Yang Yu, Wang Jing, Wen Haiyan, Liu Hengchuan
Institute of Health Quarantine, Chinese Academy of Inspection and Quarantine, Beijing 100123, China.
J Biomed Biotechnol. 2012;2012:831052. doi: 10.1155/2012/831052. Epub 2012 May 29.
We have developed novel Bio-Plex assays for simultaneous detection of Bacillus anthracis, Yersinia pestis, Brucella spp., Francisella tularensis, and Burkholderia pseudomallei. Universal primers were used to amplify highly conserved region located within the 16S rRNA amplicon, followed by hybridized to pathogen-specific probes for identification of these five organisms. The other assay is based on multiplex PCR to simultaneously amplify five species-specific pathogen identification-targeted regions unique to individual pathogen. Both of the two arrays are validated to be flexible and sensitive for simultaneous detection of bioterrorism bacteria. However, universal primer PCR-based array could not identify Bacillus anthracis, Yersinia pestis, and Brucella spp. at the species level because of the high conservation of 16S rDNA of the same genus. The two suspension arrays can be utilized to detect Bacillus anthracis sterne spore and Yersinia pestis EV76 from mimic "write powder" samples, they also proved that the suspension array system will be valuable tools for diagnosis of bacterial biothreat agents in environmental samples.
我们开发了新型生物芯片检测方法，用于同时检测炭疽芽孢杆菌、鼠疫耶尔森菌、布鲁氏菌属、土拉弗朗西斯菌和类鼻疽伯克霍尔德菌。使用通用引物扩增位于16S rRNA扩增子内的高度保守区域，随后与病原体特异性探针杂交以鉴定这五种生物。另一种检测方法基于多重PCR，可同时扩增单个病原体特有的五个物种特异性病原体鉴定靶向区域。这两种检测方法均经验证，对于同时检测生物恐怖主义细菌具有灵活性和敏感性。然而，基于通用引物PCR的芯片检测方法无法在物种水平上鉴定炭疽芽孢杆菌、鼠疫耶尔森菌和布鲁氏菌属，因为同一属的16S rDNA具有高度保守性。这两种悬浮芯片可用于从模拟“炭疽粉末”样本中检测炭疽芽孢杆菌斯特恩芽孢和鼠疫耶尔森菌EV76，它们还证明了悬浮芯片系统将成为环境样本中细菌生物威胁因子诊断的有价值工具。
