...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Journal of Clinical Microbiology >Prospective Study of Use of PCR Amplification and Sequencing of 16S Ribosomal DNA from Cerebrospinal Fluid for Diagnosis of Bacterial Meningitis in a Clinical Setting
【24h】

Prospective Study of Use of PCR Amplification and Sequencing of 16S Ribosomal DNA from Cerebrospinal Fluid for Diagnosis of Bacterial Meningitis in a Clinical Setting

机译：脑脊液中16S核糖体DNA的PCR扩增和测序在临床诊断细菌性脑膜炎中的应用前瞻性研究

获取原文
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

We have evaluated the use of a broad-range PCR aimed at the 16S rRNA gene in detecting bacterial meningitis in a clinical setting. To achieve a uniform DNA extraction procedure for both gram-positive and gram-negative organisms, a combination of physical disruption (bead beating) and a silica-guanidiniumthiocyanate procedure was used for nucleic acid preparation. To diminish the risk of contamination as much as possible, we chose to amplify almost the entire 16S rRNA gene. The analytical sensitivity of the assay was approximately 1 × 102 to 2 × 102 CFU/ml of cerebrospinal fluid (CSF) for both gram-negative and gram-positive bacteria. In a prospective study of 227 CSF samples, broad-range PCR proved to be superior to conventional methods in detecting bacterial meningitis when antimicrobial therapy had already started. Overall, our assay showed a sensitivity of 86%, a specificity of 97%, a positive predictive value of 80%, and a negative predictive value of 98% compared to culture. We are currently adapting the standard procedures in our laboratory for detecting bacterial meningitis; broad-range 16S ribosomal DNA PCR detection is indicated when antimicrobial therapy has already started at time of lumbar puncture or when cultures remain negative, although the suspicion of bacterial meningitis remains.
机译：我们评估了针对16S rRNA基因的大范围PCR在临床环境中检测细菌性脑膜炎的用途。为了对革兰氏阳性和革兰氏阴性生物均实现统一的DNA提取程序，将物理破坏（微珠敲打）和二氧化硅-胍基硫氰酸盐程序结合使用来制备核酸。为了尽可能降低污染的风险，我们选择扩增几乎整个16S rRNA基因。对于革兰氏阴性菌和革兰氏阳性菌，该分析方法的分析敏感性约为1×10 2 至2×10 2 CFU / ml脑脊液（CSF） 。在对227个CSF样品进行的前瞻性研究中，当抗菌治疗已经开始时，广谱PCR在检测细菌性脑膜炎方面优于传统方法。总体而言，与培养相比，我们的测定显示出86％的敏感性，97％的特异性，80％的阳性预测值和98％的阴性预测值。目前，我们正在采用实验室中用于检测细菌性脑膜炎的标准程序;当仍在怀疑细菌性脑膜炎的时候，在穿刺时或当培养物仍为阴性时，已经开始进行抗菌治疗时，可进行宽范围的16S核糖体DNA PCR检测。

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