Evaluation of different urine protocols and DNA extraction methods for quantitative detection of BK viruria in kidney transplant patients

被引：9
|
作者：
Pinto, Gabriel Godinho [1 ]
Tesser Poloni, Jose Antonio [1 ,2 ]
Carneiro, Lilian Carla [1 ]
Baethgen, Ludmila Fiorenzano [1 ]
Barth, Afonso Luis [3 ,4 ]
Pasqualotto, Alessandro Comaru [1 ,5 ]
机构：
[1] UFCSPA, Porto Alegre, RS, Brazil
[2] Santa Casa de Misericordia Porto Alegre, Cent Lab, Porto Alegre, RS, Brazil
[3] HCPA, Serv Patol Clin, Porto Alegre, RS, Brazil
[4] Univ Fed Rio Grande do Sul, Fac Farm, Porto Alegre, RS, Brazil
[5] Santa Casa de Misericordia Porto Alegre, Mol Biol Lab, Porto Alegre, RS, Brazil
来源：
JOURNAL OF VIROLOGICAL METHODS | 2013年 / 188卷 / 1-2期
关键词：
BKV; Diagnosis; Kidney transplantation; Polyomavirus; Real-time polymerase chain reaction; POLYMERASE-CHAIN-REACTION; TIME PCR ASSAY; REPLICATION; RECIPIENTS;
D O I：
10.1016/j.jviromet.2012.12.006
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
Most transplant centers screen kidney transplant recipients for BK virus (BKV) infection using molecular techniques for the virus load determination. However, there is no consensus about the pre-analytical methods involved in the viral detection. In this study BK viral load was compared by the means of two urine treatment protocols (pelleted vs. whole urine) and two commercial DNA extraction kits for a quantitative PCR (qPCR) experiment. Ten patients who presented decoy cells in their urine sediment were selected for the study. Viral load was considerable higher (>1.5 log) for pelleted urine, in comparison to whole urine but no significant difference was observed between the extraction kits. PCR inhibition did not occur by using pelleted urine. In order to increase test sensitivity to detect BK viruria, pelleted urine should be the preferred urine compartment for qPCR experiments. (C) 2013 Elsevier B.V. All rights reserved.
引用
收藏
页码：94 / 96
页数：3
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