...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Molecular and Cellular Probes: The Location, Diagnosis and Monitoring of Disease by Specific Molecules and Cell Lines >Rapid and sensitive PCR-based detection and differentiation of aetiologic agents of human granulocytotropic and monocytotropic ehrlichiosis.
【24h】

Rapid and sensitive PCR-based detection and differentiation of aetiologic agents of human granulocytotropic and monocytotropic ehrlichiosis.

机译：基于快速和敏感的基于PCR的人类粒细胞性和单细胞性埃希氏病病原学检测和区分。

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 potential of fatal outcome for patients afflicted with human ehrlichioses (HME and HGE) necessitates fast and accurate detection of the aetiologic agents and timely antibiotic treatment. A polymerase chain reaction (PCR)-based protocol is described that can detect as little as 10 copies of ehrlichial 16S rDNA and as few as 0.3 HGE-infected neutrophils. The method employs DNAzol for rapid DNA extraction from unfractionated whole blood in less than 1 h. For DNA amplification, highly specific oligonucleotide primers are designed that efficiently detect and distinguish between Ehrlichia chaffeensis and the HGE agent. These primers do not prime DNA extracted from closely related ehrlichial and rickettsial species. Although total DNA extracted from human blood contains nucleic acids that can be non-specifically amplified at moderate to high MgCI2 concentrations, such non-specific priming of non-ehrlichial DNA can be completely eliminated by lowering the MgCI2 concentration to 1 mM. Thus, this PCR-based procedure can detect and differentiate HGE and HME with speed, simplicity, specificity and sensitivity.
机译：对于遭受人类水杨糖（HME和HGE）折磨的患者而言，致命的潜在结果需要迅速，准确地检测出病因并及时进行抗生素治疗。描述了一种基于聚合酶链反应（PCR）的方案，该方案可检测少至10个拷贝的埃希氏16S rDNA和少至0.3个感染HGE的中性粒细胞。该方法使用DNAzol在不到1小时的时间内从未分离的全血中快速提取DNA。对于DNA扩增，设计了高度特异性的寡核苷酸引物，可有效检测和区分恰菲埃里希氏菌和HGE试剂。这些引物不会引发从密切相关的埃希氏菌属和立克次氏菌属物种中提取的DNA。尽管从人血中提取的总DNA包含可以在中等至高MgCl2浓度下非特异性扩增的核酸，但通过将MgCl2浓度降低至1 mM，可以完全消除这种非水合DNA的非特异性引发。因此，这种基于PCR的程序可以快速，简单，特异性和灵敏地检测和区分HGE和HME。

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