Real-time RT-PCR assays for discriminating influenza B virus Yamagata and Victoria lineages.
Real-time RT-PCR assays for discriminating influenza B virus Yamagata and Victoria lineages.
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DOI：
10.1016/j.jviromet.2014.04.016
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发表时间：
2014-09-01
影响因子：
3.1
通讯作者：
Kageyama T
中科院分区：
文献类型：
--
作者：
Nakauchi M;Takayama I;Takahashi H;Oba K;Kubo H;Kaida A;Tashiro M;Kageyama T
我们开发了一步实时RT-PCR检测方法来区分两个乙型流感病毒谱系。使用体外转录对照RNA、临床标本和临床分离株对所开发的检测方法进行了评估。结果表明，该方法具有高灵敏度和高特异性。检测结果与血凝抑制试验（HI）的结果一致，后者是确定乙型流感病毒谱系的标准方法。所开发的检测方法将有助于B型流感病毒的诊断和监测。自20世纪80年代末以来，两种基因和抗原性不同的乙型流感病毒谱系，即B/Victoria/2/87样（B/Victoria）和B/Yamagata/16/88样（B/Yamagata）共同传播。在这项研究中，建立了一步实时逆转录pcr （rRT-PCR）方法来区分B/Victoria和B/Yamagata谱系。使用体外转录的对照RNA、分离的病毒和其他呼吸道病原病毒对检测结果进行了评价，结果表明检测结果灵敏度高、线性良好（R2 = 0.99）、特异性高。利用开发的rRT-PCR检测方法，对2010年至2013年期间收集的169份临床标本进行了检测，结果确定20份临床标本呈乙型流感病毒阳性。其中，14个和6个样本分别被确定为B/Victoria和B/Yamagata谱系阳性，而149个样本对乙型流感病毒呈阴性。对20例乙型流感病毒阳性标本中的20株临床分离株进行了rRT-PCR检测，结果显示rRT-PCR检测结果与血凝抑制（HI）试验结果无差异，但HI试验无法区分具有不同抗原性的一株临床分离株。目前的结果表明，这些高灵敏度和特异性的检测方法不仅可用于流感病毒的诊断，而且可用于流感病毒的监测。
We developed one step real-time RT-PCR assays to discriminate two lineages of influenza B viruses. The developed assays were evaluated using in vitro transcribed control RNA, clinical specimens, and clinical isolates. The assays were shown to have high sensitivity and high specificity. The results from the assays were consistent with those from a hemagglutination inhibition (HI) test, which is a standard method to define the lineage of influenza B virus. The developed assays will be useful for the diagnosis and surveillance of influenza B viruses. Since the late 1980s, two genetically and antigenically distinct lineages of influenza B virus, namely, B/Victoria/2/87-like (B/Victoria) and B/Yamagata/16/88-like (B/Yamagata), have co-circulated. In this study, one-step real-time reverse transcription-PCR (rRT-PCR) assays were developed to differentiate B/Victoria and B/Yamagata lineages. The assays were evaluated using in vitro transcribed control RNA, isolated viruses, and other respiratory pathogenic viruses, and were shown to have high sensitivity, good linearity (R2 = 0.99), and high specificity. Using the developed rRT-PCR assays, 169 clinical specimens collected between 2010 and 2013 were then tested, resulting in the identification of 20 clinical specimens as positive for influenza B virus. Of these, 14 and 6 samples were identified as positive for the B/Victoria and B/Yamagata lineages, respectively, whereas 149 samples were negative for the influenza B virus. The rRT-PCR assays were also examined using 20 clinical isolates from 20 influenza B virus-positive specimens, revealing that there was no discrepancy between the results from the rRT-PCR assays and the hemagglutination inhibition (HI) test, with the exception that one clinical isolate with different antigenicity could not be discriminated by the HI test. The present results suggest that these highly sensitive and specific assays are useful not only for diagnosing influenza viruses but also for their surveillance.
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影响因子：
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