Automating laboratory developed tests using the panther fusion open access: Analytical and clinical evaluation of assays for the detection of viral causes of meningitis and encephalitis.

内容摘要

Laboratory developed tests (LDTs) are high-complexity tests developed by individual laboratories. LDTs have been critical for addressing clinical needs not available commercially. Automation facilitates workflow efficiency and expansion of testing menus. At the Johns Hopkins Medical Microbiology Laboratory, different automation solutions were evaluated for molecular LDTs to transition manual real-time PCR diagnostics to automated platforms. This study evaluated the performance of the Panther Fusion Open Access platform, with a focus on assays developed for viral infections of the central nervous system (CNS). Molecular assays for HSV-1, HSV-2, VZV, enterovirus, and parechovirus were developed on the Panther Fusion platform using Open Access. Analytical studies included determination of sensitivity, specificity, inclusivity, onboard reagent stability, and agreement. Clinical evaluations were performed and compared to standard-of-care assays, including the BioFire Meningitis/Encephalitis panel and DiaSorin Simplexa HSV-1&2 and VZV Direct assays. Analytical sensitivities were comparable or better than standard-of-care tests. A blind panel of 297 unique samples demonstrated 100% agreement and reproducibility. Among 101 clinical samples tested retrospectively and prospectively, the developed viral assays showed 100% positive and 98.8% negative agreement with comparator assays. Patients with positive viral results were more frequently treated with antivirals rather than antibiotics, had final diagnoses of viral meningitis or encephalitis, and exhibited significantly higher cerebrospinal fluid (CSF) mononuclear cell counts, supporting the clinical validity of developed and comparator assays. Assays developed on the Panther Fusion Open Access platform demonstrate excellent analytical and clinical performance for CNS viral targets. The platform provides a highly efficient, fully automated open-access solution for LDTs in high-complexity laboratories.