@conference{
author = "Blagojević, Danijela and Milanović, Ivana and Imširović, Mirela and Lazić, Sanda and Maksimović, Zlatko and Tanić, Nasta and Tanić, Nikola",
year = "2024",
abstract = "COVID-19 pandemic caused by coronavirus 2 respiratory virus (SARS-CoV-2) required fast and effective diagnostics for the purpose of preventing the spread of the disease and outbreak monitoring and control. Therefore, prompt, accurate and reliable testing was of particular importance. Indubitably, the most representative, the most sensitive and the fastest method for detection of SARS CoV-2 is Real Time RT-PCR (RT-qPCR). During the pandemic there were numerous different assays, reagents, protocols and reporting procedures under the question mark because of lack of certified standards, standardization of RNA extraction and reporting procedures. In practice, the reliability of RT-qPCR results depend on a number of parameters that include sample collection and processing, method of RNA extraction, choice of assay, choice of instrument, analysis method and operator intervention. Here we present the various challenges which we encountered during our work and emphasize comparative analysis by different assays, as well as automated versus manual RNA extraction. Sampling, adequate manipulation load, the quality of the assay and interpretation of the result are extremely important. Our results revealed that manual viral RNA extraction should be a method of choice for high sensitivity. In addition, amplification assays targeting three SARS-CoV-2 genes are much more efficient from those targeting one. Unfortunately, RT-qPCR (originally quantitative method) was exclusively used as qualitative diagnostic test for SARS-CoV-2. We think that the ideal testing regimen would involve not just qualitative detection of SARS-CoV-2 but reliable and meaningful quantitative reporting of viral load. For such a thing, a consensus at the level of the world medical community is necessary regarding the most important thing: defining the cut-off value for a clinically significant viral load. It has never been done and therefore many people, all over the world, were sentenced to house arrest for no reason at all.",
journal = "Genetics & Applications",
title = "Challenges in SARS-COV-2 diagnostics by real time RT-PCR",
pages = "33-33",
url = "https://hdl.handle.net/21.15107/rcub_vinar_13316"
}