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首页> 外文期刊>The Journal of molecular diagnostics: JMD >Diagnostic testing for pandemic influenza in Singapore: a novel dual-gene quantitative real-time RT-PCR for the detection of influenza A/H1N1/2009.
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Diagnostic testing for pandemic influenza in Singapore: a novel dual-gene quantitative real-time RT-PCR for the detection of influenza A/H1N1/2009.

机译：新加坡大流行性流感的诊断检测：一种新型双基因实时定量RT-PCR检测A / H1N1 / 2009型流感。

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摘要

With the relative global lack of immunity to the pandemic influenza A/H1N1/2009 virus that emerged in April 2009 as well as the sustained susceptibility to infection, rapid and accurate diagnostic assays are essential to detect this novel influenza A variant. Among the molecular diagnostic methods that have been developed to date, most are in tandem monoplex assays targeting either different regions of a single viral gene segment or different viral gene segments. We describe a dual-gene (duplex) quantitative real-time RT-PCR method selectively targeting pandemic influenza A/H1N1/2009. The assay design includes a primer-probe set specific to only the hemagglutinin (HA) gene of this novel influenza A variant and a second set capable of detecting the nucleoprotein (NP) gene of all swine-origin influenza A virus. In silico analysis of the specific HA oligonucleotide sequence used in the assay showed that it targeted only the swine-origin pandemic strain; there was also no cross-reactivity against a wide spectrum of noninfluenza respiratory viruses. The assay has a diagnostic sensitivity and specificity of 97.7% and 100%, respectively, a lower detection limit of 50 viral gene copies/PCR, and can be adapted to either a qualitative or quantitative mode. It was first applied to 3512 patients with influenza-like illnesses at a tertiary hospital in Singapore, during the containment phase of the pandemic (May to July 2009).
机译：2009年4月出现的全球相对缺乏对大流行性A / H1N1 / 2009病毒的免疫力以及对感染的持续敏感性，因此快速准确的诊断分析对于检测这种新型A型流感变体至关重要。在迄今为止已开发的分子诊断方法中，大多数是在串联单重测定中，其靶向单个病毒基因区段的不同区域或不同病毒基因区段。我们描述了选择性针对大流行性流感A / H1N1 / 2009的双基因（双链）定量实时RT-PCR方法。分析设计包括仅对这种新型A型流感病毒的血凝素（HA）基因具有特异性的引物探针组，以及能够检测所有猪源性A型流感病毒的核蛋白（NP）基因的第二组引物。在计算机分析中，分析中使用的特定HA寡核苷酸序列显示，它仅针对猪源性大流行毒株;对多种非流感呼吸道病毒也没有交叉反应。该测定法的诊断灵敏度和特异性分别为97.7％和100％，下限为50个病毒基因拷贝/ PCR，可以适应定性或定量模式。在大流行的遏制阶段（2009年5月至2009年7月），该方法首次应用于新加坡一家三级医院的3512例流感样疾病患者。

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