Huang Ying, Tang Huong, Duffy Stuart, Hong Yuwen, Norman Sylvia, Ghosh Madhu, He Jie, Bose Michael, Henrickson Kelly J, Fan Jiang, Kraft Andrea J, Weisburg William G, Mather Elizabeth L
Nanogen Inc, San Diego, CA 92121.
J Clin Microbiol. 2009 Feb;47(2):390-6. doi: 10.1128/JCM.01807-08. Epub 2008 Dec 10.
We report on the use of an electronic microarray to simultaneously type influenza A and B viruses and to distinguish influenza A virus subtypes H1N1 and H3N2 from the potentially pandemic avian virus subtype H5N1. The assay targets seven genes: the H1, H3, H5, N1, and N2 genes of influenza A virus; the matrix protein M1 gene of influenza A virus; and the nonstructural protein (NS) gene of influenza B virus. By combining a two-step reverse transcription-multiplex PCR with typing and subtyping on the electronic microarray, the assay achieved an analytical sensitivity of 10(2) to 10(3) copies of transcripts per reaction for each of the genes. The assay correctly typed and subtyped 15 different influenza virus isolates, including two influenza B virus, five A/H1N1, six A/H3N2, and two A/H5N1 isolates. In addition, the assay correctly identified 8 out of 10 diluted, archived avian influenza virus specimens with complete typing and subtyping information and 2 specimens with partial subtyping information. In a study of 146 human clinical specimens that had previously been shown to be positive for influenza virus or another respiratory virus, the assay showed a clinical sensitivity of 96% and a clinical specificity of 100%. The assay is a rapid, accurate, user-friendly method for simultaneously typing and subtyping influenza viruses.
我们报告了一种电子微阵列的应用,该微阵列可同时对甲型和乙型流感病毒进行分型,并将甲型流感病毒H1N1和H3N2亚型与具有潜在大流行风险的禽流感病毒H5N1亚型区分开来。该检测方法针对七个基因:甲型流感病毒的H1、H3、H5、N1和N2基因;甲型流感病毒的基质蛋白M1基因;以及乙型流感病毒的非结构蛋白(NS)基因。通过将两步逆转录-多重PCR与电子微阵列上的分型和亚型分析相结合,该检测方法对每个基因的分析灵敏度达到了每个反应10(2)至10(3)个转录本拷贝。该检测方法对15种不同的流感病毒分离株进行了正确的分型和亚型分析,包括两种乙型流感病毒、五种A/H1N1、六种A/H3N2和两种A/H5N1分离株。此外,该检测方法正确鉴定了10份具有完整分型和亚型信息的稀释存档禽流感病毒标本中的8份,以及2份具有部分亚型信息的标本。在一项对146份先前已被证明对流感病毒或其他呼吸道病毒呈阳性的人类临床标本的研究中,该检测方法的临床灵敏度为96%,临床特异性为100%。该检测方法是一种快速、准确、用户友好的同时对流感病毒进行分型和亚型分析的方法。