DETECTION BY PCR OF HUMAN POLYOMAVIRUSES BK AND JC IN IMMUNOCOMPROMISED INDIVIDUALS AND PARTIAL SEQUENCING OF CONTROL REGIONS

Immunocompromised individuals were tested for the presence of the huma n polyomaviruses JC (JCV) and BK (BKV) by the polymerase chain reactio n (PCR). The use of appropriate primers in a nested PCR allowed the de tection 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 pr ogressive multifocal leukencephalopathy. In autopsy materials from thi s 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 methology further, hybridization-positive urines from three recipients of bone marrow transplants and a positive urine of an acute myeloid l eukemia patient were analyzed by this PCR method. One case was positiv e 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 wer e sequenced directly from PCR-derived fragments. The JCV sequence from urine of the AIDS patient compared to sequences from a bone marrow tr ansplant recipient and to archetypical reference strains showed two nu cleotide (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 co ntrol region and were identified as BKV WW or WWT3 strains. In the agn ogene region five exchanges were detected, two of them resulting in no n-conservative amino acid exchanges. The possibility of testing clinic al specimens of different origins by this PCR method is important for elucidating often unclear clinical courses in immunocompromised patien ts. Furthermore, our results show the versatility of PCR for diagnosis and for molecular characterization of human polyomaviruses. (C) 1994 Wiley-Liss, Inc.