One‐step heminested PCR for amplification of Neisseria meningitidis DNA in cerebrospinal fluid

摘要

A one‐step polymerase chain reaction (Heminested‐PCR) was designed to target the 16S rRNA fragment simultaneously using a set of primers for the universal bacterial group and a species‐specific sequence for diagnostic purposes. The diagnostic features of the Heminested‐PCR were evaluated in the study of 168 cerebrospinal fluid (CSF) specimens from 84 patients with a infection, meningitis caused by unrelated bacteria and other etiologies (57 patients), or suspicious cases (27 patients) with clinical symptoms of bacterial meningitis but with negative results from bacteriological procedures. About 90% of patients with bacterial meningitis, including those suspicious cases, had prior antibiotic therapy. The sensitivity, specificity, positive, and negative predictive values found in relation to culture and/or microscopy were 91.7, 100, 100, 100, and 90.5%, respectively. In patients suspected of having bacterial meningitis, the Heminested‐PCR revealed 51.9% (14 patients) positive for infection and 40.7% (11 patients) positive for unrelated bacterial infections. The agreement of the Heminested‐PCR with culture and/or microscopy was high and ranked as (kappa indices > 0.856), in contrast to its agreement with other techniques. These findings speak in favor of the molecular diagnosis of meningococcal meningitis in patients who are culture‐ and/or microscopy‐negative, due to their prior antibiotic treatment. J. Clin. Lab. Anal. 14:193–199, 2000. © 2000 Wiley‐Liss, Inc.