Department of Respiratory Diseases Research, Naval Health Research Center, San Diego, California, United States of America.
PLoS One. 2010 Feb 3;5(2):e8995. doi: 10.1371/journal.pone.0008995.
For more than four decades the cause of most type A influenza virus infections of humans has been attributed to only two viral subtypes, A/H1N1 or A/H3N2. In contrast, avian and other vertebrate species are a reservoir of type A influenza virus genome diversity, hosting strains representing at least 120 of 144 combinations of 16 viral hemagglutinin and 9 viral neuraminidase subtypes. Viral genome segment reassortments and mutations emerging within this reservoir may spawn new influenza virus strains as imminent epidemic or pandemic threats to human health and poultry production. Traditional methods to detect and differentiate influenza virus subtypes are either time-consuming and labor-intensive (culture-based) or remarkably insensitive (antibody-based). Molecular diagnostic assays based upon reverse transcriptase-polymerase chain reaction (RT-PCR) have short assay cycle time, and high analytical sensitivity and specificity. However, none of these diagnostic tests determine viral gene nucleotide sequences to distinguish strains and variants of a detected pathogen from one specimen to the next. Decision-quality, strain- and variant-specific pathogen gene sequence information may be critical for public health, infection control, surveillance, epidemiology, or medical/veterinary treatment planning. The Resequencing Pathogen Microarray (RPM-Flu) is a robust, highly multiplexed and target gene sequencing-based alternative to both traditional culture- or biomarker-based diagnostic tests. RPM-Flu is a single, simultaneous differential diagnostic assay for all subtype combinations of type A influenza viruses and for 30 other viral and bacterial pathogens that may cause influenza-like illness. These other pathogen targets of RPM-Flu may co-infect and compound the morbidity and/or mortality of patients with influenza. The informative specificity of a single RPM-Flu test represents specimen-specific viral gene sequences as determinants of virus type, A/HN subtype, virulence, host-range, and resistance to antiviral agents.
四十多年来，大多数人类 A 型流感病毒感染的病因仅归因于两种病毒亚型，即 A/H1N1 或 A/H3N2。相比之下，禽类和其他脊椎动物是 A 型流感病毒基因组多样性的宿主，携带至少 144 种 16 种血凝素和 9 种神经氨酸酶亚型组合中的 120 种病毒株。在这个宿主中出现的病毒基因组片段重配和突变可能会产生新的流感病毒株，对人类健康和家禽生产构成迫在眉睫的流行或大流行威胁。传统的检测和区分流感病毒亚型的方法要么耗时耗力（基于培养），要么灵敏度极低（基于抗体）。基于逆转录-聚合酶链反应（RT-PCR）的分子诊断检测具有较短的检测周期时间，以及高的分析灵敏度和特异性。然而，这些诊断检测都没有确定病毒基因核苷酸序列，无法区分从一个样本到下一个样本的检测病原体的菌株和变体。区分检测病原体的菌株和变体的决策质量、菌株特异性和病原体基因序列信息对于公共卫生、感染控制、监测、流行病学或医疗/兽医治疗计划可能至关重要。重测序病原体微阵列（RPM-Flu）是一种强大的、高度多重化的基于目标基因测序的替代传统的培养或生物标志物诊断检测方法。RPM-Flu 是一种单一的、同时的差异化诊断检测方法，用于检测所有 A 型流感病毒亚型组合，以及其他 30 种可能引起流感样疾病的病毒和细菌病原体。RPM-Flu 的这些其他病原体靶点可能会合并感染并加重流感患者的发病率和/或死亡率。单一 RPM-Flu 检测的信息特异性代表了病毒类型、A/HN 亚型、毒力、宿主范围和对抗病毒药物的耐药性等决定病毒的特定样本基因序列。
