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Nested PCR Assay for Eight Pathogens: A Rapid Tool for Diagnosis of Bacterial Meningitis

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Abstract

Purpose

Bacterial meningitis is a dreadful infectious disease with a high mortality and morbidity if remained undiagnosed. Traditional diagnostic methods for bacterial meningitis pose a challenge in accurate identification of pathogen, making prognosis difficult. The present study is therefore aimed to design and evaluate a specific and sensitive nested 16S rDNA genus-based polymerase chain reaction (PCR) assay using clinical cerebrospinal fluid (CSF) for rapid diagnosis of eight pathogens causing the disease.

Methods

The present work was dedicated to development of an in-house genus specific 16S rDNA nested PCR covering pathogens of eight genera responsible for causing bacterial meningitis using newly designed as well as literature based primers for respective genus. A total 150 suspected meningitis CSF obtained from the patients admitted to Central India Institute of Medical Sciences (CIIMS), India during the period from August 2011 to May 2014, were used to evaluate clinical sensitivity and clinical specificity of optimized PCR assays.

Results

The analytical sensitivity and specificity of our newly designed genus-specific 16S rDNA PCR were found to be ≥92 %. With such a high sensitivity and specificity, our in-house nested PCR was able to give 100 % sensitivity in clinically confirmed positive cases and 100 % specificity in clinically confirmed negative cases indicating its applicability in clinical diagnosis.

Conclusions

Our in-house nested PCR system therefore can diagnose the accurate pathogen causing bacterial meningitis and therefore be useful in selecting a specific treatment line to minimize morbidity. Results are obtained within 24 h and high sensitivity makes this nested PCR assay a rapid and accurate diagnostic tool compared to traditional culture-based methods.

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Authors and Affiliations

  1. Research Laboratory, Central India Institute of Medical Sciences, Nagpur, Maharashtra, India

    Sharda P. Bhagchandani, Sushant Kubade, Priyanka P. Nikhare, Sonali Manke, Nitin H. Chandak, Dinesh Kabra, Neeraj N. Baheti, Vijay S. Agrawal, Pankaj Sarda, Parikshit Mahajan, Ashish Ganjre, Lokendra Singh, Girdhar M. Taori, Hatim F. Daginawala & Rajpal S. Kashyap

  2. Environmental Genomics Unit, National Environmental Engineering Research Institute, Nagpur, Maharashtra, India

    Hemant J. Purohit

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  1. Sharda P. Bhagchandani
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Corresponding author

Correspondence to Rajpal S. Kashyap.

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Conflict of interest

Sharda P. Bhagchandani, Sushant Kubade, Priyanka P. Nikhare, Sonali Manke, Nitin H. Chandak, Dinesh Kabra, Neeraj N. Baheti, Vijay S. Agrawal, Pankaj Sarda, Parikshit Mahajan, Ashish Ganjre, Hemant J. Purohit, Lokendra Singh, Girdhar M. Taori, Hatim F. Daginawala and Rajpal S. Kashyap declare that they do not have any conflict of interest.

Funding

This is in-house project work funded by Central India Institute of Medical Sciences, Nagpur, Maharashtra, India.

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Bhagchandani, S.P., Kubade, S., Nikhare, P.P. et al. Nested PCR Assay for Eight Pathogens: A Rapid Tool for Diagnosis of Bacterial Meningitis. Mol Diagn Ther 20, 45–54 (2016). https://doi.org/10.1007/s40291-015-0174-z

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