Abstract
BackgroundPCRctic 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.MethodsThe 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 (Eppendorf™) replaced standard universal containers for collection of CSF to reduce environmental contamination.ResultsThe 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.ConclusionsA 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.
Highlights
PCRctic is an innovative assay based on 16S rDNA Polymerase chain reaction (PCR) technology that has been designed to detect a single intact bacterium in a specimen of cerebro-spinal fluid (CSF)
With many babies exposed to antibiotics intrapartum [3] and with most babies receiving antibiotics before the lumbar puncture [2], a negative result in CSF bacterial culture
We speculated that an assay sensitive enough to accurately discriminate bacteria-free CSF would facilitate accurate exclusion of bacterial meningitis, with early discontinuation of antibiotic treatment and discharge from hospital
Summary
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). Based on clinical and microbiological data [12,13,14], the rate of neonatal bacterial meningitis in the UK is low (0.2 to 0.3 per 1000 live births). This notwithstanding, Abelian et al BMC Infectious Diseases (2020) 20:441. We developed PCRctic – an assay based on the broad range 16S rDNA PCR technology, enhanced to detect a single bacterium in a standard neonatal CSF specimen (200 μl)
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