Rapid detection of eight causative pathogens for the diagnosis of bacterial meningitis by real-time PCR.
We aimed to detect causative pathogens in cerebrospinal fluid (CSF) collected from patients diagnosed with bacterial meningitis by real-time polymerase chain reaction (PCR). In addition to Streptococcus pneumoniae, Haemophilus influenzae, and Mycoplasma pneumoniae described previously, five other pathogens, Neisseria meningitidis, Escherichia coli, Streptococcus agalactiae, Staphylococcus aureus, and Listeria monocytogenes, were targeted, based on a large-scale surveillance in Japan. Results in CSF from neonates and children (n=150), and from adults (n=18) analyzed by real-time PCR with molecular beacon probes were compared with those of conventional culturing. The total time from DNA extraction from CSF to PCR analysis was 1.5 h. The limit of detection for these pathogens ranged from 5 copies to 28 copies per tube. Nonspecific positive reactions were not recognized for 37 microorganisms in clinical isolates as a negative control. The pathogens were detected in 72.0% of the samples by real-time PCR, but in only 48.2% by culture, although the microorganisms were completely concordant. With the real-time PCR, the detection rate of H. influenzae from CSF was high, at 45.2%, followed by S. pneumoniae (21.4%), S. agalactiae (2.4%), E. coli (1.8%), L. monocytogenes (0.6%), and M. pneumoniae (0.6%). The detection rate with PCR was significantly better than that with cultures in patients with antibiotic administration (chi2=18.3182; P=0.0000). In conclusion, detection with real-time PCR is useful for rapidly identifying the causative pathogens of meningitis and for examining the clinical course of chemotherapy.
Chiba
Naoko
N
Laboratory of Molecular Epidemiology for Infectious Agents, Graduate School of Infection Control Sciences, Kitasato University, 5-9-1 Shirokane, Minato-ku, Tokyo, 108-8641, Japan.
Murayama
Somay Y
SY
Morozumi
Miyuki
M
Nakayama
Eiichi
E
Okada
Takafumi
T
Iwata
Satoshi
S
Sunakawa
Keisuke
K
Ubukata
Kimiko
K
D000900:Anti-Bacterial Agents / Q000008:administration & dosage; D015373:Bacterial Typing Techniques / Q000379:methods*; D002555:Cerebrospinal Fluid / Q000382:microbiology; D016009:Chi-Square Distribution; D006090:Gram-Negative Bacteria / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification; D006094:Gram-Positive Bacteria / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification; D006193:Haemophilus influenzae / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D016920:Meningitis, Bacterial / Q000175:diagnosis* / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013296:Streptococcus pneumoniae / Q000187:drug effects / Q000235:genetics / Q000302:isolation & purification
D000900:Anti-Bacterial Agents