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10.1016/j.ebiom.2021.103236 http://scihub22266oqcxt.onion/10.1016/j.ebiom.2021.103236 33582488!7878117!33582488     free     free     free Deprecated: Implicit conversion from float 229.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 229.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 229.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 229.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       EBioMedicine 2021 ; 64 (ä): 103236 Nephropedia Template TP {{tp|p=33582488|t=2021. Simpler and faster Covid-19 testing: Strategies to streamline SARS-CoV-2 molecular assays |pdf=|usr=}}{{33582488}} gab.com Text Simpler and faster Covid-19 testing: Strategies to streamline SARS-CoV-2 molecular assays JO: EBioMedicine http://www.kidney.de/pget.php?&tw=1&pmid=33582488 #FOAvip BACKGROUND: Detection of SARS-CoV-2 infections is important for treatment, isolation of infected and exposed individuals, and contact tracing. RT-qPCR is the "gold-standard" method to sensitively detect SARS-CoV-2 RNA, but most laboratory-developed RT-qPCR assays involve complex steps. Here, we aimed to simplify RT-qPCR assays by streamlining reaction setup, eliminating RNA extraction, and proposing reduced-cost detection workflows that avoid the need for expensive qPCR instruments. METHOD: A low-cost RT-PCR based "kit" was developed for faster turnaround than the CDC developed protocol. We demonstrated three detection workflows: two that can be deployed in laboratories conducting assays of variable complexity, and one that could be simple enough for point-of-care. Analytical sensitivity was assessed using SARS-CoV-2 RNA spiked in simulated nasal matrix. Clinical performance was evaluated using contrived human nasal matrix (n = 41) and clinical nasal specimens collected from individuals with respiratory symptoms (n = 110). FINDING: The analytical sensitivity of the lyophilised RT-PCR was 10 copies/reaction using purified SARS-CoV-2 RNA, and 20 copies/reaction when using direct lysate in simulated nasal matrix. Evaluation of assay performance on contrived human matrix showed 96.7-100% specificity and 100% sensitivity at >/=20 RNA copies. A head-to-head comparison with the standard CDC protocol on clinical specimens showed 83.8-94.6% sensitivity and 96.8-100% specificity. We found 3.6% indeterminate samples (undetected human control), lower than 8.1% with the standard protocol. INTERPRETATION: This preliminary work should support laboratories or commercial entities to develop and expand access to Covid-19 testing. Software guidance development for this assay is ongoing to enable implementation in other settings. FUND: USA NIH R01AI140845 and Seattle Children's Research Institute. Twit Text FOAVip Simpler and faster Covid-19 testing: Strategies to streamline SARS-CoV-2 molecular assays ????EBioMedicine http://www.kidney.de/pget.php?&tw=1&pmid=33582488 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=33582488 #FOAvip English Wikipedia <ref>{{Cite pmid|33582488}}</ref> Simpler and faster Covid-19 testing: Strategies to streamline SARS-CoV-2 molecular assays #MMPMID33582488 Panpradist N; Wang Q; Ruth PS; Kotnik JH; Oreskovic AK; Miller A; Stewart SWA; Vrana J; Han PD; Beck IA; Starita LM; Frenkel LM; Lutz BR EBioMedicine 2021[Feb]; 64 (ä): 103236 PMID33582488show ga BACKGROUND: Detection of SARS-CoV-2 infections is important for treatment, isolation of infected and exposed individuals, and contact tracing. RT-qPCR is the "gold-standard" method to sensitively detect SARS-CoV-2 RNA, but most laboratory-developed RT-qPCR assays involve complex steps. Here, we aimed to simplify RT-qPCR assays by streamlining reaction setup, eliminating RNA extraction, and proposing reduced-cost detection workflows that avoid the need for expensive qPCR instruments. METHOD: A low-cost RT-PCR based "kit" was developed for faster turnaround than the CDC developed protocol. We demonstrated three detection workflows: two that can be deployed in laboratories conducting assays of variable complexity, and one that could be simple enough for point-of-care. Analytical sensitivity was assessed using SARS-CoV-2 RNA spiked in simulated nasal matrix. Clinical performance was evaluated using contrived human nasal matrix (n = 41) and clinical nasal specimens collected from individuals with respiratory symptoms (n = 110). FINDING: The analytical sensitivity of the lyophilised RT-PCR was 10 copies/reaction using purified SARS-CoV-2 RNA, and 20 copies/reaction when using direct lysate in simulated nasal matrix. Evaluation of assay performance on contrived human matrix showed 96.7-100% specificity and 100% sensitivity at >/=20 RNA copies. A head-to-head comparison with the standard CDC protocol on clinical specimens showed 83.8-94.6% sensitivity and 96.8-100% specificity. We found 3.6% indeterminate samples (undetected human control), lower than 8.1% with the standard protocol. INTERPRETATION: This preliminary work should support laboratories or commercial entities to develop and expand access to Covid-19 testing. Software guidance development for this assay is ongoing to enable implementation in other settings. FUND: USA NIH R01AI140845 and Seattle Children's Research Institute. |*COVID-19 Nucleic Acid Testing[MESH] |*COVID-19/diagnosis/genetics[MESH] |*Real-Time Polymerase Chain Reaction[MESH] |*Reverse Transcriptase Polymerase Chain Reaction[MESH] |Humans[MESH] |RNA, Viral/*genetics[MESH] |SARS-CoV-2/*genetics[MESH]Simpler and faster Covid-19 testing: Strategies to streamline SARS-CoV(epmc).pdf DeepDyve Pubget Overpricing