A multiplex polymerase chain reaction microarray assay to detect bioterror pathogens in blood
A multiplex polymerase chain reaction microarray assay to detect bioterror pathogens in blood
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DOI：
10.1016/s1525-1578(10)60579-x
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发表时间：
2005-10-01
影响因子：
4.1
通讯作者：
Duncan, R
中科院分区：
文献类型：
--
作者：
Tomioka, K;Peredelchuk, M;Duncan, R
对生物恐怖主义危险的高度关切要求制定措施，确保血液供应的安全。使用血液筛查DNA微阵列对这些试剂进行多重检测是同时筛查许多可疑试剂的敏感且特异的方法。我们已经开发和优化了多重聚合酶链反应微阵列检测筛选血液中的三个潜在的生物恐怖细菌病原体和人类核糖体RNA基因内部控制。经证明，该检测试剂盒的分析灵敏度为50个菌落形成单位/ml，用于检测炭疽芽孢杆菌、土拉热弗朗西丝菌和假结核耶尔森菌（鼠疫耶尔森菌的替代品）。不存在任何假阳性，证明分析特异性高。筛选B。炭疽感染的小鼠血液样品和未感染的对照在临床前应用中证明了有效性和特异性。这项研究证明了微阵列技术的概念，同时筛选血液样本中的多种生物恐怖病原体。
Heightened concern about the dangers of bioterrorism requires that measures be developed to ensure the safety of the blood supply. Multiplex detection of such agents using a blood-screening DNA microarray is a sensitive and specific method to screen simultaneously for a number of suspected agents. We have developed and optimized a multiplex polymerase chain reaction microarray assay to screen blood for three potential bioterror bacterial pathogens and a human ribosomal RNA gene internal control. The analytical sensitivity of the assay was demonstrated to be 50 colony-forming units/ml for Bacillus anthracis, Francisella tularensis, and Yersinia pseudotuberculosis (surrogate for Yersinia pestis). The absence of any false-positives demonstrated high analytical specificity. Screening B. anthracis-infected mouse blood samples and uninfected controls demonstrated effectiveness and specificity in a preclinical application. This study represents proof of the concept of microarray technology to screen simultaneously for multiple bioterror pathogens in blood samples.