Abstract

Share on facebook
Share on twitter
Share on linkedin

Towards high accuracy meningitis testing in neonates

Presented as a poster at the Neonatal Society 2016 Summer Meeting.

Abelian A1, Mund T2, Pelham H2, Ogilvy-Stuart A3, Curran M4, Dear PH5

1 Maelor Hospital, Wrexham, UK
2 Laboratory of Molecular Biology, MRC Centre, Cambridge, UK
3 Rosie Maternity Hospital, Cambridge, UK
4 Molecular Diagnostics Laboratory, Addenbrooke’s Hospital, Cambridge, UK
5 Mote Research Ltd., Cambridge, UK

Background: The negative predictive value of bacterial culture is low and often inadequate to confidently rule out neonatal bacterial meningitis (NBM). This results in over-testing and over-treatment with prolonged hospitalisation and antibiotic use. Molecular techniques have the potential to augment and even replace bacterial culture.

Methods: “PCRctic” – novel assay based on 16S rDNA PCR technology – utilises ethidium monoazide to eliminate false positive results due to dead bacteria and/or contaminating bacterial DNA, and a single-step closed-tube nested PCR format to achieve sensitivity of single bacterium per 200 microlitres of CSF (5 drops). It offers fast turn-around time (under 2 hours) and minimal per assay cost (under £1). In-built internal control enables to identify false-negative results. It has been optimised to work on the platform of real-time PCR equipment – widely available in NHS molecular diagnostics laboratories.

Results: “PCRctic” detected a single colony-forming unit of the common causes of NBM: E coli, Streptococcus agalactiae (Group B Streptococcus), Streptococcus pneumoniae, and Listeria monocytogenes, whilst avoiding false positive results from contaminating bacterial DNA. Sequence analysis of the rarer bacteria implicated in NBM – Serratia marcescens, Staphylococcus aureus, Salmonella berta and agama, Ureaplasma parvum, Pasteurella multocida, Enterobacter sakazakii (Cronobacter), Sneathia species, Chryseobacterium meningosepticum – showed full-to-good match with PCRctic primers and therefore high probability of successful detection.

Conclusion: PCRctic has been adapted for NHS molecular diagnostics laboratory and is entering a feasibility trial with a tertiary neonatal unit with a multi-centre cross-sectional study to follow. Successful translation of advances in molecular diagnosis into neonatal practice will require concerted efforts of neonatologists, public health specialists, molecular biologists, and biotech

More to explorer

2020 Spring Meeting

20th March 2020 The Royal Society of Medicine, London Open to all professionals working in the field of perinatal medicine welcome. Admission

2019 Summer Meeting

60th anniversary celebration This special meeting marked the 60th anniversary of the founding of the Neonatal Society. A series of keynote lectures

2019 Autumn Meeting

7th November 2019 The Royal Society of Medicine, London 9:30 – 17:30 with a drinks Reception at 18:00 Open to all professionals

Search by category