@article{mbs:/content/journal/jmm/10.1099/jmm.0.000154, author = "McHugh, Martin P. and Gray, Steve J. and Kaczmarski, Edward B. and Guiver, Malcolm", title = "Reduced turnaround time and improved diagnosis of invasive serogroup B Neisseria meningitidis and Streptococcus pneumoniae infections using a lyophilized quadruplex quantitative PCR", journal= "Journal of Medical Microbiology", year = "2015", volume = "64", number = "11", pages = "1321-1328", doi = "https://doi.org/10.1099/jmm.0.000154", url = "https://www.microbiologyresearch.org/content/journal/jmm/10.1099/jmm.0.000154", publisher = "Microbiology Society", issn = "1473-5644", type = "Journal Article", abstract = "Since 1996 the Meningococcal Reference Unit (MRU) in Manchester has provided a national service for PCR confirmation of meningococcal and pneumococcal disease. Neisseria meningitidis serogroup B is predominant in the UK, accounting for >60 % of cases. In response to this, the MRU has developed a quadruplex quantitative PCR that detects N. meningitidis capsule transporter (ctrA), serogroup B sialyltransferase (siaD), Streptococcus pneumoniae pneumolysin (ply) and an internal control. The assay was prepared in a ready-to-use lyophilized format by Applied Biosystems. Laboratory validation showed excellent performance in a specificity panel of 52 isolates and improved detection in comparison with the routine assay. Testing of 244 patient samples showed sensitivity of 93 % [95 % confidence interval (CI): 88–98 %] for the ctrA assay, 95 % (95 % CI: 91–100 %) for the siaD assay and 100 % (95 % CI: 95–100 %) for the ply assay. Specificity was 100 % (95 % CI: 98–100 %) for both meningococcal targets and 95 % (95 % CI: 92–98 %) for ply. The quadruplex also retained high performance in mixed samples and had acceptable reproducibility. After introduction of the quadruplex into routine use the turnaround time for N. meningitidis group B PCR confirmation reduced from 37 to 29 h and the internal control has proved useful for detecting inhibitory samples. The quadruplex assay provides rapid group B confirmation of meningococcal positive samples, enabling timely public health interventions for the most common disease-causing meningococcal serogroup in the UK.", }