• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Forensic science international >Multi-platform comparison of ten commercial master mixes for probe-based real-time polymerase chain reaction detection of bioterrorism threat agents for surge preparedness
【24h】

Multi-platform comparison of ten commercial master mixes for probe-based real-time polymerase chain reaction detection of bioterrorism threat agents for surge preparedness

机译：十种商业预混液的多平台比较，用于基于探针的实时检测生物恐怖威胁因子的聚合酶链反应，以进行浪涌防范

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

The Centers for Disease Control and Prevention and United States Army Research Institute for Infectious Diseases have developed real-time PCR assays for the detection of bioterrorism threat agents. These assays all rely on a limited number of approved real-time PCR master mixes. Because the availability of these reagents is a critical element of bioterrorism preparedness, we undertook a joint national preparedness exercise to address the potential surge needs resulting from a large-scale bio-emergency. We identified 9 commercially-available potential alternatives to an existing approved master mix (LightCycler FastStart DNA Master HybProbes): the TaqMan Fast Universal PCR master mix, OmniMix HS, FAST qPCR master mix, EXPRESS qPCR SuperMix kit, QuantiFast Probe PCR kit, LightCycler FastStart DNA MasterPLUS HybProbe, Brilliant II FAST qPCR master mix, ABsolute Fast QPCR Mix and the HotStart IT Taq master mix. The performances of these kits were evaluated by the use of real-time PCR assays for four bioterrorism threat agents: Bacillus anthracis, Brucella melitensis, Burkholderia mallei and Francisella tularensis. The master mixes were compared for target-specific detection levels, as well as consistency of results among three different real-time PCR platforms (LightCycler, SmartCycler and 7500 Fast Dx). Real-time PCR analysis revealed that all ten kits performed well for agent detection on the 7500 Fast Dx instrument; however, the QuantiFast Probe PCR kit yielded the most consistently positive results across multiple real-time PCR platforms. We report that certain combinations of commonly used master mixes and instruments are not as reliable as others at detecting low concentrations of target DNA. Furthermore, our study provides laboratories the option to select from the commercial kits we evaluated to suit their preparedness needs.
机译：疾病控制与预防中心和美国陆军传染病研究所已经开发了用于检测生物恐怖主义威胁因素的实时PCR分析方法。这些测定均依赖于有限数量的批准的实时PCR预混液。由于这些试剂的可用性是生物恐怖主义防备的关键因素，因此我们进行了联合国家防备演习，以解决大规模生物紧急情况造成的潜在紧急需求。我们确定了9种商用替代现有认可的预混液（LightCycler FastStart DNA Master HybProbes）的潜在替代品：TaqMan快速通用PCR预混液，OmniMix HS，FAST qPCR预混液，EXPRESS qPCR SuperMix试剂盒，QuantiFast Probe PCR试剂盒，LightCycler FastStart DNA MasterPLUS HybProbe，Brilliant II FAST qPCR预混液，ABsolute Fast QPCR预混液和HotStart IT Taq预混液。这些试剂盒的性能通过实时荧光定量PCR检测四种生物恐怖威胁因素进行评估：炭疽芽孢杆菌，布鲁氏菌，马尔克伯克霍尔德氏菌和土拉弗朗西斯菌。比较了主要混合物的靶标特异性检测水平以及三种不同的实时PCR平台（LightCycler，SmartCycler和7500 Fast Dx）之间的结果一致性。实时PCR分析显示，所有十种试剂盒在7500 Fast Dx仪器上进行试剂检测均表现良好。但是，QuantiFast探针PCR试剂盒在多个实时PCR平台上产生最一致的阳性结果。我们报告说，常用的主要混合物和仪器​​的某些组合在检测低浓度目标DNA方面不如其他组合可靠。此外，我们的研究为实验室提供了从我们评估的商业工具包中进行选择的选项，以适应他们的备灾需求。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号