A broad range assay for rapid detection and etiologic characterization of bacterial meningitis: performance testing in samples from sub-Sahara.
This study aimed to conduct a pilot evaluation of broad-based multiprobe polymerase chain reaction (PCR) in clinical cerebrospinal fluid (CSF) samples compared to local conventional PCR/culture methods used for bacterial meningitis surveillance. A previously described PCR consisting of initial broad-based detection of Eubacteriales by a universal probe, followed by Gram typing, and pathogen-specific probes was designed targeting variable regions of the 16S rRNA gene. The diagnostic performance of the 16S rRNA assay in ""127 CSF samples was evaluated in samples from patients from Togo, Africa, by comparison to conventional PCR/culture methods. Our probes detected Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae. Uniprobe sensitivity and specificity versus conventional PCR were 100% and 54.6%, respectively. Sensitivity and specificity of uniprobe versus culture methods were 96.5% and 52.5%, respectively. Gram-typing probes correctly typed 98.8% (82/83) and pathogen-specific probes identified 96.4% (80/83) of the positives. This broad-based PCR algorithm successfully detected and provided species level information for multiple bacterial meningitis agents in clinical samples.
Department of Emergency Medicine, Johns Hopkins University, Baltimore, MD, USA.
Helen
H
Won
Samuel
S
Yang
Charlotte
C
Gaydos
Justin
J
Hardick
Padmini
P
Ramachandran
Yu-Hsiang
YH
Hsieh
Alexander
A
Kecojevic
Berthe-Marie
BM
Njanpop-Lafourcade
Judith E
JE
Mueller
Tsidi Agbeko
TA
Tameklo
Kossi
K
Badziklou
Bradford D
BD
Gessner
Richard E
RE
Rothman
D001431:Bacteriological Techniques / Q000379:methods*; D002555:Cerebrospinal Fluid / Q000382:microbiology; D004269:DNA, Bacterial / Q000235:genetics; D004275:DNA, Ribosomal / Q000235:genetics; D006193:Haemophilus influenzae / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D008583:Meningitis, Haemophilus / Q000175:diagnosis* / Q000382:microbiology; D008585:Meningitis, Meningococcal / Q000175:diagnosis* / Q000382:microbiology; D008586:Meningitis, Pneumococcal / Q000175:diagnosis* / Q000382:microbiology; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D009345:Neisseria meningitidis / Q000235:genetics / Q000302:isolation & purification; D015345:Oligonucleotide Probes / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012680:Sensitivity and Specificity; D013296:Streptococcus pneumoniae / Q000235:genetics / Q000302:isolation & purification; D013997:Time Factors; D014037:Togo
D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D015345:Oligonucleotide Probes; D012336:RNA, Ribosomal, 16S