Real-Time Detection of Influenza A, Influenza B, and Respiratory Syncytial Virus A and B in Respiratory Specimens by Use of Nanoparticle Probes
Real-Time Detection of Influenza A, Influenza B, and Respiratory Syncytial Virus A and B in Respiratory Specimens by Use of Nanoparticle Probes
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DOI：
10.1128/jcm.01118-10
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发表时间：
2010-11-01
影响因子：
9.4
通讯作者：
Ledeboer, Nathan A.
中科院分区：
文献类型：
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作者：
Jannetto, Paul J.;Buchan, Blake W.;Ledeboer, Nathan A.
流感和呼吸道合胞病毒的季节性流行是造成全世界显着发病率和死亡率的原因。罕见情况下，新型或重新出现的甲型流感病毒株会引起快速、严重的全球大流行，导致数百万人死亡。高效、准确地检测和区分呼吸道病毒的能力对于有效治疗、感染控制和流行病学监测至关重要。我们评估了两种 FDA 批准的基于核酸的测试，即半自动呼吸道病毒核酸测试 (VRNAT) 和全自动呼吸道病毒核酸测试 SP (RVNAT(SP))（Nanosphere Inc.，Northbrook，IL）从临床鼻咽拭子样本中检测甲型流感病毒、乙型流感病毒以及呼吸道合胞病毒 A 和 B (RSV A/B) 的能力。两项测试中病毒 RNA 的检测均基于核酸扩增，然后进行杂交以捕获固定在载玻片上的探针。利用金纳米粒子缀合探针的新技术来检测捕获的目标 DNA 的存在。这种基于微阵列的检测方法已被证明比传统的培养/直接荧光抗体测定 (DFA) 方法更灵敏，可用于检测临床样本中的 RSV 和流感病毒，包括新型 2009 H1N1 病毒株。具体来说，与培养/DFA 相比，我们报告 VRNAT 的敏感性为 98.0%，特异性为 96.5%。此外，VRNAT 在另外 58% 的培养阴性样本中检测到病毒。这些数据通过双向测序得到证实。完全自动化的 RVNAT(SP) 建立在与 VRNAT 相同的检测技术之上，但包含可实现完全自动化的更新处理器，对全自动 RVNAT(SP) 的评估表明，这两个测试在功能上是等效的。因此，RVNAT(SP) 是一种全自动的从样本到结果的测试，能够在 3.5 小时内直接从临床样本中可靠地检测选定的呼吸道病毒。
Seasonal epidemics of influenza and respiratory syncytial virus are responsible for significant morbidity and mortality worldwide. Infrequently, novel or reemergent strains of influenza A virus have caused rapid, severe global pandemics resulting in millions of fatalities. The ability to efficiently and accurately detect and differentiate respiratory viruses is paramount for effective treatment, infection control, and epidemiological surveillance. We evaluated the ability of two FDA-cleared nucleic acid-based tests, the semiautomated respiratory virus nucleic acid test (VRNAT) and the fully automated respiratory virus nucleic acid test SP (RVNAT(SP)) (Nanosphere Inc., Northbrook, IL) to detect influenza A virus, influenza B virus, and respiratory syncytial virus A and B (RSV A/B) from clinical nasopharyngeal swab specimens. Detection of viral RNA in both tests is based on nucleic acid amplification followed by hybridization to capture probes immobilized on a glass slide. A novel technology utilizing gold nanoparticle-conjugated probes is utilized to detect the presence of captured target DNA. This microarray-based approach to detection has proven to be more sensitive than the traditional culture/direct fluorescent-antibody assay (DFA) method for detecting RSV and influenza viruses in clinical specimens, including the novel 2009 H1N1 strain. Specifically, we report 98.0% sensitivity and 96.5% specificity for the VRNAT compared to culture/DFA. Further, the VRNAT detected virus in an additional 58% of specimens that were culture negative. These data were confirmed using bidirectional sequencing. Evaluation of the fully automated RVNAT(SP), which is built on the same detection technology as the VRNAT but contains an updated processor enabling complete automation, revealed the two tests to be functionally equivalent. Thus, the RVNAT(SP) is a fully automated sample-to-result test capable of reliable detection of select respiratory viruses directly from clinical specimens in 3.5 h.