Abstract
During COVID-19 pandemics, the availability of testing has often been a limiting factor during patient admissions into the hospital. To circumvent this problem, we adapted an existing diagnostic assay, Seegene Allplex SARS-CoV-2, into a point-of-care-style direct qPCR (POC dqPCR) assay and implemented it in the Emergency Department of Clinical Hospital Center Rijeka, Croatia. In a 4-month analysis, we tested over 10,000 patients and demonstrated that POC-dqPCR is robust and reliable and can be successfully implemented in emergency departments and similar near-patient settings and can be performed by medical personnel with little prior experience in qPCR.
Highlights
Accurate and fast diagnosis is a cornerstone of medicine in general and is especially important during pandemics
To investigate whether the Seegene Allplex SARS-CoV-2 assay could be adapted into a dqPCR assay for SARS-CoV-2 detection utilizing the protocol schematically depicted in Figure 1, we compared the performance of Seegene’s recommended assay procedure at the time with the modified, direct Quantitative PCR (qPCR) assay in which oropharyngeal swab samples were directly subjected to qPCR analysis without performing viral RNA isolation and purification
(77) were representative of the populations we expected the emergency departments (EDs) to analyze on a routine basis, such as patients arriving at the ED with COVID-19-like symptoms, patients requiring a SARS-CoV-2 test for hospital admission, or CHC Rijeka personnel working with confirmed or suspected COVID-19 patients
Summary
Accurate and fast diagnosis is a cornerstone of medicine in general and is especially important during pandemics. As the incidence of infection increases, a high burden is placed on the hospitals to quickly and accurately identify infectious patients and personnel to prevent intra-hospital transmission of a disease that could be fatal to sensitive patients and jeopardize the availability and quality of healthcare. Quantitative PCR (qPCR) based diagnostic procedures, and associated primers and probes, were developed within a month of the Wuhan outbreak and have remained the gold standard for SARS-CoV-2 diagnosis due to their high sensitivity, specificity, and suitability for various types of samples [3,4,5]. QPCR-testing for SARS-CoV2 requires highly trained personnel and dedicated BSL-2 facilities suited for RNA isolation and diagnostic qPCR.
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