Merlino C, Bergallo M, Daniele R, Scutera S, Giacchino F, Comune L, Negro Ponzi A, Cavallo R
Virology Unit, Department of Public Health and Microbiology, University of Turin,Turin, Italy.
Panminerva Med. 2004 Sep;46(3):153-9.
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).
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.
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%).
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.
多项研究揭示了人类多瘤病毒BK（BKV）与肾移植受者间质性肾炎之间的相关性。最近有人推测，某些肾病病例可能与人类多瘤病毒JC（JCV）有关。
在本文中，我们描述了双重巢式聚合酶链反应检测方法的开发，该方法可同时检测和区分JCV和BKV“大T抗原”的基因组序列，分别产生150bp和278bp的扩增子。因此，我们研究了51名肾移植受者和29名健康对照者尿液和血清样本中JCV和BKV DNA的存在情况，并将其与免疫抑制方案和肾功能相关联。
肾移植受者与对照组中BKV和/或JCV感染（通过病毒尿和/或病毒血症检测）发生率的比较显示，免疫抑制患者中BKV感染的发生率与健康受试者相比显著增加（62.7%对27.6%；p=0.005）。特别是，我们发现肾移植受者中BKV-DNA病毒尿与健康受试者相比显著增加（49%对17.2%；p=0.01），这与文献中肾移植受者的BKV尿排泄情况（5%-45%）一致。
巢式聚合酶链反应技术是检测尿液中病毒存在及其全身扩散的有效诊断工具。我们的检测方法将巢式扩增的高灵敏度与JC和BK多瘤病毒基因组序列的同时检测和区分相结合，从而为大量样本的DNA分析提供了一种方便、快速且灵敏的手段。
