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Research article
Towards accurate exclusion of neonatal bacterial meningitis: a feasibility study of a novel 16S rDNA PCR assay.
Arthur Abelian
Maelor Hospital, Betsi Cadwaladr University Health Board
Corresponding Author
ORCiD: https://orcid.org/0000-0002-9287-6135
License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published version at BMC Infectious Diseases.
Background: PCRctic is an innovative assay based on 16S rDNA PCR technology that has been designed to detect a single intact bacterium in a specimen of cerebro-spinal fluid (CSF). The assay’s potential for accurate, fast and inexpensive discrimination of bacteria-free CSF makes it an ideal adjunct for confident exclusion of bacterial meningitis in newborn babies where the negative predictive value of bacterial culture is poor. This study aimed to stress-test and optimize PCRctic in the “field conditions” to attain a clinically useful level of specificity.
Methods: The specificity of PCRctic was evaluated in CSF obtained from newborn babies investigated for meningitis on a tertiary neonatal unit. Following an interim analysis, the method of skin antisepsis was changed to increase bactericidal effect, and snap-top tubes (EppendorfTM) replaced standard universal containers for collection of CSF to reduce environmental contamination.
Results: The assay’s specificity was 90.5% in CSF collected into the snap-top tubes – up from 60% in CSF in the universal containers. The method of skin antisepsis had no effect on the specificity. All CSF cultures were negative and no clinical cases of neonatal bacterial meningitis occurred during the study.
Conclusions: a simple and inexpensive optimization of CSF collection resulted in a high specificity output. The low prevalence of neonatal bacterial meningitis means that a large multi-centre study will be required to validate the assay’s sensitivity and its negative predictive value.
Keywords
neonatal bacterial meningitis, broad-range PCR, ethidium monoazideneonatal bacterial meningitis, broad-range PCR, ethidium monoazide
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CURRENT STATUS: Published
Version 2
Posted 13 Jun, 2020
No comments provided
Editorial decision: Minor revision
On 09 Jun, 2020
Review #2 received
Received 14 Dec, 2019
Review #1 received
Received 14 Dec, 2019
Reviewer #2 agreed
On 25 Nov, 2019
Editor invited
On 20 Nov, 2019
Reviewers invited
Invitations sent on 20 Nov, 2019
Reviewer #1 agreed
On 20 Nov, 2019
First submitted
On 13 Nov, 2019
Editor assigned
On 13 Nov, 2019
Submission checks complete
On 12 Nov, 2019
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Subject Areas
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This preprint is under consideration at BMC Infectious Diseases. A preprint is a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research article
Towards accurate exclusion of neonatal bacterial meningitis: a feasibility study of a novel 16S rDNA PCR assay.
Arthur Abelian
Maelor Hospital, Betsi Cadwaladr University Health Board
Corresponding Author
ORCiD: https://orcid.org/0000-0002-9287-6135
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Published
Version 2
Posted 13 Jun, 2020
No comments provided
Editorial decision: Minor revision
On 09 Jun, 2020
Review #2 received
Received 14 Dec, 2019
Review #1 received
Received 14 Dec, 2019
Reviewer #2 agreed
On 25 Nov, 2019
Editor invited
On 20 Nov, 2019
Reviewers invited
Invitations sent on 20 Nov, 2019
Reviewer #1 agreed
On 20 Nov, 2019
First submitted
On 13 Nov, 2019
Editor assigned
On 13 Nov, 2019
Submission checks complete
On 12 Nov, 2019
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Infectious Diseases
License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published version at BMC Infectious Diseases.
Background: PCRctic is an innovative assay based on 16S rDNA PCR technology that has been designed to detect a single intact bacterium in a specimen of cerebro-spinal fluid (CSF). The assay’s potential for accurate, fast and inexpensive discrimination of bacteria-free CSF makes it an ideal adjunct for confident exclusion of bacterial meningitis in newborn babies where the negative predictive value of bacterial culture is poor. This study aimed to stress-test and optimize PCRctic in the “field conditions” to attain a clinically useful level of specificity.
Methods: The specificity of PCRctic was evaluated in CSF obtained from newborn babies investigated for meningitis on a tertiary neonatal unit. Following an interim analysis, the method of skin antisepsis was changed to increase bactericidal effect, and snap-top tubes (EppendorfTM) replaced standard universal containers for collection of CSF to reduce environmental contamination.
Results: The assay’s specificity was 90.5% in CSF collected into the snap-top tubes – up from 60% in CSF in the universal containers. The method of skin antisepsis had no effect on the specificity. All CSF cultures were negative and no clinical cases of neonatal bacterial meningitis occurred during the study.
Conclusions: a simple and inexpensive optimization of CSF collection resulted in a high specificity output. The low prevalence of neonatal bacterial meningitis means that a large multi-centre study will be required to validate the assay’s sensitivity and its negative predictive value.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Due to technical limitations, tables are only available as a download in the supplemental files section