Nucleic acid amplification tests are the model for detecting SARS-CoV-2, but efficient extraction of viral RNA is essential. Here, David Mathias and Lucy Goulart compare the performance and usability of two RNA extraction kits.
Coronavirus disease 2019 (COVID-19) is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and was first reported in Wuhan, China. COVID-19 is a highly infectious disease and rapidly spread to other countries throughout the world.
In order to minimise the spread of SARS-CoV-2, early and accurate detection has been critical. Reverse transcriptase real-time PCR (rRT-PCR), a type of nucleic acid amplification test (NAAT), is still considered the gold standard in detecting SARS-CoV-2 infection. Compared to other available viral detection methods, rRT-PCR is highly sensitive and specific.2 As well as being faster, using rRT-PCR as opposed to viral antigen detection, serology or electron microscopy allows for a lower potential for errors and/or contamination. Many rRT-PCR testing kits have been produced around the world and have become commercially available. With the initial need for high volumes of testing becoming less, laboratories are now considering ways in which to effectively augment their COVID-19 detection workflow for the greatest performance.
To carry out rRT-PCR for detection of SARS-CoV-2, RNA first has to be isolated from a sample using one of a number of different extraction methods available. Nucleic acid (NA) extraction in its most basic form includes three main steps: release or extraction of the NA from a sample by lysis, isolation or separation of the NA from unwanted sample material, and removal of inhibitory substances to purify the NA. The efficiency of the viral RNA extraction procedure used can greatly affect rRT-PCR assay performance in detecting positive samples.3
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