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用于鉴定引起人类急性呼吸道感染的病毒病原体的实时聚合酶链反应检测方法的开发与验证

[Development and validation of the real-time PCR assay for the identification of viral agents causing acute respiratory infections in humans].

作者信息

Sergeeva E I, Ternovoĭ V A, Demina O K, Demina A V, Korneev A N, Shikov A N, Berillo S A, Agafonov A P, Sergeev A N

出版信息

Mol Gen Mikrobiol Virusol. 2013(4):32-7.

Abstract

The real time PCR assay targeting influenza A and B virus, 5 subtypes of influenza A virus (seasonal H1N1, pandemic H1N1 (2009), seasonal H3N2, pathogenic for human subtypes of avian influenza H5 and H7), respiratory syncytial virus, and adenovirus was developed. The analytical sensitivity of the developed assay was 1 x 10(3) genome equivalents per ml. The diagnostic sensitivity of the method was 1 x l0(3)-10(4) viral particles per ml. Experiments with human DNA/cDNA and viral cDNA showed a markedly high diagnostic specificity of the developed PCR assay. In the assay of the developed PCR test, 50 nasopharyngeal swab specimens were tested. The etiology was identified in 33 samples.

摘要

开发了一种针对甲型和乙型流感病毒、甲型流感病毒的5种亚型(季节性H1N1、大流行H1N1(2009年)、季节性H3N2、对人类致病的禽流感H5和H7亚型)、呼吸道合胞病毒和腺病毒的实时PCR检测方法。所开发检测方法的分析灵敏度为每毫升1×10³个基因组当量。该方法的诊断灵敏度为每毫升1×10³ - 10⁴个病毒颗粒。用人DNA/cDNA和病毒cDNA进行的实验表明,所开发的PCR检测方法具有显著高的诊断特异性。在所开发的PCR检测试验中,对50份鼻咽拭子标本进行了检测。在33份样本中确定了病因。

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