Meningitis is associated with high morbidity and mortality if not diagnosed in a timely manner. Diagnosis of meningitis by culture is time consuming, may lack sensitivity, requires high-level technical expertise; and in the case of bacterial infection, may return false-negative results if antimicrobial therapy has been initiated. Early identification of causative bacterial and viral agents could aid in the prompt and proper treatment of meningitis, potentially preventing adverse clinical outcomes. To this end, we are developing panels of multiplexed qPCR assays using our novel detection technology to assist with the rapid detection of 10 bacterial and viral agents associated with meningitis.

Multiplexed qPCR tests have been developed using MNAzyme detection technology. In the presence of target, MNAzymes assemble from component DNA oligonucleotides. These cleave universal probes thus generating signal in real-time. MNAzyme qPCR possesses superior specificity and multiplex capacity compared to commonly used real-time chemistries.  The test includes four unique panels which detect the following organisms: (1) Neisseria meningitidis, Streptococcus pneumoniae and Hemophilus  influenzae, (2) Group B streptococcus, Escherichia coli and Listeria monocytogenes, (3) Herpes simplex virus type 1, Herpes simplex virus type 2 and Varicella zoster virus, and (4) Enterovirus. The multiplex panels were designed for compatibility across commonly used “open” qPCR platforms and all have been evaluated for dynamic range, analytical specificity and sensitivity and cross reactivity.

The panels demonstrated robust analytical specificity for the target organisms, with no cross reactivity with other assay or panel targets, closely related organisms or organisms that generate similar symptoms. All target assays had a limit of detection of at least 50 copies per reaction as determined by Probit analysis.

MNAzyme qPCR provides a flexible and unique approach to qPCR that is specific, sensitive, rapid and easily multiplexed. The assays developed are useful for rapid identification of important meningitis pathogens.