HIV-1 and HCV detection in dried blood spots by SYBR Green multiplex real-time RT-PCR
HIV-1 and HCV detection in dried blood spots by SYBR Green multiplex real-time RT-PCR
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DOI：
10.1016/j.jviromet.2009.12.017
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发表时间：
2010-04-01
影响因子：
3.1
通讯作者：
Gibellini, Davide
中科院分区：
文献类型：
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作者：
De Crignis, Elisa;Re, Maria Carla;Gibellini, Davide
干血斑（DBS）是一种可靠的血液采集方法，用于诊断几种人类疾病。脑起搏器在诊断儿童和筛查高危人群方面特别有用，特别是在卫生设施不容易获得的国家。本报告描述了一种定性的基于SYBR green的实时多重RT-PCR，用于同时检测DBS中丙型肝炎病毒（HCV）和人类免疫缺陷病毒1型（HIV-1）基因组。在同一样品中，通过其独特的熔化温度鉴定出特定的病毒扩增子。参考样品的标量浓度分析表明，这种多重程序检测到至少2500拷贝/ml的HCV和400拷贝/ml的HIV-1。对20名感染HIV-1和/或HCV的患者和5名健康献血者的HIV-1和HCV病毒载量进行了检测，证实了该检测方法的敏感性和特异性。在快速和相对廉价的筛查方案中，这种方法可能是检测HIV-1/HCV合并感染的可靠替代方法。(C) 2010 Elsevier B.V.版权所有
Dried blood spot (DBS) is a reliable method of blood collection used for the diagnosis of several human diseases. DBS is particularly useful for diagnosing children and for the screening of high-risk populations especially in countries where health facilities are not readily accessible. This report describes a qualitative SYBR Green-based real-time multiplex RT-PCR for the simultaneous detection of hepatitis C virus (HCV) and human immunodeficiency virus type 1 (HIV-1) genomes in DBS. Specific viral amplicons were identified in the same sample by their distinctive melting temperatures. The analysis of scalar concentrations of the reference samples indicated that this multiplex procedure detects at least 2500 copies/ml of HCV and 400 copies/ml of HIV-1. HIV-1 and HCV viral loads in 20 patients infected with HIV-1 and/or HCV and in 5 healthy blood donors were also tested, confirming the sensitivity and specificity of the assay.This method may represent a reliable alternative for the detection of HIV-1/HCV co-infection, in rapid and relatively inexpensive screening programmes. (C) 2010 Elsevier B.V. All rights reserved.