...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Journal of Clinical Microbiology >Specific detection of Mycobacterium tuberculosis complex strains by polymerase chain reaction.
【24h】

Specific detection of Mycobacterium tuberculosis complex strains by polymerase chain reaction.

机译：通过聚合酶链反应特异性检测结核分枝杆菌复杂菌株。

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

摘要

During the screening of a Mycobacterium tuberculosis lambda gt-11 gene library with monoclonal antibodies, we detected a recombinant clone, lambda PH7311, which contained a mycobacterial DNA insert that hybridized specifically with DNA of M. tuberculosis complex strains. Part of this insert was sequenced and used for the development of an M. tuberculosis complex-specific polymerase chain reaction (PCR). Only strains belonging to species of the M. tuberculosis complex group contained an amplifiable fragment of 158 base pairs (bp). This fragment was absent in all strains tested belonging to 15 other mycobacterial species. After amplification by PCR and dot blot hybridization with a digoxigenin-labeled oligonucleotide, the limit of detection of purified genomic M. tuberculosis DNA amounted to a quantity corresponding to 20 bacterial cells. By this technique about 10(3) M. tuberculosis bacteria were detectable in sputum. Using PCR, we were also able to detect M. tuberculosis cells in clinical material such as pleural fluid, bronchial washings, and biopsies, and these results were comparable with those obtained by classical bacterial culture. Of 34 M. tuberculosis strains, 5 did not carry the amplifiable 158-bp fragment, which occurs usually as a single copy in the chromosome. Evidence is presented that the 158-bp fragment is located near a repeated sequence in the chromosome. We presume that strains which did not carry the 158-bp fragment have lost a chromosomal segment by a genetic rearrangement induced by the repetitive DNA element.
机译：在用单克隆抗体筛选结核分枝杆菌λgt-11基因文库的过程中，我们检测到一个重组克隆λPH7311，其中含有一个与结核分枝杆菌复杂菌株的DNA特异性杂交的分枝杆菌DNA插入片段。对该插入物的一部分进行测序，并用于开发结核分枝杆菌复合物特异性聚合酶链反应（PCR）。仅属于结核分枝杆菌复杂群物种的菌株包含158个碱基对（bp）的可扩增片段。在属于15个其他分枝杆菌物种的所有测试菌株中均没有此片段。通过PCR扩增并与洋地黄毒苷标记的寡核苷酸进行斑点印迹杂交后，纯化的基因组结核分枝杆菌DNA的检出限为相当于20个细菌细胞的量。通过该技术，在痰中可检测到约10（3）个结核分枝杆菌。使用PCR，我们还能够在临床材料（例如胸膜液，支气管冲洗液和活检组织）中检测结核分枝杆菌细胞，这些结果与经典细菌培养所获得的结果相当。在34个结核分枝杆菌菌株中，有5个未携带可扩增的158 bp片段，该片段通常以单拷贝形式出现在染色体上。证据表明158 bp的片段位于染色体的重复序列附近。我们推测不携带158bp片段的菌株由于重复DNA元件引起的基因重排而丢失了染色体片段。

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