应用 SYBR Green 多重实时 RT-PCR 法检测干血斑中的 HIV-1 和 HCV。
HIV-1 and HCV detection in dried blood spots by SYBR Green multiplex real-time RT-PCR.
机构信息
Department of Haematology and Oncologic Sciences, Section of Microbiology, Rome, Italy.
出版信息
J Virol Methods. 2010 Apr;165(1):51-6. doi: 10.1016/j.jviromet.2009.12.017. Epub 2010 Jan 5.
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.
干血斑（DBS）是一种可靠的血液采集方法，可用于诊断多种人类疾病。DBS 特别适用于诊断儿童疾病，以及对高危人群进行筛查，尤其是在那些卫生设施不易获得的国家。本报告描述了一种基于 SYBR Green 的实时多重 RT-PCR 方法，可用于同时检测干血斑中的丙型肝炎病毒（HCV）和人类免疫缺陷病毒 1 型（HIV-1）基因组。通过其独特的熔解温度，在同一样本中鉴定出特定的病毒扩增子。对参考样本的标度浓度分析表明，这种多重程序可检测到至少 2500 拷贝/ml 的 HCV 和 400 拷贝/ml 的 HIV-1。还对 20 名感染 HIV-1 和/或 HCV 的患者以及 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。该方法可能是 HIV-1/HCV 合并感染快速且相对廉价的筛查方案中可靠的替代方法。
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