Abstract
We present the RespiDisk enabling the fully automated and multiplex point-of-care (POC) detection of (currently) up to 19 respiratory tract infection (RTI) pathogens from a single sample based on reverse transcriptase polymerase chain reaction (RT-PCR). RespiDisk comprises a RTI-specific implementation of the centrifugal microfluidic LabDisk platform and combines new and existing advanced unit operations for liquid control, thereby automating all assay steps only by a spinning frequency and temperature protocol in combination with the use of a permanent magnet for in situ bead handing. The capabilities of the system were demonstrated with 36 tested quality samples mimicking clinical conditions (clinical and/or cultured material suspended in transport medium or synthetic bronchoalveolar lavage (BAL)) from past external quality assessment (EQA) panels covering 13 of the 19 integrated RTI detection assays. In total, 36 samples × 19 assays/sample resulting in 684 assays were performed with the RespiDisk, and its analytical performance was in full agreement with the routine clinical workflow serving as reference. A strong feature of the platform is its universality since its components allow the simultaneous detection of a broad panel of bacteria and viruses in a single run, thereby enabling the differentiation between antibiotic-treatable diseases. Furthermore, the full integration of all necessary biochemical components enables a reduction of the hands-on time from manual to automated sample-to-answer analysis to about 5 min. The study was performed on an air-heated LabDisk Player instrument with a time-to-result of 200 min.
Highlights
Lower respiratory tract infections (RTIs) are a major cause of hospitalizations among adults and children globally, resultingIn several occasions, such as in Emergency Rooms or in remote clinics, the diagnosis of RTI pathogens can be especially challenging[4] as it must be done rapidly and at the patient side instead of following the routine procedure of sample collection and dispatching it to a central laboratory for analysis
For the multiple pathogen programmes (i.e. RESPI18S and RESPII17S), only datasets from laboratories that included the specific pathogens in their molecular workflows were used to calculate the percentage of correct results reported within these external quality assessment (EQA) programmes (# returned EQA datasets from participants: RESPI18S n = 171; RESPII17S n = 126; LPDNA18S n = 153; MP18S n = 204)
The proof-of-principle study successfully demonstrated the capacity of the developed RespiDisk to provide differentiation between viral and bacterial pathogens causing RTIs in a single test run
Summary
Lower respiratory tract infections (RTIs) are a major cause of hospitalizations among adults and children globally, resulting. In several occasions, such as in Emergency Rooms or in remote clinics, the diagnosis of RTI pathogens can be especially challenging[4] as it must be done rapidly and at the patient side instead of following the routine procedure of sample collection and dispatching it to a central laboratory for analysis. In several cases, empirical or merely symptomatic diagnosis is still used due to the lack of suitable tools and/or the pressure of time (e.g. circumventing culture-based methods due to time constraints).[10] indicators supporting the symptomatic diagnosis such as the medical history, the duration of symptoms, the presence of fever or the auscultation of the lung, lack sufficient specificity to distinguish between a viral or bacterial infection
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