PMID- 16000464
OWN - NLM
STAT- MEDLINE
DCOM- 20050826
LR  - 20220311
IS  - 0095-1137 (Print)
IS  - 1098-660X (Electronic)
IS  - 0095-1137 (Linking)
VI  - 43
IP  - 7
DP  - 2005 Jul
TI  - Rapid diagnosis of bacterial meningitis by real-time PCR and fluorescence in situ 
      hybridization.
PG  - 3390-7
AB  - Real-time PCR and fluorescence in situ hybridization (FISH) were evaluated as 
      rapid methods for the diagnosis of bacterial meningitis and compared to standard 
      diagnostic procedures. For PCR, a LightCycler approach was chosen, implementing 
      eubacterial and specific PCR assays for the most relevant bacteria. For FISH, a 
      similar probe set containing eubacterial and specific probes was composed of 
      published and newly designed probes. Both methods were evaluated by use of 
      cerebrospinal fluid (CSF) samples from patients with suspected bacterial 
      meningitis. For all microscopy- and culture-positive samples (n = 28), the 
      eubacterial PCR was positive. In addition, all identifiable pathogens were 
      detected with specific PCR assays, according to an algorithm based on the Gram 
      stain. The FISH method detected the pathogen in 13 of 18 positive samples. While 
      the FISH method remained negative for all microscopy- and culture-negative 
      samples (n = 113), the eubacterial PCR was positive for five of these samples. 
      Sequencing of the amplicon revealed the presence of Neisseria meningitidis, 
      Streptococcus agalactiae, and Haemophilus influenzae in three of these five 
      samples. In addition, samples with discordant results by culture and microscopy 
      were successfully investigated by PCR (10 samples) and FISH (5 samples). In 
      conclusion, PCR is a highly sensitive tool for rapid diagnosis of bacterial 
      meningitis. FISH is less sensitive but is useful for the identification of CSF 
      samples showing bacteria in the Gram stain. Based on our results, an approach for 
      laboratory diagnosis of meningitis including PCR and FISH is discussed.
FAU - Poppert, Sven
AU  - Poppert S
AD  - Department of Medical Microbiology and Hygiene, University of Ulm, Ulm, Germany. 
      sven.poppert@medizin.uni-ulm.de
FAU - Essig, Andreas
AU  - Essig A
FAU - Stoehr, Barbara
AU  - Stoehr B
FAU - Steingruber, Adelinde
AU  - Steingruber A
FAU - Wirths, Beate
AU  - Wirths B
FAU - Juretschko, Stefan
AU  - Juretschko S
FAU - Reischl, Udo
AU  - Reischl U
FAU - Wellinghausen, Nele
AU  - Wellinghausen N
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Clin Microbiol
JT  - Journal of clinical microbiology
JID - 7505564
RN  - 0 (Culture Media)
RN  - 0 (DNA Probes)
RN  - 0 (DNA, Bacterial)
SB  - IM
MH  - Cerebrospinal Fluid/*microbiology
MH  - Culture Media
MH  - DNA Probes
MH  - DNA, Bacterial/analysis/genetics
MH  - Gram-Negative Bacteria/genetics/isolation & purification
MH  - Gram-Positive Cocci/genetics/isolation & purification
MH  - Humans
MH  - *In Situ Hybridization, Fluorescence
MH  - Meningitis, Bacterial/*diagnosis/microbiology
MH  - Microscopy
MH  - Polymerase Chain Reaction/*methods
MH  - Time Factors
PMC - PMC1169125
EDAT- 2005/07/08 09:00
MHDA- 2005/08/27 09:00
PMCR- 2005/07/01
CRDT- 2005/07/08 09:00
PHST- 2005/07/08 09:00 [pubmed]
PHST- 2005/08/27 09:00 [medline]
PHST- 2005/07/08 09:00 [entrez]
PHST- 2005/07/01 00:00 [pmc-release]
AID - 43/7/3390 [pii]
AID - 1291-04 [pii]
AID - 10.1128/JCM.43.7.3390-3397.2005 [doi]
PST - ppublish
SO  - J Clin Microbiol. 2005 Jul;43(7):3390-7. doi: 10.1128/JCM.43.7.3390-3397.2005.