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10.1038/s41598-020-80715-1 http://scihub22266oqcxt.onion/10.1038/s41598-020-80715-1 33436939!7804156!33436939     free     free     free Warning: file_get_contents(https://eutils.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&id=33436939&cmd=llinks): Failed to open stream: HTTP request failed! 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Digital droplet PCR accurately quantifies SARS-CoV-2 viral load from crude lysate without nucleic acid purification |pdf=|usr=}}{{33436939}} gab.com Text Digital droplet PCR accurately quantifies SARS-CoV-2 viral load from crude lysate without nucleic acid purification JO: Sci Rep http://www.kidney.de/pget.php?&tw=1&pmid=33436939 #FOAvip The COVID-19 pandemic caused by the SARS-CoV-2 virus motivates diverse diagnostic approaches due to the novel causative pathogen, incompletely understood clinical sequelae, and limited availability of testing resources. Given the variability in viral load across and within patients, absolute viral load quantification directly from crude lysate is important for diagnosis and surveillance. Here, we investigate the use of digital droplet PCR (ddPCR) for SARS-CoV-2 viral load measurement directly from crude lysate without nucleic acid purification. We demonstrate ddPCR accurately quantifies SARS-CoV-2 standards from purified RNA and multiple sample matrices, including commonly utilized universal transport medium (UTM). In addition, we find ddPCR functions robustly at low input viral copy numbers on nasopharyngeal swab specimens stored in UTM without upfront RNA extraction. We also show ddPCR, but not qPCR, from crude lysate shows high concordance with viral load measurements from purified RNA. Our data suggest ddPCR offers advantages to qPCR for SARS-CoV-2 detection with higher sensitivity and robustness when using crude lysate rather than purified RNA as input. More broadly, digital droplet assays provide a potential method for nucleic acid measurement and infectious disease diagnosis with limited sample processing, underscoring the utility of such techniques in laboratory medicine. Twit Text FOAVip Digital droplet PCR accurately quantifies SARS-CoV-2 viral load from crude lysate without nucleic acid purification ????Sci Rep http://www.kidney.de/pget.php?&tw=1&pmid=33436939 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=33436939 #FOAvip English Wikipedia <ref>{{Cite pmid|33436939}}</ref> Digital droplet PCR accurately quantifies SARS-CoV-2 viral load from crude lysate without nucleic acid purification #MMPMID33436939 Vasudevan HN; Xu P; Servellita V; Miller S; Liu L; Gopez A; Chiu CY; Abate AR Sci Rep 2021[Jan]; 11 (1): 780 PMID33436939show ga The COVID-19 pandemic caused by the SARS-CoV-2 virus motivates diverse diagnostic approaches due to the novel causative pathogen, incompletely understood clinical sequelae, and limited availability of testing resources. Given the variability in viral load across and within patients, absolute viral load quantification directly from crude lysate is important for diagnosis and surveillance. Here, we investigate the use of digital droplet PCR (ddPCR) for SARS-CoV-2 viral load measurement directly from crude lysate without nucleic acid purification. We demonstrate ddPCR accurately quantifies SARS-CoV-2 standards from purified RNA and multiple sample matrices, including commonly utilized universal transport medium (UTM). In addition, we find ddPCR functions robustly at low input viral copy numbers on nasopharyngeal swab specimens stored in UTM without upfront RNA extraction. We also show ddPCR, but not qPCR, from crude lysate shows high concordance with viral load measurements from purified RNA. Our data suggest ddPCR offers advantages to qPCR for SARS-CoV-2 detection with higher sensitivity and robustness when using crude lysate rather than purified RNA as input. More broadly, digital droplet assays provide a potential method for nucleic acid measurement and infectious disease diagnosis with limited sample processing, underscoring the utility of such techniques in laboratory medicine. |*Viral Load[MESH] |COVID-19 Nucleic Acid Testing/*methods/standards[MESH] |COVID-19/diagnosis/*virology[MESH] |Humans[MESH] |Nasal Mucosa/virology[MESH] |RNA, Viral/chemistry/genetics/standards[MESH] |SARS-CoV-2/genetics/isolation & purification/pathogenicity[MESH]Digital droplet PCR accurately quantifies SARS-CoV-2 viral load from c(epmc).pdf DeepDyve Pubget Overpricing