A broad range assay for rapid detection and etiologic characterization of bacterial meningitis: performance testing in samples from sub-Sahara

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• 作者：Helen Won ; Samuel Yang ; Charlotte Gaydos ; Justin Hardick ; Padmini Ramach ; ran ; Yu-Hsiang Hsieh ; Alex ; er Kecojevic ; Berthe-Marie Njanpop-Lafourcade ; Judith E. Mueller ; Tsidi Agbeko Tameklo ; Kossi Badziklou ; Bradford D. Gessner ; Richard E. Rothman
• 关键词：Bacterial meningitis ; Clinical samples ; Meningitis belt ; Real-time PCR ; Disease surveillance
• 刊名：Diagnostic Microbiology and Infectious Disease
• 出版年：2012
• 出版时间：September, 2012
• 年：2012
• 卷：74
• 期：1
• 页码：22-27
• 全文大小：206 K

文摘

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.