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10.1093/clinchem/hvab132 http://scihub22266oqcxt.onion/10.1093/clinchem/hvab132 34286830!8406859!34286830     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       Clin+Chem 2021 ; 68 (1): 143-152 Nephropedia Template TP {{tp|p=34286830|t=2021. SwabExpress: An End-to-End Protocol for Extraction-Free COVID-19 Testing |pdf=|usr=}}{{34286830}} gab.com Text SwabExpress: An End-to-End Protocol for Extraction-Free COVID-19 Testing JO: Clin Chem http://www.kidney.de/pget.php?&tw=1&pmid=34286830 #FOAvip BACKGROUND: The urgent need for massively scaled clinical testing for SARS-CoV-2, along with global shortages of critical reagents and supplies, has necessitated development of streamlined laboratory testing protocols. Conventional nucleic acid testing for SARS-CoV-2 involves collection of a clinical specimen with a nasopharyngeal swab in transport medium, nucleic acid extraction, and quantitative reverse-transcription PCR (RT-qPCR). As testing has scaled across the world, the global supply chain has buckled, rendering testing reagents and materials scarce. To address shortages, we developed SwabExpress, an end-to-end protocol developed to employ mass produced anterior nares swabs and bypass the requirement for transport media and nucleic acid extraction. METHODS: We evaluated anterior nares swabs, transported dry and eluted in low-TE buffer as a direct-to-RT-qPCR alternative to extraction-dependent viral transport media. We validated our protocol of using heat treatment for viral inactivation and added a proteinase K digestion step to reduce amplification interference. We tested this protocol across archived and prospectively collected swab specimens to fine-tune test performance. RESULTS: After optimization, SwabExpress has a low limit of detection at 2-4 molecules/microL, 100% sensitivity, and 99.4% specificity when compared side by side with a traditional RT-qPCR protocol employing extraction. On real-world specimens, SwabExpress outperforms an automated extraction system while simultaneously reducing cost and hands-on time. CONCLUSION: SwabExpress is a simplified workflow that facilitates scaled testing for COVID-19 without sacrificing test performance. It may serve as a template for the simplification of PCR-based clinical laboratory tests, particularly in times of critical shortages during pandemics. Twit Text FOAVip SwabExpress: An End-to-End Protocol for Extraction-Free COVID-19 Testing ????Clin Chem http://www.kidney.de/pget.php?&tw=1&pmid=34286830 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=34286830 #FOAvip English Wikipedia <ref>{{Cite pmid|34286830}}</ref> SwabExpress: An End-to-End Protocol for Extraction-Free COVID-19 Testing #MMPMID34286830 Srivatsan S; Heidl S; Pfau B; Martin BK; Han PD; Zhong W; van Raay K; McDermot E; Opsahl J; Gamboa L; Smith N; Truong M; Cho S; Barrow KA; Rich LM; Stone J; Wolf CR; McCulloch DJ; Kim AE; Brandstetter E; Sohlberg SL; Ilcisin M; Geyer RE; Chen W; Gehring J; Kosuri S; Bedford T; Rieder MJ; Nickerson DA; Chu HY; Konnick EQ; Debley JS; Shendure J; Lockwood CM; Starita LM Clin Chem 2021[Dec]; 68 (1): 143-152 PMID34286830show ga BACKGROUND: The urgent need for massively scaled clinical testing for SARS-CoV-2, along with global shortages of critical reagents and supplies, has necessitated development of streamlined laboratory testing protocols. Conventional nucleic acid testing for SARS-CoV-2 involves collection of a clinical specimen with a nasopharyngeal swab in transport medium, nucleic acid extraction, and quantitative reverse-transcription PCR (RT-qPCR). As testing has scaled across the world, the global supply chain has buckled, rendering testing reagents and materials scarce. To address shortages, we developed SwabExpress, an end-to-end protocol developed to employ mass produced anterior nares swabs and bypass the requirement for transport media and nucleic acid extraction. METHODS: We evaluated anterior nares swabs, transported dry and eluted in low-TE buffer as a direct-to-RT-qPCR alternative to extraction-dependent viral transport media. We validated our protocol of using heat treatment for viral inactivation and added a proteinase K digestion step to reduce amplification interference. We tested this protocol across archived and prospectively collected swab specimens to fine-tune test performance. RESULTS: After optimization, SwabExpress has a low limit of detection at 2-4 molecules/microL, 100% sensitivity, and 99.4% specificity when compared side by side with a traditional RT-qPCR protocol employing extraction. On real-world specimens, SwabExpress outperforms an automated extraction system while simultaneously reducing cost and hands-on time. CONCLUSION: SwabExpress is a simplified workflow that facilitates scaled testing for COVID-19 without sacrificing test performance. It may serve as a template for the simplification of PCR-based clinical laboratory tests, particularly in times of critical shortages during pandemics. |*COVID-19/diagnosis[MESH] |COVID-19 Nucleic Acid Testing/*methods[MESH] |Clinical Laboratory Techniques[MESH] |Humans[MESH] |RNA, Viral/isolation & purification[MESH] |Real-Time Polymerase Chain Reaction[MESH] |SARS-CoV-2/isolation & purification[MESH] |Sensitivity and Specificity[MESH]SwabExpress: An End-to-End Protocol for Extraction-Free COVID-19 Testi(epmc).pdf DeepDyve Pubget Overpricing