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10.1007/s10096-020-04076-3 http://scihub22266oqcxt.onion/10.1007/s10096-020-04076-3 33104899!7587514!33104899     free     free     free Deprecated: Implicit conversion from float 219.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 219.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 219.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 219.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 219.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       Eur+J+Clin+Microbiol+Infect+Dis 2021 ; 40 (4): 807-813 Nephropedia Template TP {{tp|p=33104899|t=2021. Sensitive detection and quantification of SARS-CoV-2 by multiplex droplet digital RT-PCR |pdf=|usr=}}{{33104899}} gab.com Text Sensitive detection and quantification of SARS-CoV-2 by multiplex droplet digital RT-PCR JO: Eur J Clin Microbiol Infect Dis http://www.kidney.de/pget.php?&tw=1&pmid=33104899 #FOAvip The purpose of this study is to develop a one-step droplet digital RT-PCR (RT-ddPCR) multiplex assay that allows for sensitive quantification of SARS-CoV-2 RNA with respect to human-derived RNA and could be used for screening and monitoring of Covid-19 patients. A one-step RT-ddPCR multiplex assay was developed for simultaneous detection of SARS-CoV-2 E, RdRp and N viral RNA, and human Rpp30 DNA and GUSB mRNA, for internal nucleic acid (NA) extraction and RT-PCR control. Dilution series of viral RNA transcripts were prepared in water and total NA extract of Covid-19-negative patients. As reference assay, an E-GUSB duplex RT-PCR was used. GUSB mRNA detection was used to set validity criteria to assure viral RNA and RT-PCR assay quality and to enable quantification of SARS-CoV-2 RNA. In a background of at least 100 GUSB mRNA copies, 5 copies of viral RNA are reliably detectable and 10 copies viral RNA copies are reliably quantifiable. It was found that assay sensitivity of the RT-ddPCR was not affected by the total NA background while assay sensitivity of the gold standard RT-PCR assay is drastically decreased when SARS-CoV-2 copies were detected in a background of total NA extract compared with water. The present study describes a robust and sensitive one-step ddRT-PCR multiplex assay for reliable quantification of SARS-CoV-2 RNA. By determining the fractional abundance of viral RNA with respect to a human housekeeping gene, viral loads from different samples can be compared, what could be used to investigate the infectiveness and to monitor Covid-19 patients. Twit Text FOAVip Sensitive detection and quantification of SARS-CoV-2 by multiplex droplet digital RT-PCR ????Eur J Clin Microbiol Infect Dis http://www.kidney.de/pget.php?&tw=1&pmid=33104899 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=33104899 #FOAvip English Wikipedia <ref>{{Cite pmid|33104899}}</ref> Sensitive detection and quantification of SARS-CoV-2 by multiplex droplet digital RT-PCR #MMPMID33104899 de Kock R; Baselmans M; Scharnhorst V; Deiman B Eur J Clin Microbiol Infect Dis 2021[Apr]; 40 (4): 807-813 PMID33104899show ga The purpose of this study is to develop a one-step droplet digital RT-PCR (RT-ddPCR) multiplex assay that allows for sensitive quantification of SARS-CoV-2 RNA with respect to human-derived RNA and could be used for screening and monitoring of Covid-19 patients. A one-step RT-ddPCR multiplex assay was developed for simultaneous detection of SARS-CoV-2 E, RdRp and N viral RNA, and human Rpp30 DNA and GUSB mRNA, for internal nucleic acid (NA) extraction and RT-PCR control. Dilution series of viral RNA transcripts were prepared in water and total NA extract of Covid-19-negative patients. As reference assay, an E-GUSB duplex RT-PCR was used. GUSB mRNA detection was used to set validity criteria to assure viral RNA and RT-PCR assay quality and to enable quantification of SARS-CoV-2 RNA. In a background of at least 100 GUSB mRNA copies, 5 copies of viral RNA are reliably detectable and 10 copies viral RNA copies are reliably quantifiable. It was found that assay sensitivity of the RT-ddPCR was not affected by the total NA background while assay sensitivity of the gold standard RT-PCR assay is drastically decreased when SARS-CoV-2 copies were detected in a background of total NA extract compared with water. The present study describes a robust and sensitive one-step ddRT-PCR multiplex assay for reliable quantification of SARS-CoV-2 RNA. By determining the fractional abundance of viral RNA with respect to a human housekeeping gene, viral loads from different samples can be compared, what could be used to investigate the infectiveness and to monitor Covid-19 patients. |Autoantigens/genetics[MESH] |COVID-19 Nucleic Acid Testing/*methods[MESH] |COVID-19/*diagnosis[MESH] |Coronavirus Envelope Proteins/genetics[MESH] |Coronavirus Nucleocapsid Proteins/genetics[MESH] |Coronavirus RNA-Dependent RNA Polymerase/genetics[MESH] |DNA/*analysis[MESH] |Genes, Essential[MESH] |Glucuronidase/genetics[MESH] |Humans[MESH] |Multiplex Polymerase Chain Reaction/*methods[MESH] |Phosphoproteins/genetics[MESH] |RNA, Messenger/*analysis[MESH] |RNA, Viral/*analysis[MESH] |Real-Time Polymerase Chain Reaction[MESH] |Reverse Transcriptase Polymerase Chain Reaction/*methods[MESH] |Ribonuclease P/genetics[MESH] |SARS-CoV-2[MESH]Sensitive detection and quantification of SARS-CoV-2 by multiplex drop(epmc).pdf DeepDyve Pubget Overpricing