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Clinical use of multiplex-PCR for the diagnosis of acute bacterial meningitis
Open accessAbstract
Background and Objectives: Prompt and accurate diagnosis of acute bacterial meningitis (ABM) is critical for patient management. We designed and evaluated two sets of multiplex-PCR assays for the simultaneous detection of six major etiologies of ABM i.e., Streptococcus pneumoniae, Haemophilus influenzae type b, and Neisseria meningitidis in one set and Listeria monocytogenes, Streptococcus agalactiae, and Escherichia coli in another set of multiplex-PCR in CSF of patients with suspected ABM. Methods: A total of 113 CSF specimens from patients of all ages having clinical features suggestive of meningitis were tested for bacteriological evidence by Gram’s smear, culture, and our designed multiplex-PCR. Results: Multiplex-PCR assay performed excellently by increasing the overall detection rate by 6% when compared to culture as of total 113 samples tested, 17 (15%) were positive by multiplex-PCR whereas only 9% (10/113) were positive by culture. It detected the DNA in eight culture negative samples revealing the presence of S. pneumoniae in three and other possible bacterial pathogens in five of them. Our assay showed a DNA detection limit of 1 pg/μL. Compared to CSF culture, the sensitivity and specificity of the multiplex-PCR were 90% and 92.2%, respectively. Conclusion: This study accentuates the importance of multiplex-PCR assay that is efficiently fast and reliable for the diagnosis of acute bacterial meningitis that can substantially improve the diagnosis in culture negative cases, especially in patients who were previously started on antimicrobial therapy.
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“Major challenges remain, however, in the deployment of these assays in low- and middle-income countries, due to the variability of laboratory capabilities, shortage of trained laboratory personnel and challenges in procurement of reagents and equipment [9, 10-11].”
From Herpetic viral dominance in pediatric meningitis and meningoencephalitis (2026) · cites this paper as [11]
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Epidemiology, Diagnosis, and Antimicrobial Treatment of Acute Bacterial Meningitis
Brouwer, M. C., Tunkel, A. R., & van de Beek, D. (2010) · Clinical Microbiology Reviews · doi:10.1128/cmr.00070-09 · Cited by 930 · cited 1×
“[9,10] The sensitivity of CSF culture, the gold standard method for its microbiological confirmation is around 85% in patients without antibiotic therapy prior to sample collection[1,11,12] with 30% more reduction under antibiotic therapy,[13] which limits its utility.”
Acute bacterial meningitis in infants and children
Kim, K. S. (2010) · The Lancet Infectious Diseases · doi:10.1016/s1473-3099(09)70306-8 · Cited by 454
Choi, J. J., Westblade, L. F., Gottesdiener, L. S., Liang, K., Li, H. A., Wehmeyer, G. T., Glesby, M. J., & Simon, M. S. (2021) · Open Forum Infectious Diseases · doi:10.1093/ofid/ofab467 · Cited by 21 · cited 1×
“The implementation of such tests for ABM has been reported to in fact reduce the duration of empiric antimicrobial therapy and possibly the length of hospital stay when compared with conventional diagnostic methods.[29]”
Result
Research works printed but not linked: 24
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Cited by
Sorted by citations
Herpetic viral dominance in pediatric meningitis and meningoencephalitis
Mishra, P., Pandey, S. C., Shukla, S. K., Sharma, A., Singh, S. P., Jais, M. B., Tiwari, S., Gogoi, S., & Yadav, V. (2026) · Bioinformation · doi:10.6026/973206300221448
Metadata
- Authors
- Sharma, N., Gautam, H., Tyagi, S., Raza, S., Mohapatra, S., Sood, S., Dhawan, B., Kapil, A., & Das, B. K.
- Affiliations
- All India Institute of Medical Sciences (IN)
- Department of Microbiology, All India Institute of Medical Sciences, New Delhi, India
- Corresponding author
- Gautam, Hitender
- Venue
- Journal of Family Medicine and Primary Care
- Published
- 2022-02
- Volume
- 11
- Issue
- 2
- Pages
- 593-598
- ISSN
- 2249-4863 (ISSN-L)
- 2278-7135
- PMC
- PMC8963590
- OpenAlex
- W4214708925
- Cited by
- 21
- References
- 3
- Citations by year
- 2026: 6 · 2025: 6 · 2024: 2 · 2023: 7
- FWCI
- 2.34
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- cc-by-nc-sa
- Locations
- Journal of Family Medicine and Primary Care (publishedVersion, cc-by-nc-sa)
- PubMed Central (publishedVersion, cc-by-nc-sa)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Topics
- Life Sciences › Immunology and Microbiology › Microbiology › Bacterial Infections and Vaccines
- Health Sciences › Medicine › Microbiology (medical) › Bacterial Identification and Susceptibility Testing
- Health Sciences › Medicine › Infectious Diseases › Infectious Diseases and Tuberculosis
- Keywords
- acute bacterial meningitis, multiplex PCR, cerebrospinal fluid, Streptococcus pneumoniae, Haemophilus influenzae type b, Neisseria meningitidis, Listeria monocytogenes, Streptococcus agalactiae, Escherichia coli, culture-negative meningitis, molecular diagnosis, sensitivity and specificity
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