Li Jin, Mao Nai-Ying, Qin Meng, Hu Xiu-Mei, Yang Meng-Jie, Wang Miao, Zhang Chen, Xu Wen-Bo, Ma Xue-Jun
State Key Laboratory for Genetic Engineering and Molecular Virology, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China.
Bing Du Xue Bao. 2011 Nov;27(6):526-32.
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 10(3) copies/microL. 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.
开发了一种基于GeXP的多重逆转录聚合酶链反应(RT-PCR)检测方法,用于同时检测12种不同的呼吸道病毒类型/亚型,包括甲型流感病毒、乙型流感病毒、甲型H1N1流感病毒、1型副流感病毒、2型副流感病毒、3型副流感病毒、人鼻病毒、人偏肺病毒、腺病毒、呼吸道合胞病毒A、呼吸道合胞病毒B和人博卡病毒。基于现有呼吸道病毒序列数据库的保守序列设计了12组特异性引物。通过先前确认的阳性标本检测多重系统的特异性。同时检测12种呼吸道病毒的灵敏度为10(3)拷贝/微升。用这种新方法进一步检测了24份临床标本,并将结果与实时RT-PCR的结果进行比较。这些结果表明,这种基于GeXP的新方法是一种用于检测呼吸道病毒感染的快速、灵敏且高通量的检测方法。