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10.1371/journal.pone.0243266 http://scihub22266oqcxt.onion/10.1371/journal.pone.0243266 33284857!7721139!33284857     free     free     free Deprecated: Implicit conversion from float 209.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 209.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       PLoS+One 2020 ; 15 (12): e0243266 Nephropedia Template TP {{tp|p=33284857|t=2020. Molecular detection of SARS-CoV-2 using a reagent-free approach |pdf=|usr=}}{{33284857}} gab.com Text Molecular detection of SARS-CoV-2 using a reagent-free approach JO: PLoS One http://www.kidney.de/pget.php?&tw=1&pmid=33284857 #FOAvip Shortage of reagents and consumables required for the extraction and molecular detection of SARS-CoV-2 RNA in respiratory samples has led many laboratories to investigate alternative approaches for sample preparation. Many groups recently presented results using heat processing method of respiratory samples prior to RT-qPCR as an economical method enabling an extremely fast streamlining of the processes at virtually no cost. Here, we present our results using this method and highlight some major pitfalls that diagnostics laboratories should be aware of before proceeding with this methodology. We first investigated various treatments using different temperatures, incubation times and sample volumes to optimise the heat treatment conditions. Although the initial data confirmed results published elsewhere, further investigations revealed unexpected inhibitory properties of some commonly used universal transport media (UTMs) on some commercially available RT-qPCR mixes, leading to a risk of reporting false-negative results. This emphasises the critical importance of a thorough validation process to determine the most suitable reagents to use depending on the sample types to be tested. In conclusion, a heat processing method is effective with very consistent Ct values and a sensitivity of 96.2% when compared to a conventional RNA extraction method. It is also critical to include an internal control to check each sample for potential inhibition. Twit Text FOAVip Molecular detection of SARS-CoV-2 using a reagent-free approach ????PLoS One http://www.kidney.de/pget.php?&tw=1&pmid=33284857 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=33284857 #FOAvip English Wikipedia <ref>{{Cite pmid|33284857}}</ref> Molecular detection of SARS-CoV-2 using a reagent-free approach #MMPMID33284857 Calvez R; Taylor A; Calvo-Bado L; Fraser D; Fink CG PLoS One 2020[]; 15 (12): e0243266 PMID33284857show ga Shortage of reagents and consumables required for the extraction and molecular detection of SARS-CoV-2 RNA in respiratory samples has led many laboratories to investigate alternative approaches for sample preparation. Many groups recently presented results using heat processing method of respiratory samples prior to RT-qPCR as an economical method enabling an extremely fast streamlining of the processes at virtually no cost. Here, we present our results using this method and highlight some major pitfalls that diagnostics laboratories should be aware of before proceeding with this methodology. We first investigated various treatments using different temperatures, incubation times and sample volumes to optimise the heat treatment conditions. Although the initial data confirmed results published elsewhere, further investigations revealed unexpected inhibitory properties of some commonly used universal transport media (UTMs) on some commercially available RT-qPCR mixes, leading to a risk of reporting false-negative results. This emphasises the critical importance of a thorough validation process to determine the most suitable reagents to use depending on the sample types to be tested. In conclusion, a heat processing method is effective with very consistent Ct values and a sensitivity of 96.2% when compared to a conventional RNA extraction method. It is also critical to include an internal control to check each sample for potential inhibition. |COVID-19 Testing/*methods[MESH] |COVID-19/genetics/metabolism[MESH] |Clinical Laboratory Techniques/methods[MESH] |Coronavirus Infections/epidemiology[MESH] |Humans[MESH] |Indicators and Reagents[MESH] |Pandemics[MESH] |Pneumonia, Viral/epidemiology[MESH] |RNA, Viral/genetics[MESH] |Real-Time Polymerase Chain Reaction/methods[MESH] |SARS-CoV-2/genetics/*metabolism[MESH] |Sensitivity and Specificity[MESH] |Specimen Handling/*methods[MESH]Molecular detection of SARS-CoV-2 using a reagent-free approach (epmc).pdf DeepDyve Pubget Overpricing