Publication

Development of a rapid recombinase polymerase amplification assay for the detection of Streptococcus pneumoniae in whole blood

Clancy, Eoin
; 
Higgins, Owen
; 
Boo, Teck Wee
; 
Cormican, Martin
; 
Barry, Thomas
; 
Smith, Terry J.
Thumbnail Image
Files
Thumbnail Image
2015_Clancy_E_et_al_BMC_Inf_Dis.pdf
Adobe PDF, 1.12 MB
Repository DOI
Publication Date
2015-10-29
Keywords
Streptococcus pneumoniae, Recombinase Polymerase Amplification, Leader peptidase A, Molecular diagnostics, Real-time PCR, Viridans group streptococci, Pneumococcal pneumonia, Elongation factor, Chain reaction, LYTA gene, Diagnosis, DNA, Samples, Disease
Type
Article
Downloads
Show all metadata
Citation
Clancy, E., Higgins, O., Forrest, M. S., Boo, T. W., Cormican, M., Barry, T., . . . Smith, T. J. (2015). Development of a rapid recombinase polymerase amplification assay for the detection of Streptococcus pneumoniae in whole blood. BMC Infectious Diseases, 15(1), 1-11.
Abstract
Background: Streptococcus pneumoniae is an important cause of microbial disease in humans. The introduction of multivalent vaccines has coincided with a dramatic decrease in the number of pneumococcal-related deaths. In spite of this, at a global level, pneumococcal infection remains an important cause of death among children under 5 years of age and in adults 65 years of age or older. In order to properly manage patients and control the spread of infection, a rapid and highly sensitive diagnostic method is needed for routine implementation, especially in resource-limited regions where pneumococcal disease is most prevalent.Methods: Using the gene encoding leader peptidase A as a molecular diagnostics target, a real-time RPA assay was designed and optimised for the detection of S. pneumoniae in whole blood. The performance of the assay was compared to real-time PCR in terms of its analytical limit of detection and specificity. The inhibitory effect of human genomic DNA on amplification was investigated. The potential clinical utility of the assay was investigated using a small number of clinical samples.Results: The RPA assay has a limit of detection equivalent to PCR (4.0 and 5.1 genome equivalents per reaction, respectively) and was capable of detecting the equivalent of
Funder
|~|
Publisher
BioMed Central
Publisher DOI
10.1186/s12879-015-1212-5
Rights
Attribution-NonCommercial-NoDerivs 3.0 Ireland
cbn
Collections
National Centre for Biomedical Engineering Science (Scholarly Articles)