TY - JOUR AU - Diawara, Idrissa AU - Katfy, Khalid AU - Zerouali, Khalid AU - Belabbes, Houria AU - Elmdaghri, Naima PY - 2016/01/31 Y2 - 2024/03/29 TI - A duplex real-time PCR for the detection of Streptococcus pneumoniae and Neisseria meningitidis in cerebrospinal fluid JF - The Journal of Infection in Developing Countries JA - J Infect Dev Ctries VL - 10 IS - 01 SE - Original Articles DO - 10.3855/jidc.5647 UR - https://jidc.org/index.php/journal/article/view/26829537 SP - 53-61 AB - <p class="SmallText">Introduction: Acute bacterial meningitis is one of the most severe infectious diseases. Rapid, accurate, and inexpensive diagnosis of bacterial meningitis is crucial for patient management. This study describes a duplex real-time (RT) PCR assay for detection of <em>Neisseria meningitidis </em>and <em>Streptococcus pneumoniae</em> in the cerebrospinal fluid (CSF) for meningitis diagnosis using SYBR Green-based RT-PCR method coupled with melting curve analysis.</p> <p class="SmallText">Methodology: We used SYBR Green-based RT-PCR method coupled with melting curve analysis to detect <em>S. pneumoniae</em> and <em>N. meningitidis</em> in CSF samples. The sensitivity, specificity, and limit of detection were determined. The gold standard for routine tests of CSF analysis is direct examination, culture, and/or latex agglutination. The assay was evaluated on 132 CSF samples to measure clinical sensitivity.</p> <p class="SmallText">Results: A duplex RT-PCR assay for <em>N. meningitidis </em>and <em>S. pneumoniae</em> detection in CSF was evaluated. Two peaks at different melting temperatures (87.5°C and 85.5°C) for <em>N. meningitidis </em>and<em> S. pneumoniae</em>,<em> </em>respectively, were obtained. The sensitivity of RT-PCR was 100% (95% confidence limits [CI] = 82.4–100) for <em>N. meningitidis </em>and 100% (95% CI = 85.1–100) for <em>S. pneumoniae</em>. Specificity was the same (100%) for the bacteria (95% CI = 88.6–100). The percentage of cases accurately diagnosed with meningitis caused by <em>N. meningitidis </em>and <em>S. pneumoniae</em> increased to 50.7% and 28.6%, respectively, when RT-PCR was added to the standard microbiologic methods.</p> <p class="SmallText">Conclusions: Duplex RT-PCR and melting curve analysis with SYBR Green is an inexpensive, sensitive, and specific method to rapidly diagnose bacterial meningitis. Accurate identification of the bacterial causative agents will improve patient management and epidemiological investigations.</p> ER -