Rapid detection of eight causative pathogens for the diagnosis of bacterial meningitis by real-time PCR

Naoko Chiba, Somay Y. Murayama, Miyuki Morozumi, Eiichi Nakayama, Takafumi Okada, Satoshi Iwata, Keisuke Sunakawa, Kimiko Ubukata

研究成果: Article

62 引用 (Scopus)

抄録

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 (χ2 = 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.

元の言語English
ページ(範囲)92-98
ページ数7
ジャーナルJournal of Infection and Chemotherapy
15
発行部数2
DOI
出版物ステータスPublished - 2009 4
外部発表Yes

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Bacterial Meningitides
Real-Time Polymerase Chain Reaction
Cerebrospinal Fluid
Mycoplasma pneumoniae
Streptococcus agalactiae
Haemophilus influenzae
Listeria monocytogenes
Streptococcus pneumoniae
Escherichia coli
Molecular Probes
Polymerase Chain Reaction
Neisseria meningitidis
Meningitis
Staphylococcus aureus
Limit of Detection
Japan
Newborn Infant
Anti-Bacterial Agents
Drug Therapy
DNA

ASJC Scopus subject areas

  • Microbiology (medical)
  • Pharmacology (medical)
  • Infectious Diseases

これを引用

Rapid detection of eight causative pathogens for the diagnosis of bacterial meningitis by real-time PCR. / Chiba, Naoko; Murayama, Somay Y.; Morozumi, Miyuki; Nakayama, Eiichi; Okada, Takafumi; Iwata, Satoshi; Sunakawa, Keisuke; Ubukata, Kimiko.

:: Journal of Infection and Chemotherapy, 巻 15, 番号 2, 04.2009, p. 92-98.

研究成果: Article

Chiba, N, Murayama, SY, Morozumi, M, Nakayama, E, Okada, T, Iwata, S, Sunakawa, K & Ubukata, K 2009, 'Rapid detection of eight causative pathogens for the diagnosis of bacterial meningitis by real-time PCR', Journal of Infection and Chemotherapy, 巻. 15, 番号 2, pp. 92-98. https://doi.org/10.1007/s10156-009-0670-3
Chiba, Naoko ; Murayama, Somay Y. ; Morozumi, Miyuki ; Nakayama, Eiichi ; Okada, Takafumi ; Iwata, Satoshi ; Sunakawa, Keisuke ; Ubukata, Kimiko. / Rapid detection of eight causative pathogens for the diagnosis of bacterial meningitis by real-time PCR. :: Journal of Infection and Chemotherapy. 2009 ; 巻 15, 番号 2. pp. 92-98.
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abstract = "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 (χ2 = 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.",
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