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本研究建立了一种基于GeXP多重基因表达遗传分析系统的多重RT-PCR检测方法,该方法可以同时检测12种呼吸道病毒,包括流感病毒A型和B型、季节性H1N1、副流感病毒1～3型、人鼻病毒、人偏肺病毒、腺病毒、呼吸道合胞病毒A型和B型、人博卡病毒。针对病原体保守区序列设计12种病毒的特异性引物,分别用已验证的阳性标本为模板检验多重体系的特异性。多重检测体系在103拷贝/μL水平可同时检测到12种病毒。另检测24份临床标本,以real-time RT-PCR为参考标准,进一步验证检测体系。结果表明,这种基于GeXP系统的新方法灵敏度高、特异性强,可以快速同时检测12种常见呼吸道病毒。
Abstract：A GeXP based multiplex RT-PCR assay was developed to simultaneously detect twelve different respiratory viruses types/subtypes including influenza A virus,influenza B virus,influenza A virus sH1N1,parainfluenza virus type 1,parainfluenza virus type 2,parainfluenza virus type 3,human rhinovirus, human metapneumovirus,adenovirus,respiratory syncytial virus A,respiratory syncytial virus B and human bocavirus.Twelve sets of specific primers were designed based on the conserved sequences of available respiratory-virus sequence database The specificity of the multiplex system was examined by positive specimens confirmed previously.The sensitivity to detect twelve respiratory viruses simultaneously was 103 copies/μL.Twenty four clinical specimens were further detected by this novel assay and the results were compared with that of the real-time RT-PCR.These results showed that this novel assay based on GeXP is a fast,sensitive,and high throughput test for the detection of respiratory virus infections.
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基本信息:
DOI：10.13242/j.cnki.bingduxuebao.002218
中图分类号:R440
引用信息:
[1]李瑾,毛乃颖,秦萌,等.GeXP多重PCR技术同时检测12种常见呼吸道病毒[J].病毒学报,2011,27(06):526-532.DOI:10.13242/j.cnki.bingduxuebao.002218.
基金信息:
传染病防治重大专项(2009ZX10004-101,202)
2011-12-30
2011-12-30
2011-12-30