GeXP多重PCR技术同时检测12种常见呼吸道病毒

A GeXP based Multiplex RT-PCR Assay for Simultaneous Detection of Twelve Human Respiratory Viruses

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10^3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realtime 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。

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 rhinovi- rus, 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 avail- able 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 103copies/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.