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10.1371/journal.pone.0246647 http://scihub22266oqcxt.onion/10.1371/journal.pone.0246647 33534838!7857565!33534838     free     free     free Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       PLoS+One 2021 ; 16 (2): e0246647 Nephropedia Template TP {{tp|p=33534838|t=2021. Open-source RNA extraction and RT-qPCR methods for SARS-CoV-2 detection |pdf=|usr=}}{{33534838}} gab.com Text Open-source RNA extraction and RT-qPCR methods for SARS-CoV-2 detection JO: PLoS One http://www.kidney.de/pget.php?&tw=1&pmid=33534838 #FOAvip Re-opening of communities in the midst of the ongoing COVID-19 pandemic has ignited new waves of infections in many places around the world. Mitigating the risk of reopening will require widespread SARS-CoV-2 testing, which would be greatly facilitated by simple, rapid, and inexpensive testing methods. This study evaluates several protocols for RNA extraction and RT-qPCR that are simpler and less expensive than prevailing methods. First, isopropanol precipitation is shown to provide an effective means of RNA extraction from nasopharyngeal (NP) swab samples. Second, direct addition of NP swab samples to RT-qPCRs is evaluated without an RNA extraction step. A simple, inexpensive swab collection solution suitable for direct addition is validated using contrived swab samples. Third, an open-source master mix for RT-qPCR is described that permits detection of viral RNA in NP swab samples with a limit of detection of approximately 50 RNA copies per reaction. Quantification cycle (Cq) values for purified RNA from 30 known positive clinical samples showed a strong correlation (r2 = 0.98) between this homemade master mix and commercial TaqPath master mix. Lastly, end-point fluorescence imaging is found to provide an accurate diagnostic readout without requiring a qPCR thermocycler. Adoption of these simple, open-source methods has the potential to reduce the time and expense of COVID-19 testing. Twit Text FOAVip Open-source RNA extraction and RT-qPCR methods for SARS-CoV-2 detection ????PLoS One http://www.kidney.de/pget.php?&tw=1&pmid=33534838 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=33534838 #FOAvip English Wikipedia <ref>{{Cite pmid|33534838}}</ref> Open-source RNA extraction and RT-qPCR methods for SARS-CoV-2 detection #MMPMID33534838 Graham TGW; Dugast-Darzacq C; Dailey GM; Nguyenla XH; Van Dis E; Esbin MN; Abidi A; Stanley SA; Darzacq X; Tjian R PLoS One 2021[]; 16 (2): e0246647 PMID33534838show ga Re-opening of communities in the midst of the ongoing COVID-19 pandemic has ignited new waves of infections in many places around the world. Mitigating the risk of reopening will require widespread SARS-CoV-2 testing, which would be greatly facilitated by simple, rapid, and inexpensive testing methods. This study evaluates several protocols for RNA extraction and RT-qPCR that are simpler and less expensive than prevailing methods. First, isopropanol precipitation is shown to provide an effective means of RNA extraction from nasopharyngeal (NP) swab samples. Second, direct addition of NP swab samples to RT-qPCRs is evaluated without an RNA extraction step. A simple, inexpensive swab collection solution suitable for direct addition is validated using contrived swab samples. Third, an open-source master mix for RT-qPCR is described that permits detection of viral RNA in NP swab samples with a limit of detection of approximately 50 RNA copies per reaction. Quantification cycle (Cq) values for purified RNA from 30 known positive clinical samples showed a strong correlation (r2 = 0.98) between this homemade master mix and commercial TaqPath master mix. Lastly, end-point fluorescence imaging is found to provide an accurate diagnostic readout without requiring a qPCR thermocycler. Adoption of these simple, open-source methods has the potential to reduce the time and expense of COVID-19 testing. |COVID-19 Nucleic Acid Testing[MESH] |COVID-19/*diagnosis/virology[MESH] |Chemical Precipitation[MESH] |Coronavirus Nucleocapsid Proteins/genetics[MESH] |Humans[MESH] |Limit of Detection[MESH] |Nasopharynx/virology[MESH] |Phosphoproteins/genetics[MESH] |RNA, Viral/*analysis/isolation & purification/metabolism[MESH] |Real-Time Polymerase Chain Reaction/*methods[MESH]Open-source RNA extraction and RT-qPCR methods for SARS-CoV-2 detectio(epmc).pdf DeepDyve Pubget Overpricing