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10.1038/s41551-020-00603-x http://scihub22266oqcxt.onion/10.1038/s41551-020-00603-x 32848209!ä!32848209 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       Nat+Biomed+Eng 2020 ; 4 (12): 1140-1149 Nephropedia Template TP {{tp|p=32848209|t=2020. Clinical validation of a Cas13-based assay for the detection of SARS-CoV-2 RNA |pdf=|usr=}}{{32848209}} gab.com Text Clinical validation of a Cas13-based assay for the detection of SARS-CoV-2 RNA JO: Nat Biomed Eng http://www.kidney.de/pget.php?&tw=1&pmid=32848209 #FOAvip Nucleic acid detection by isothermal amplification and the collateral cleavage of reporter molecules by CRISPR-associated enzymes is a promising alternative to quantitative PCR. Here, we report the clinical validation of the specific high-sensitivity enzymatic reporter unlocking (SHERLOCK) assay using the enzyme Cas13a from Leptotrichia wadei for the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-the virus that causes coronavirus disease 2019 (COVID-19)-in 154 nasopharyngeal and throat swab samples collected at Siriraj Hospital, Thailand. Within a detection limit of 42 RNA copies per reaction, SHERLOCK was 100% specific and 100% sensitive with a fluorescence readout, and 100% specific and 97% sensitive with a lateral-flow readout. For the full range of viral load in the clinical samples, the fluorescence readout was 100% specific and 96% sensitive. For 380 SARS-CoV-2-negative pre-operative samples from patients undergoing surgery, SHERLOCK was in 100% agreement with quantitative PCR with reverse transcription. The assay, which we show is amenable to multiplexed detection in a single lateral-flow strip incorporating an internal control for ribonuclease contamination, should facilitate SARS-CoV-2 detection in settings with limited resources. Twit Text FOAVip Clinical validation of a Cas13-based assay for the detection of SARS-CoV-2 RNA ????Nat Biomed Eng http://www.kidney.de/pget.php?&tw=1&pmid=32848209 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=32848209 #FOAvip English Wikipedia <ref>{{Cite pmid|32848209}}</ref> Clinical validation of a Cas13-based assay for the detection of SARS-CoV-2 RNA #MMPMID32848209 Patchsung M; Jantarug K; Pattama A; Aphicho K; Suraritdechachai S; Meesawat P; Sappakhaw K; Leelahakorn N; Ruenkam T; Wongsatit T; Athipanyasilp N; Eiamthong B; Lakkanasirorat B; Phoodokmai T; Niljianskul N; Pakotiprapha D; Chanarat S; Homchan A; Tinikul R; Kamutira P; Phiwkaow K; Soithongcharoen S; Kantiwiriyawanitch C; Pongsupasa V; Trisrivirat D; Jaroensuk J; Wongnate T; Maenpuen S; Chaiyen P; Kamnerdnakta S; Swangsri J; Chuthapisith S; Sirivatanauksorn Y; Chaimayo C; Sutthent R; Kantakamalakul W; Joung J; Ladha A; Jin X; Gootenberg JS; Abudayyeh OO; Zhang F; Horthongkham N; Uttamapinant C Nat Biomed Eng 2020[Dec]; 4 (12): 1140-1149 PMID32848209show ga Nucleic acid detection by isothermal amplification and the collateral cleavage of reporter molecules by CRISPR-associated enzymes is a promising alternative to quantitative PCR. Here, we report the clinical validation of the specific high-sensitivity enzymatic reporter unlocking (SHERLOCK) assay using the enzyme Cas13a from Leptotrichia wadei for the detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2)-the virus that causes coronavirus disease 2019 (COVID-19)-in 154 nasopharyngeal and throat swab samples collected at Siriraj Hospital, Thailand. Within a detection limit of 42 RNA copies per reaction, SHERLOCK was 100% specific and 100% sensitive with a fluorescence readout, and 100% specific and 97% sensitive with a lateral-flow readout. For the full range of viral load in the clinical samples, the fluorescence readout was 100% specific and 96% sensitive. For 380 SARS-CoV-2-negative pre-operative samples from patients undergoing surgery, SHERLOCK was in 100% agreement with quantitative PCR with reverse transcription. The assay, which we show is amenable to multiplexed detection in a single lateral-flow strip incorporating an internal control for ribonuclease contamination, should facilitate SARS-CoV-2 detection in settings with limited resources. |COVID-19/*diagnosis/virology[MESH] |CRISPR-Associated Proteins/*genetics[MESH] |Humans[MESH] |Leptotrichia/enzymology[MESH] |Molecular Diagnostic Techniques/*methods[MESH] |Nucleic Acid Amplification Techniques/*methods[MESH] |Pandemics/prevention & control[MESH] |RNA, Viral/*genetics[MESH]Clinical validation of a Cas13-based assay for the detection of SARS-C(epmc).pdf DeepDyve Pubget Overpricing