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10.1016/j.cmi.2020.04.001 http://scihub22266oqcxt.onion/10.1016/j.cmi.2020.04.001 32276116!7144850!32276116     free     free     free Deprecated: Implicit conversion from float 235.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 235.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 235.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 235.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 235.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 269.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 269.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 269.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 269.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       Clin+Microbiol+Infect 2020 ; 26 (6): 773-779 Nephropedia Template TP {{tp|p=32276116|t=2020. Rapid and visual detection of 2019 novel coronavirus (SARS-CoV-2) by a reverse transcription loop-mediated isothermal amplification assay |pdf=|usr=}}{{32276116}} gab.com Text Rapid and visual detection of 2019 novel coronavirus (SARS-CoV-2) by a reverse transcription loop-mediated isothermal amplification assay JO: Clin Microbiol Infect http://www.kidney.de/pget.php?&tw=1&pmid=32276116 #FOAvip OBJECTIVE: To evaluate a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and compare it with RT-PCR. METHODS: We designed primers specific to the orf1ab and S genes of SARS-CoV-2. Total viral RNA was extracted using the QIAamp Viral RNA Mini Kit. We optimized the RT-LAMP assay, and evaluated it for its sensitivity and specificity of detection using real-time turbidity monitoring and visual observation. RESULTS: The primer sets orf1ab-4 and S-123 amplified the genes in the shortest times, the mean (+/-SD) times were 18 +/- 1.32 min and 20 +/- 1.80 min, respectively, and 63 degrees C was the optimum reaction temperature. The sensitivities were 2 x 10(1) copies and 2 x 10(2) copies per reaction with primer sets orf1ab-4 and S-123, respectively. This assay showed no cross-reactivity with 60 other respiratory pathogens. To describe the availability of this method in clinical diagnosis, we collected 130 specimens from patients with clinically suspected SARS-CoV-2 infection. Among them, 58 were confirmed to be positive and 72 were negative by RT-LAMP. The sensitivity was 100% (95% CI 92.3%-100%), specificity 100% (95% CI 93.7%-100%). This assay detected SARS-CoV-2 in a mean (+/-SD) time of 26.28 +/- 4.48 min and the results can be identified with visual observation. CONCLUSION: These results demonstrate that we developed a rapid, simple, specific and sensitive RT-LAMP assay for SARS-CoV-2 detection among clinical samples. It will be a powerful tool for SARS-CoV-2 identification, and for monitoring suspected patients, close contacts and high-risk groups. Twit Text FOAVip Rapid and visual detection of 2019 novel coronavirus (SARS-CoV-2) by a reverse transcription loop-mediated isothermal amplification assay ????Clin Microbiol Infect http://www.kidney.de/pget.php?&tw=1&pmid=32276116 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=32276116 #FOAvip English Wikipedia <ref>{{Cite pmid|32276116}}</ref> Rapid and visual detection of 2019 novel coronavirus (SARS-CoV-2) by a reverse transcription loop-mediated isothermal amplification assay #MMPMID32276116 Yan C; Cui J; Huang L; Du B; Chen L; Xue G; Li S; Zhang W; Zhao L; Sun Y; Yao H; Li N; Zhao H; Feng Y; Liu S; Zhang Q; Liu D; Yuan J Clin Microbiol Infect 2020[Jun]; 26 (6): 773-779 PMID32276116show ga OBJECTIVE: To evaluate a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), and compare it with RT-PCR. METHODS: We designed primers specific to the orf1ab and S genes of SARS-CoV-2. Total viral RNA was extracted using the QIAamp Viral RNA Mini Kit. We optimized the RT-LAMP assay, and evaluated it for its sensitivity and specificity of detection using real-time turbidity monitoring and visual observation. RESULTS: The primer sets orf1ab-4 and S-123 amplified the genes in the shortest times, the mean (+/-SD) times were 18 +/- 1.32 min and 20 +/- 1.80 min, respectively, and 63 degrees C was the optimum reaction temperature. The sensitivities were 2 x 10(1) copies and 2 x 10(2) copies per reaction with primer sets orf1ab-4 and S-123, respectively. This assay showed no cross-reactivity with 60 other respiratory pathogens. To describe the availability of this method in clinical diagnosis, we collected 130 specimens from patients with clinically suspected SARS-CoV-2 infection. Among them, 58 were confirmed to be positive and 72 were negative by RT-LAMP. The sensitivity was 100% (95% CI 92.3%-100%), specificity 100% (95% CI 93.7%-100%). This assay detected SARS-CoV-2 in a mean (+/-SD) time of 26.28 +/- 4.48 min and the results can be identified with visual observation. CONCLUSION: These results demonstrate that we developed a rapid, simple, specific and sensitive RT-LAMP assay for SARS-CoV-2 detection among clinical samples. It will be a powerful tool for SARS-CoV-2 identification, and for monitoring suspected patients, close contacts and high-risk groups. |*Real-Time Polymerase Chain Reaction[MESH] |Betacoronavirus/genetics/*isolation & purification[MESH] |COVID-19[MESH] |COVID-19 Testing[MESH] |COVID-19 Vaccines[MESH] |Clinical Laboratory Techniques[MESH] |Coronavirus Infections/*diagnosis[MESH] |Humans[MESH] |Molecular Diagnostic Techniques/*methods[MESH] |Nucleic Acid Amplification Techniques/*methods[MESH] |Pandemics[MESH] |Pneumonia, Viral/*diagnosis[MESH] |Polyproteins[MESH] |SARS-CoV-2[MESH] |Sensitivity and Specificity[MESH] |Spike Glycoprotein, Coronavirus/analysis[MESH]Rapid and visual detection of 2019 novel coronavirus (SARS-CoV-2) by a(epmc).pdf DeepDyve Pubget Overpricing