Use my Search Websuite to scan PubMed, PMCentral, Journal Hosts and Journal Archives, FullText. Kick-your-searchterm to multiple Engines kick-your-query now !>

A dictionary by aggregated review articles of nephrology, medicine and the life sciences Your one-stop-run pathway from word to the immediate pdf of peer-reviewed on-topic knowledge.

10.1186/s13059-021-02387-y http://scihub22266oqcxt.onion/10.1186/s13059-021-02387-y 34082799!8173101!34082799     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       Genome+Biol 2021 ; 22 (1): 169 Nephropedia Template TP {{tp|p=34082799|t=2021. Detection of SARS-CoV-2 RNA using RT-LAMP and molecular beacons |pdf=|usr=}}{{34082799}} gab.com Text Detection of SARS-CoV-2 RNA using RT-LAMP and molecular beacons JO: Genome Biol http://www.kidney.de/pget.php?&tw=1&pmid=34082799 #FOAvip BACKGROUND: Rapid spread of SARS-CoV-2 has led to a global pandemic, resulting in the need for rapid assays to allow diagnosis and prevention of transmission. Reverse transcription-polymerase chain reaction (RT-PCR) provides a gold standard assay for SARS-CoV-2 RNA, but instrument costs are high and supply chains are potentially fragile, motivating interest in additional assay methods. Reverse transcription and loop-mediated isothermal amplification (RT-LAMP) provides an alternative that uses orthogonal and often less expensive reagents without the need for thermocyclers. The presence of SARS-CoV-2 RNA is typically detected using dyes to report bulk amplification of DNA; however, a common artifact is nonspecific DNA amplification, which complicates detection. RESULTS: Here we describe the design and testing of molecular beacons, which allow sequence-specific detection of SARS-CoV-2 genomes with improved discrimination in simple reaction mixtures. To optimize beacons for RT-LAMP, multiple locked nucleic acid monomers were incorporated to elevate melting temperatures. We also show how beacons with different fluorescent labels can allow convenient multiplex detection of several amplicons in "single pot" reactions, including incorporation of a human RNA LAMP-BEAC assay to confirm sample integrity. Comparison of LAMP-BEAC and RT-qPCR on clinical saliva samples showed good concordance between assays. To facilitate implementation, we developed custom polymerases for LAMP-BEAC and inexpensive purification procedures, which also facilitates increasing sensitivity by increasing reaction volumes. CONCLUSIONS: LAMP-BEAC thus provides an affordable and simple SARS-CoV-2 RNA assay suitable for population screening; implementation of the assay has allowed robust screening of thousands of saliva samples per week. Twit Text FOAVip Detection of SARS-CoV-2 RNA using RT-LAMP and molecular beacons ????Genome Biol http://www.kidney.de/pget.php?&tw=1&pmid=34082799 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=34082799 #FOAvip English Wikipedia <ref>{{Cite pmid|34082799}}</ref> Detection of SARS-CoV-2 RNA using RT-LAMP and molecular beacons #MMPMID34082799 Sherrill-Mix S; Hwang Y; Roche AM; Glascock A; Weiss SR; Li Y; Haddad L; Deraska P; Monahan C; Kromer A; Graham-Wooten J; Taylor LJ; Abella BS; Ganguly A; Collman RG; Van Duyne GD; Bushman FD Genome Biol 2021[Jun]; 22 (1): 169 PMID34082799show ga BACKGROUND: Rapid spread of SARS-CoV-2 has led to a global pandemic, resulting in the need for rapid assays to allow diagnosis and prevention of transmission. Reverse transcription-polymerase chain reaction (RT-PCR) provides a gold standard assay for SARS-CoV-2 RNA, but instrument costs are high and supply chains are potentially fragile, motivating interest in additional assay methods. Reverse transcription and loop-mediated isothermal amplification (RT-LAMP) provides an alternative that uses orthogonal and often less expensive reagents without the need for thermocyclers. The presence of SARS-CoV-2 RNA is typically detected using dyes to report bulk amplification of DNA; however, a common artifact is nonspecific DNA amplification, which complicates detection. RESULTS: Here we describe the design and testing of molecular beacons, which allow sequence-specific detection of SARS-CoV-2 genomes with improved discrimination in simple reaction mixtures. To optimize beacons for RT-LAMP, multiple locked nucleic acid monomers were incorporated to elevate melting temperatures. We also show how beacons with different fluorescent labels can allow convenient multiplex detection of several amplicons in "single pot" reactions, including incorporation of a human RNA LAMP-BEAC assay to confirm sample integrity. Comparison of LAMP-BEAC and RT-qPCR on clinical saliva samples showed good concordance between assays. To facilitate implementation, we developed custom polymerases for LAMP-BEAC and inexpensive purification procedures, which also facilitates increasing sensitivity by increasing reaction volumes. CONCLUSIONS: LAMP-BEAC thus provides an affordable and simple SARS-CoV-2 RNA assay suitable for population screening; implementation of the assay has allowed robust screening of thousands of saliva samples per week. |COVID-19 Testing[MESH] |COVID-19/*diagnosis[MESH] |Humans[MESH] |Molecular Diagnostic Techniques[MESH] |Nucleic Acid Amplification Techniques[MESH] |Nucleic Acid Probes/genetics[MESH] |RNA, Viral/*genetics[MESH] |SARS-CoV-2/genetics/*isolation & purification[MESH] |Saliva/virology[MESH]Detection of SARS-CoV-2 RNA using RT-LAMP and molecular beacons (epmc).pdf DeepDyve Pubget Overpricing