Meningitis and encephalitis are inflammatory, often infectious, processes of the central nervous system that can result in significant morbidity and mortality. Prompt, appropriate therapy is crucial, but determining the infectious etiology can be difficult and time-consuming. A wide-range of pathogens can be involved including both bacteria and viruses. Culture is the standard for diagnosis of bacterial Meningoencephalitis (ME), PCR of viral nucleic acid is the standard for "aseptic" causes. However the current methods are either 1) too slow to be clinically relevant; 2) technically complex; 3) limited in the number of targets r 4) not cleared by the FDA. Idaho Technology (ITI) has developed a "lab in a pouch" PCR-based diagnostic system termed "FilmArray" that is rapid, easy to use, and can test for large panels of infectious agents simultaneously. A FilmArray pouch that can detect 15 respiratory pathogens was cleared by the FDA as a CLIA "moderate complexity" test. We propose to apply the FilmArray technology to the problem of ME diagnosis. SA1: Develop a FilmArray Meningoencephalitis (FAME) panel: We will construct a FilmArray pouch that combines PCR assays from existing panels with five new assays to detect the following ME-causing organisms directly from cerebrospinal fluid (CSF): Enterobacteriaceae, E. coli, Enterococcus species, H. influenza, L. monocytogenes, Mycoplasma pneumoniae, N. meningitidis, P. aeruginosa, Staphylococci species, S. aureus, Streptococcus species, S. agalactiae, S. pneumoniae, S. pyogenes, viridians streptococci, Adenovirus, Enterovirus, Parechovirus, HSV1, HSV2, VZV, EBV, CMV, HHV-6. SA2: Optimize nucleic acid extraction for virus and bacteria from CSF: CSF is one of the less complex sample matrices that have been processed on the FilmArray (when compared to nasal swabs, blood culture and stool). However, detecting bacteria, and virus, directly from CSF, may require the highest possible sensitivity. We will optimize the nucleic acid purification steps internal to the pouch as well as investigate simple steps to concentrate bacteria and viruses before the pouch SA3: Test the FAME panel on 300 CSF samples: Our collaborators will test CSF samples from pediatric and adult patients with suspected ME. We will compare these results to their standard assays performed on the samples, supplemented with the results of the comparator assays described in SA1. A successful outcome of this proposal will be a FilmArray pouch capable of detecting and identifying the common ME pathogens. It will be ready for the analytical and clinical trials necessary for a 510(k) submission to the FDA. This would be the subject of a future phase II submission.
Public Health Relevance Statement: Public Health Relevance: Meningitis and encephalitis are life threatening inflammations of the brain and spinal cord which are often caused by infectious agents. Rapid, comprehensive, diagnosis of the virus or bacteria responsible for these conditions is critical for determining the most effective treatment. We will apply Idaho Technology's automated, multiplex nucleic acid test platform (the FilmArray) to the problem of meningoencephalitis diagnosis.
Project Terms: Acute; Adenoviruses; Adult; Affect; Bacteria; base; Biological; Biological Assay; Blood; Buffers; Caring; Cerebrospinal Fluid; Childhood; Clinical; clinical practice; Clinical Sensitivity; Clinical Trials; clinically relevant; CNS processing; Complex; Culture Media; Cytomegalovirus; Detection; Development; Diagnosis; diagnosis standard; Diagnostic; DNA-Directed DNA Polymerase; effective therapy; Encephalitis; Enterobacteriaceae; Enterococcus; Enterovirus; Escherichia coli; Etiology; Evaluation; Feces; Future; Genus staphylococcus; Gold; Haemophilus influenzae; Herpesvirus 1, Human; Hour; Housing; Human Herpesvirus 2; Human Herpesvirus 4; Human Herpesvirus 6; Idaho; In Vitro; Infection; Infectious Agent; Inflammatory; Laboratories; Life; Listeria monocytogenes; Medical; Meningitis; Meningoencephalitis; Methods; Morbidity - disease rate; Mortality Vital Statistics; Mycoplasma pneumoniae; Neisseria meningitidis; new technology; Nose; Nucleic Acid Amplification Tests; nucleic acid purification; Nucleic Acids; Organism; Outcome; Parechovirus; pathogen; Pathogen detection; Patients; Performance; Phase; Polymerase Chain Reaction; Population; preclinical evaluation; Process; Protocols documentation; Pseudomonas aeruginosa; public health relevance; Publishing; rapid detection; Resources; respiratory; Sampling; Sensitivity and Specificity; skills; Soil; Spinal Cord; Staphylococcus aureus; Streptococcus; Streptococcus Group B; Streptococcus pneumoniae; Streptococcus pyogenes; Swab; System; Technology; Testing; Time; United States Food and Drug Administration; Viral; viral detection; viral DNA; Virus
