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10.3855/jidc.12726 http://scihub22266oqcxt.onion/10.3855/jidc.12726 32794453!ä!32794453 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 267.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 267.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       J+Infect+Dev+Ctries 2020 ; 14 (7): 679-684 Nephropedia Template TP {{tp|p=32794453|t=2020. One-step nested RT-PCR for COVID-19 detection: A flexible, locally developed test for SARS-CoV2 nucleic acid detection |pdf=|usr=}}{{32794453}} gab.com Text One-step nested RT-PCR for COVID-19 detection: A flexible, locally developed test for SARS-CoV2 nucleic acid detection JO: J Infect Dev Ctries http://www.kidney.de/pget.php?&tw=1&pmid=32794453 #FOAvip INTRODUCTION: Due to the coronavirus pandemic, identifying the infected individuals has become key to limiting its spread. Virus nucleic acid real-time RT-PCR testing has become the current standard diagnostic method but high demand could lead to shortages. Therefore, we propose a detection strategy using a one-step nested RT-PCR. METHODOLOGY: The nucleotide region in the ORF1ab gene that has the greatest differences between the human coronavirus and the bat coronavirus was selected. Primers were designed after that sequence. All diagnostic primers are species-specific since the 3 end of the sequence differs from that of other species. A primer set also creates a synthetic positive control. Amplified products were seen in a 2.5% agarose gel, as well as in an SYBR Green-Based Real-Time RT-PCR. RESULTS: Amplification was achieved for the positive control and specific regions in both techniques. CONCLUSIONS: This new technique is flexible and easy to implement. It does not require a real-time thermocycler and can be interpreted in agarose gels, as well as adapted to quantify the viral genome. It has the advantage that if the coronavirus mutates in one of the key amplification nucleotides, at least one pair can still amplify, thanks to the four diagnostic primers. Twit Text FOAVip One-step nested RT-PCR for COVID-19 detection: A flexible, locally developed test for SARS-CoV2 nucleic acid detection ????J Infect Dev Ctries http://www.kidney.de/pget.php?&tw=1&pmid=32794453 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=32794453 #FOAvip English Wikipedia <ref>{{Cite pmid|32794453}}</ref> One-step nested RT-PCR for COVID-19 detection: A flexible, locally developed test for SARS-CoV2 nucleic acid detection #MMPMID32794453 Meza-Robles C; Barajas-Saucedo CE; Tiburcio-Jimenez D; Mokay-Ramirez KA; Melnikov V; Rodriguez-Sanchez IP; Martinez-Fierro ML; Garza-Veloz I; Zaizar-Fregoso SA; Guzman-Esquivel J; Ramirez-Flores M; Newton-Sanchez OA; Espinoza-Gomez F; Delgado-Enciso OG; Centeno-Ramirez AS; Delgado-Enciso I J Infect Dev Ctries 2020[Jul]; 14 (7): 679-684 PMID32794453show ga INTRODUCTION: Due to the coronavirus pandemic, identifying the infected individuals has become key to limiting its spread. Virus nucleic acid real-time RT-PCR testing has become the current standard diagnostic method but high demand could lead to shortages. Therefore, we propose a detection strategy using a one-step nested RT-PCR. METHODOLOGY: The nucleotide region in the ORF1ab gene that has the greatest differences between the human coronavirus and the bat coronavirus was selected. Primers were designed after that sequence. All diagnostic primers are species-specific since the 3 end of the sequence differs from that of other species. A primer set also creates a synthetic positive control. Amplified products were seen in a 2.5% agarose gel, as well as in an SYBR Green-Based Real-Time RT-PCR. RESULTS: Amplification was achieved for the positive control and specific regions in both techniques. CONCLUSIONS: This new technique is flexible and easy to implement. It does not require a real-time thermocycler and can be interpreted in agarose gels, as well as adapted to quantify the viral genome. It has the advantage that if the coronavirus mutates in one of the key amplification nucleotides, at least one pair can still amplify, thanks to the four diagnostic primers. |*Betacoronavirus[MESH] |COVID-19[MESH] |COVID-19 Testing[MESH] |COVID-19 Vaccines[MESH] |Clinical Laboratory Techniques[MESH] |Coronavirus Infections/*diagnosis[MESH] |Humans[MESH] |Pandemics[MESH] |Pneumonia, Viral/*diagnosis[MESH] |RNA, Viral/*analysis[MESH] |Real-Time Polymerase Chain Reaction/*methods[MESH]One-step nested RT-PCR for COVID-19 detection: A flexible, locally dev(epmc).pdf DeepDyve Pubget Overpricing