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10.1016/j.jviromet.2021.114215 http://scihub22266oqcxt.onion/10.1016/j.jviromet.2021.114215 34166701!8215874!34166701     free     free     free Warning: file_get_contents(https://eutils.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&id=34166701&cmd=llinks): Failed to open stream: HTTP request failed! HTTP/1.1 429 Too Many Requests in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 215 Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       J+Virol+Methods 2021 ; 295 (ä): 114215 Nephropedia Template TP {{tp|p=34166701|t=2021. Comparison of SARS-CoV-2 N gene real-time RT-PCR targets and commercially available mastermixes |pdf=|usr=}}{{34166701}} gab.com Text Comparison of SARS-CoV-2 N gene real-time RT-PCR targets and commercially available mastermixes JO: J Virol Methods http://www.kidney.de/pget.php?&tw=1&pmid=34166701 #FOAvip BACKGROUND: This study aimed to evaluate the impact of four different reverse transcription quantitative PCR (RT-qPCR) master mixes on the performance of SARS-CoV-2 diagnostic PCRs using three primer/probe assays targeting the N gene (A, B and C). The dynamic range and lowest detected quantity was determined using a SARS-CoV-2 partial N gene RNA transcript dilution series (100,000-1 copy/mul) and verified using 72 nose and throat swabs, 29 of which tested positive for SARS-CoV-2 RNA. RESULTS: Assay C consistently detected the lowest quantity of partial N gene RNA transcript with all mastermixes. The Takara One Step PrimeScript III RT-PCR Kit mastermix enabled all primer pairs to detect the entire dynamic range evaluated, with the Qiagen Quantifast and Thermofisher TaqPath 1-Step kits also performing well. Sequences from all three primer/probe sets tested in this study (assay A, B and C) have 100 % homology to >/=97 % of the of SARS-CoV-2 sequences available up to 31st December 2020 (n = 291,483 sequences). CONCLUSIONS: This work demonstrates that specific assays (in this case assay C) can perform well in terms of dynamic range and lowest detected quantity regardless of the mastermix used. However we also show that, by choosing the most appropriate mastermix, poorer performing primer pairs are also able to detect all of the template dilutions investigated. This work increases the potential options when choosing assays for SARS-CoV-2 diagnosis and provides solutions to enable them to work with optimal analytical sensitivity. Twit Text FOAVip Comparison of SARS-CoV-2 N gene real-time RT-PCR targets and commercially available mastermixes ????J Virol Methods http://www.kidney.de/pget.php?&tw=1&pmid=34166701 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=34166701 #FOAvip English Wikipedia <ref>{{Cite pmid|34166701}}</ref> Comparison of SARS-CoV-2 N gene real-time RT-PCR targets and commercially available mastermixes #MMPMID34166701 Brown JR; O'Sullivan DM; Shah D; Atkinson L; Pereira RPA; Whale AS; Busby EJ; Huggett JF; Harris K J Virol Methods 2021[Sep]; 295 (ä): 114215 PMID34166701show ga BACKGROUND: This study aimed to evaluate the impact of four different reverse transcription quantitative PCR (RT-qPCR) master mixes on the performance of SARS-CoV-2 diagnostic PCRs using three primer/probe assays targeting the N gene (A, B and C). The dynamic range and lowest detected quantity was determined using a SARS-CoV-2 partial N gene RNA transcript dilution series (100,000-1 copy/mul) and verified using 72 nose and throat swabs, 29 of which tested positive for SARS-CoV-2 RNA. RESULTS: Assay C consistently detected the lowest quantity of partial N gene RNA transcript with all mastermixes. The Takara One Step PrimeScript III RT-PCR Kit mastermix enabled all primer pairs to detect the entire dynamic range evaluated, with the Qiagen Quantifast and Thermofisher TaqPath 1-Step kits also performing well. Sequences from all three primer/probe sets tested in this study (assay A, B and C) have 100 % homology to >/=97 % of the of SARS-CoV-2 sequences available up to 31st December 2020 (n = 291,483 sequences). CONCLUSIONS: This work demonstrates that specific assays (in this case assay C) can perform well in terms of dynamic range and lowest detected quantity regardless of the mastermix used. However we also show that, by choosing the most appropriate mastermix, poorer performing primer pairs are also able to detect all of the template dilutions investigated. This work increases the potential options when choosing assays for SARS-CoV-2 diagnosis and provides solutions to enable them to work with optimal analytical sensitivity. |COVID-19 Nucleic Acid Testing/instrumentation/*methods[MESH] |COVID-19/diagnosis[MESH] |Coronavirus Nucleocapsid Proteins/*genetics[MESH] |DNA Primers/genetics[MESH] |Humans[MESH] |Nose/virology[MESH] |Pharynx/virology[MESH] |Phosphoproteins/genetics[MESH] |RNA, Viral/genetics[MESH] |Reagent Kits, Diagnostic[MESH] |Real-Time Polymerase Chain Reaction[MESH] |Reverse Transcriptase Polymerase Chain Reaction[MESH] |SARS-CoV-2/genetics/*isolation & purification[MESH] |Sensitivity and Specificity[MESH]Comparison of SARS-CoV-2 N gene real-time RT-PCR targets and commercia(epmc).pdf DeepDyve Pubget Overpricing