Multiplex Solid-Phase Real-Time Polymerase Chain Reaction without DNA Extraction
نویسندگان
چکیده
PurposeCurrent polymerase chain reaction (PCR) methods for the diagnosis of infections are time consuming and require large sample volume skilled technicians. We developed a novel, easy-to-use, rapid (processing time, 1 minute; total 33 minutes) multiplex real-time PCR test (Direct Strip PCR) that did not DNA extraction to detect 9 pathogens could cause uveitis in 20-μl samples.DesignMulticenter prospective evaluation diagnostic test.ParticipantsA 511 participants (patients with infectious controls) were examined at 18 institutes worldwide.MethodsAfter validation, intraocular fluid samples subjected etiologic or exclusive diagnosis, including intraoperative diagnosis.Main Outcome MeasuresThe concordance correlations between Direct quantitative (qPCR) results.ResultsDirect exhibited detection, good repeatability specificity, long storage stability, detection ability equal qPCR. It also showed low interinstitutional variability compared qPCR, even when beginners used various machines. The high against qPCR (positive rate, 98.8%–100%; negative 99.8%–100%; κ coefficient, 0.969–1.000; P < 0.001–0.031). Additionally, results obtained using highly correlated (ρ = 0.748; 0.001). This assay was diagnosis.ConclusionsThe may improve prognosis diseases because it facilitates first hospital visit can be diagnosis. Current samples. Multicenter test. A worldwide. After results.
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ژورنال
عنوان ژورنال: Ophthalmology
سال: 2021
ISSN: ['2468-7162', '2468-7170']
DOI: https://doi.org/10.1016/j.ophtha.2020.09.028