Co-detection and discrimination of JCV and BKV DNA by duplex nested-PCR in renal transplant recipients.
OBJECTIVE
Several studies have disclosed a correlation between human polyomavirus BK (BKV) and interstitial nephritis in renal transplant recipients. It has recently been hypothesized that some cases of nephropathy may be associated with human polyomavirus JC (JCV).


METHODS
In this paper we describe the development of duplex nested-PCR assay which allows the simultaneous detection and discrimination of genomic sequences of JCV and BKV ''large T antigen'', resulting in amplicons of 150 bp and 278 bp, respectively. Thus, the presence of JCV and BKV DNA in urine and serum samples from 51 renal transplant recipients and 29 healthy controls was investigated and related to immunosuppressive regimens and renal function.


RESULTS
The comparison between the incidence of the of BKV and/or JCV infections (detected by viruria and/or viraemia) in renal transplant recipients and the control group revealed a highly significant increase of the incidence of BKV infection in immunosuppressed patients vs healthy subjects (62.7% vs 27.6%; p=0.005). In particular, we found a significant increase of BKV-DNA viruria in renal transplant recipients vs healthy subjects (49% vs 17.2%; p=0.01), in agreement with the BKV urinary shedding in renal transplant recipients of the literature (5-45%).


CONCLUSIONS
The nested-PCR technique is a valid diagnostic tool to detect viral presence in urine and its systemic diffusion. Our assay links the high sensitivity of nested amplification with the simultaneous detection and discrimination of genomic sequences of JC and BK polyomaviruses and thus provides a handy, rapid and sensitive means for DNA analysis of large numbers of samples.
Merlino
C
C
Virology Unit, Department of Public Health and Microbiology, University of Turin,Turin, Italy. chiara.merlino@unito.it
Bergallo
M
M
Daniele
R
R
Scutera
S
S
Giacchino
F
F
Comune
L
L
Negro Ponzi
A
A
Cavallo
R
R
D000328:Adult; D001739:BK Virus / Q000235:genetics*; D016022:Case-Control Studies; D004279:DNA, Viral / Q000097:blood* / Q000652:urine*; D005260:Female; D006801:Humans; D015994:Incidence; D007577:JC Virus / Q000235:genetics*; D016030:Kidney Transplantation; D008297:Male; D008875:Middle Aged; D016133:Polymerase Chain Reaction; D027601:Polyomavirus Infections / Q000453:epidemiology; D012680:Sensitivity and Specificity; D014412:Tumor Virus Infections / Q000453:epidemiology
D004279:DNA, Viral