过对扩增的DNA片段进行限制性片段长度分析来区分病毒。在一名患有进行性多灶性白质脑病的艾滋病患者的尿液中检测到了BKV和JCV DNA。在该患者的尸检材料中，在脑和肾组织中发现了JCV DNA而非BKV DNA，而肺组织中两种病毒DNA均为阴性。为进一步评估该方法，用此PCR方法分析了来自三名骨髓移植受者的杂交阳性尿液以及一名急性髓系白血病患者的阳性尿液。一例BKV和JCV均为阳性，两例仅BKV为阳性，一例两者均为阴性。直接从PCR扩增片段对JCV和BKV的部分控制区进行测序。艾滋病患者尿液中的JCV序列与骨髓移植受者的序列以及原型参考菌株相比，在250个核苷酸（nt）中有两个核苷酸交换。急性髓系白血病患者和艾滋病患者的BKV序列在控制区的265个nt中有五个nt交换，被鉴定为BKV WW或WWT3菌株。在无义基因区检测到五个交换，其中两个导致非保守氨基酸交换。用这种PCR方法检测不同来源临床标本的可能性对于阐明免疫功能低下患者常常不明的临床病程很重要。（摘要截短至250字）
