DNA探针快速检测肺炎链球菌和流感嗜血杆菌

Hui-zhen Fan, Hua-peng Yu, Wen-jie Huang
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引用次数: 0

摘要

目的:建立肺炎链球菌和流感嗜血杆菌感染早期快速分子检测方法。方法:采用聚合酶链式反应(PCR)合成肺炎链球菌和流感嗜血杆菌特异性DNA探针和细菌16s rRNA通用探针,并用生物素标记。将细菌、病毒和真菌的DNA分别与这些探针杂交,然后应用于临床样品的检测。结果:PCR扩增出263,351和370 bp的DNA探针。肺炎链球菌和流感嗜血杆菌仅与其对应的探针发生反应,细菌通用探针未与病毒和真菌发生交叉反应。该方法可以检测到小至1ng的细菌DNA。在100份痰样本中,肺炎链球菌阳性11份,流感嗜血杆菌阳性8份,阳性率高于痰培养。结论:DNA探针检测肺炎链球菌和流感嗜血杆菌具有简便、快速、特异性强的特点。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
[Rapid detection of Streptococcus pneumoniae and Haemophilus influenzae with DNA probes].

Objective: To establish a method for rapid molecular detection of Streptococcus pneumoniae and Haemophilus influenzae in the early stage of infection.

Methods: Specific DNA probes for Streptococcus pneumoniae and Haemophilus influenzae and 16 S rRNA universal probe of bacteria were synthesized by polymerase chain reaction (PCR) and labeled with biotin. The DNA of the bacteria, virus and fungi were hybridized with these probes respectively prior to application for examination of the clinical samples.

Results: The DNA probes of 263, 351, and 370 bp were amplified by PCR. Streptococcus pneumoniae and Haemophilus influenzae reacted only with their corresponding probes, and no cross reaction of the bacterial universal probe with virus and fungi was noted. The method could detect bacterial DNA in as small amount as 1 ng. Of the 100 sputum specimens, 11 were found to be positive for Streptococcus pneumoniae and 8 for Haemophilus influenzae, with a positivity rate greater than that by sputum culture.

Conclusion: DNA probe detection is simple, rapid, and specific for clinical examination of Streptococcus pneumoniae and Haemophilus influenzae.

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