The use of unprocessed urine samples for detecting and monitoring BK viruses in renal transplant recipients by a quantitative real-time PCR assay
The use of unprocessed urine samples for detecting and monitoring BK viruses in renal transplant recipients by a quantitative real-time PCR assay
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DOI：
10.1016/j.jviromet.2007.12.015
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发表时间：
2008-04-01
影响因子：
3.1
通讯作者：
Preiksaitis, Jutta K.
中科院分区：
文献类型：
--
作者：
Pang, Xiaoli L.;Martin, Kimberly;Preiksaitis, Jutta K.
使用两种类型的样本（提取的尿液DNA和未处理的尿液），比较使用实时定量PCR（rt-QPCR）定量BK病毒载量的结果。在未处理尿液和提取的尿液DNA样本之间的定量病毒载量中观察到良好的相关性。(R-2= 0.96，p < 0.001）。与提取的尿液DNA相比，当使用小样本体积的未处理的尿液时（每个PCR反应2 μ l），在从肾移植受者（RTR）收集的124个样本（106个阳性和18个阴性）中观察到100%的一致性。与未处理尿液（中位数= 7.17）相比，在提取的尿液DNA（中位数= 7.82）中检测到的定量BK病毒载量（log（10）拷贝/ml）无显著差异。尿液pH在5.2-7.1范围内和标本冷冻对rt-QPCR反应没有影响。当使用5 μ l样品体积的未处理尿液时观察到的rt-QPCR反应的部分抑制在2 μ l样品体积时显著降低。使用未经处理的尿液进行BK病毒载量的rt-QPCR检测节省了成本，同时保持了与使用提取的尿液DNA相关的灵敏度和准确性，使得基于使用rt-QPCR作为一线检测的尿液样本筛查的RTR中的临床BKV监测策略更加可行。(C)2008 Elsevier B. V.保留所有权利。
Results of quantitative BK viral load using real-time quantitative PCR (rt-QPCR) were compared using two types of samples, extracted urine DNA and unprocessed urine. An excellent correlation was observed in quantitative viral load between unprocessed urine and extracted urine DNA samples. (R-2 = 0.96, p < 0.001). Compared to extracted urine DNA when a small sample volume of unprocessed urine was used (2 mu l per PCR reaction), 100% concordance is detection of BKV DNA was observed in 124 samples (106 positive and 18 negative) collected from renal transplant recipient (RTR). There was no significant difference in the quantitative BK viral load (log(10) copies/ml) detected in extracted urine DNA (median = 7.82) compared to unprocessed urine (median = 7.17). Urine pH in the range of 5.2-7.1 and specimen freezing had no effect on the rt-QPCR reaction. The partial inhibition of the rt-QPCR reaction observed when 5 mu l sample volume of unprocessed urine was used was markedly reduced at a sample volume of 2 mu l. Using unprocessed urine for rt-QPCR detection of BK viral load is cost-saving while maintaining the sensitivity and accuracy associated with the use of extracted urine DNA, making a clinical BKV surveillance strategy in RTR based on urinary sample screening using rt-QPCR as the first line test more feasible. (C) 2008 Elsevier B.V. All rights reserved.