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Diagnosis of Meningococcal Infection Using Internally Controlled Multiplex Real-Time PCR

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Neisseria meningitidis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1969))

Abstract

Neisseria meningitidis (Nm) is a leading cause of invasive infections associated with high mortality and morbidity, notably meningitis and septicemia. Etiological rapid diagnosis is key for the preventive management of invasive meningococcal disease (IMD). However, conventional methods for diagnosis are time-consuming and could be hampered by the difficulties in culturing the isolates from clinical specimens especially due to early antibiotic treatment. Therefore, sensitive, specific and rapid non-culture-based methods are valuable for early diagnosis, effective therapy, and prevention. Here we describe a real-time PCR multiplex assays for the detection of Nm targeting the meningococcal-specific gene crgA, coding for a LysR-like transcriptional regulator, and six serogroup-specific (A, B, C, W, X, Y) Nm capsular genes, using a Qiagen column-based method for the optimum isolation of DNA from clinical specimens. Internal quality controls were included to monitor extraction of DNA, inhibition and the technical validation of the PCR as well.

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Acknowledgments

This work was supported by the Institut Pasteur, Paris.

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Authors and Affiliations

  1. Invasive Bacterial Infections Unit, Institut Pasteur, Paris Cedex, France

    Ala-Eddine Deghmane, Eva Hong & Muhamed-Kheir Taha

Authors
  1. Ala-Eddine Deghmane
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  2. Eva Hong
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  3. Muhamed-Kheir Taha
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Corresponding author

Correspondence to Ala-Eddine Deghmane .

Editor information

Editors and Affiliations

  1. Institute For Glycomics, Griffith University, Southport, QLD, Australia

    Kate L. Seib

  2. Institute For Glycomics, Griffith University, Southport, QLD, Australia

    Ian R. Peak

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Deghmane, AE., Hong, E., Taha, MK. (2019). Diagnosis of Meningococcal Infection Using Internally Controlled Multiplex Real-Time PCR. In: Seib, K., Peak, I. (eds) Neisseria meningitidis. Methods in Molecular Biology, vol 1969. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9202-7_2

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  • DOI: https://doi.org/10.1007/978-1-4939-9202-7_2

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9201-0

  • Online ISBN: 978-1-4939-9202-7

  • eBook Packages: Springer Protocols

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