Kotilainen P, Jalava J, Meurman O, Lehtonen O P, Rintala E, Seppälä O P, Eerola E, Nikkari S
Department of Medicine, Turku University Central Hospital, Finland.
J Clin Microbiol. 1998 Aug;36(8):2205-9. doi: 10.1128/JCM.36.8.2205-2209.1998.
We used broad-range bacterial PCR combined with DNA sequencing to examine prospectively cerebrospinal fluid (CSF) samples from patients with suspected meningitis. Fifty-six CSF samples from 46 patients were studied during the year 1995. Genes coding for bacterial 16S and/or 23S rRNA genes could be amplified from the CSF samples from five patients with a clinical picture consistent with acute bacterial meningitis. For these patients, the sequenced PCR product shared 98.3 to 100% homology with the Neisseria meningitidis sequence. For one patient, the diagnosis was initially made by PCR alone. Of the remaining 51 CSF samples, for 50 (98.0%) samples the negative PCR findings were in accordance with the negative findings by bacterial culture and Gram staining, as well as with the eventual clinical diagnosis for the patient. However, the PCR test failed to detect the bacterial rRNA gene in one CSF sample, the culture of which yielded Listeria monocytogenes. These results invite new research efforts to be focused on the application of PCR with broad-range bacterial primers to improve the etiologic diagnosis of bacterial meningitis. In a clinical setting, Gram staining and bacterial culture still remain the cornerstones of diagnosis.
我们采用广谱细菌PCR结合DNA测序技术,对疑似脑膜炎患者的脑脊液(CSF)样本进行前瞻性检测。1995年期间,对46例患者的56份脑脊液样本进行了研究。编码细菌16S和/或23S rRNA基因的基因可从5例临床表现符合急性细菌性脑膜炎的患者的脑脊液样本中扩增出来。对于这些患者,测序的PCR产物与脑膜炎奈瑟菌序列具有98.3%至100%的同源性。对于1例患者,最初仅通过PCR做出诊断。在其余51份脑脊液样本中,50份(98.0%)样本的PCR阴性结果与细菌培养、革兰氏染色的阴性结果以及患者最终的临床诊断一致。然而,PCR检测未能在1份脑脊液样本中检测到细菌rRNA基因,而该样本培养出了产单核细胞李斯特菌。这些结果促使新的研究工作聚焦于应用带有广谱细菌引物的PCR技术,以改善细菌性脑膜炎的病因诊断。在临床环境中,革兰氏染色和细菌培养仍然是诊断的基石。