使用 QIAGEN Artus BK 病毒 RG PCR 检测评估 BK 病毒载量分析。

Evaluation of a BK virus viral load assay using the QIAGEN Artus BK Virus RG PCR test.

机构信息

Department of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, NY 10065, USA.

出版信息

J Clin Virol. 2012 Jul;54(3):260-4. doi: 10.1016/j.jcv.2012.03.007. Epub 2012 Apr 10.

DOI:10.1016/j.jcv.2012.03.007

PMID:22494899

Abstract

BACKGROUND

Viral load testing for BK Virus (BKV) has become the standard of care for the diagnosis of infection and monitoring of therapy of kidney transplant patients infected with BKV. However, there are currently no FDA-approved BKV quantification assays and no standardization among available tests.

OBJECTIVE AND STUDY DESIGN

This study evaluated the performance of the Artus BK Virus RG PCR (RUO) assay (QIAGEN) for accuracy, linearity, precision, analytical sensitivity, specificity, and correlation with a referral laboratory test in patient samples.

RESULTS

Linear regression analysis of the quantitative results demonstrated a linear range of quantification from 192 to 194 million (2.28 to 8.29 log(10)) DNA copies/mL and a coefficient of determination (R(2)) of 0.994. A dilution series demonstrated a limit of detection and a limit of quantification of 2.00 log(10), and 2.30 log(10) copies/mL (>95% positivity rate), respectively. The precision of the assay was highly reproducible among runs with coefficients of variance (CV) ranging from 0.2% to 7.0%. A comparison of 34 matched samples showed a good agreement (R(2)=0.983) between the Artus BK test and the referral laboratory results, with an average positive bias (0.39 log(10) copies/mL). Genotyping analysis using large-T antigen sequences demonstrated that 90% of the positive samples were BKV type I, and that there was no significant difference in quantification between the referral laboratory and Artus BK Virus tests.

CONCLUSIONS

The Artus BK Virus RG PCR test is a reliable and sensitive assay for BKV DNA quantification as compared to the referral laboratory test.

摘要

背景

BK 病毒（BKV）的病毒载量检测已成为诊断肾移植患者 BKV 感染和监测治疗的标准。然而，目前尚无获得 FDA 批准的 BKV 定量检测方法，并且可用检测方法之间也没有标准化。

目的和研究设计

本研究评估了 Artus BK Virus RG PCR（RUO）检测（QIAGEN）在患者样本中的准确性、线性、精密度、分析灵敏度、特异性以及与参考实验室检测的相关性。

结果

定量结果的线性回归分析表明，定量范围为 1.92 亿至 1.94 亿（2.28 至 8.29 log(10)) DNA 拷贝/mL，决定系数（R(2))为 0.994。稀释系列表明检测限和定量限分别为 2.00 log(10)和 2.30 log(10)拷贝/mL（>95%阳性率）。该检测在运行过程中的精密度高度可重现，变异系数（CV）范围为 0.2%至 7.0%。34 个匹配样本的比较表明，Artus BK 检测与参考实验室结果具有良好的一致性（R(2)=0.983），平均阳性偏倚（0.39 log(10) 拷贝/mL）。使用大 T 抗原序列进行基因分型分析表明，90%的阳性样本为 BKV 型 I，并且参考实验室和 Artus BK 病毒检测之间的定量没有显著差异。