Abstract
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has caused worldwide losses from all points of view. Although there are efficient vaccines against this virus, rapid detection remains essential for controlling its spread. The gold standard for the COVID-19 diagnostic is the use of quantitative real time polymerase chain reaction (RT-qPCR) from naso-/oro-pharyngeal swabs and, alternatively, saliva specimens. A faster alternative to the RT-qPCR method, the colorimetric reverse transcription loop-mediated isothermal amplification (cLAMP) has a good sensitivity and specificity and has been proposed as an efficient screening method. In this study, we analyzed several sets of LAMP primers for COVID-19 testing using purified RNA samples and raw saliva samples, and tested several reaction conditions. Our results showed that the sensitivity of cLAMP reactions on raw saliva samples can be significantly augmented by the addition of guanidine hydrochloride to levels, which allow its use for COVID-19 screening.
Highlights
With over million infections and over two million fatalities, the SARS-CoV- pandemic heavily impacted the year in basically all socio-economic sectors [, ]
Since in our previous RT-qPCR experiment, only RdRp was correlated to the cLAMP results, we focused on RdRp ampli cation as a control method for cLAMP
The LAMP assay has multiple advantages compared to the RT-PCR method: it eliminates the need for a thermocycler, and the target ampli cation can be monitored by using pH sensitive dyes such as phenol red
Summary
With over million infections and over two million fatalities, the SARS-CoV- pandemic heavily impacted the year in basically all socio-economic sectors [ , ]. In December , both the US Food and Drug Administration (FDA) and the European Medicines Agency (EMA) approved the P zer-BionTech mRNA vaccine for emergency use, and a consistent economic recovery has been witnessed ever since. This was swiftly followed by emergency authorizations of several other vaccines; given the continuous appearance of novel SARS-CoV- mutated strains, the management of the SARS-CoV- pandemic requires the prompt, Timisoara Med. The gold standard for SARS-CoV- diagnosis/detection is quantitative real time polymerase chain reaction (RT-qPCR). The RT-qPCR method is not suitable for large population screening applications [ , ]
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