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10.1128/JCM.00958-20 http://scihub22266oqcxt.onion/10.1128/JCM.00958-20 32467359!7383554!32467359     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 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 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 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 263.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       J+Clin+Microbiol 2020 ; 58 (8): ä Nephropedia Template TP {{tp|p=32467359|t=2020. Influence of Different Inactivation Methods on Severe Acute Respiratory Syndrome Coronavirus 2 RNA Copy Number |pdf=|usr=}}{{32467359}} gab.com Text Influence of Different Inactivation Methods on Severe Acute Respiratory Syndrome Coronavirus 2 RNA Copy Number JO: J Clin Microbiol http://www.kidney.de/pget.php?&tw=1&pmid=32467359 #FOAvip The outbreak of coronavirus disease 2019 (COVID-19) has spread across the world and was characterized as a pandemic. To protect medical laboratory personnel from infection, most laboratories inactivate the virus causing COVID-19, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), in clinical samples before testing. However, the effect of inactivation on the detection results remains unknown. Here, we used a digital PCR assay to determine the absolute SARS-CoV-2 RNA copy number in 63 nasopharyngeal swab samples and assess the effect of inactivation methods on viral RNA copy number. Viral inactivation was performed by three different methods: (i) incubation with the TRIzol LS reagent for 10 min at room temperature, (ii) heating in a water bath at 56 degrees C for 30 min, and (iii) high-temperature treatment, including autoclaving at 121 degrees C for 20 min, boiling at 100 degrees C for 20 min, and heating at 80 degrees C for 20 min. Compared to the amount of RNA in the original sample, TRIzol treatment destroyed 47.54% of the nucleocapsid protein (N) gene and 39.85% of open reading frame (ORF) 1ab. For samples treated at 56 degrees C for 30 min, the copy number of the N gene and ORF 1ab was reduced by 48.55% and 56.40%, respectively. The viral RNA copy number dropped by 50 to 66% after heating at 80 degrees C for 20 min. Nearly no viral RNA was detected after autoclaving at 121 degrees C or boiling at 100 degrees C for 20 min. These results indicate that inactivation reduced the quantity of detectable viral RNA and may cause false-negative results, especially in weakly positive cases. Thus, use of the TRIzol reagent rather than heat inactivation is recommended for sample inactivation, as the TRIzol reagent had the least effect on the RNA copy number among the tested methods. Twit Text FOAVip Influence of Different Inactivation Methods on Severe Acute Respiratory Syndrome Coronavirus 2 RNA Copy Number ????J Clin Microbiol http://www.kidney.de/pget.php?&tw=1&pmid=32467359 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=32467359 #FOAvip English Wikipedia <ref>{{Cite pmid|32467359}}</ref> Influence of Different Inactivation Methods on Severe Acute Respiratory Syndrome Coronavirus 2 RNA Copy Number #MMPMID32467359 Chen H; Wu R; Xing Y; Du Q; Xue Z; Xi Y; Yang Y; Deng Y; Han Y; Li K; Luan Y; Zhang Y; Wei X; Yu T; Li H; Zhu L; Su S; Lian H; Lu L; Tan C; Zheng H; Chen B; Yu P; Guo Y; Ma C J Clin Microbiol 2020[Jul]; 58 (8): ä PMID32467359show ga The outbreak of coronavirus disease 2019 (COVID-19) has spread across the world and was characterized as a pandemic. To protect medical laboratory personnel from infection, most laboratories inactivate the virus causing COVID-19, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), in clinical samples before testing. However, the effect of inactivation on the detection results remains unknown. Here, we used a digital PCR assay to determine the absolute SARS-CoV-2 RNA copy number in 63 nasopharyngeal swab samples and assess the effect of inactivation methods on viral RNA copy number. Viral inactivation was performed by three different methods: (i) incubation with the TRIzol LS reagent for 10 min at room temperature, (ii) heating in a water bath at 56 degrees C for 30 min, and (iii) high-temperature treatment, including autoclaving at 121 degrees C for 20 min, boiling at 100 degrees C for 20 min, and heating at 80 degrees C for 20 min. Compared to the amount of RNA in the original sample, TRIzol treatment destroyed 47.54% of the nucleocapsid protein (N) gene and 39.85% of open reading frame (ORF) 1ab. For samples treated at 56 degrees C for 30 min, the copy number of the N gene and ORF 1ab was reduced by 48.55% and 56.40%, respectively. The viral RNA copy number dropped by 50 to 66% after heating at 80 degrees C for 20 min. Nearly no viral RNA was detected after autoclaving at 121 degrees C or boiling at 100 degrees C for 20 min. These results indicate that inactivation reduced the quantity of detectable viral RNA and may cause false-negative results, especially in weakly positive cases. Thus, use of the TRIzol reagent rather than heat inactivation is recommended for sample inactivation, as the TRIzol reagent had the least effect on the RNA copy number among the tested methods. |Adolescent[MESH] |Adult[MESH] |Aged[MESH] |Aged, 80 and over[MESH] |Betacoronavirus/*drug effects/*radiation effects[MESH] |Disinfectants[MESH] |Disinfection/*methods[MESH] |Female[MESH] |Gene Dosage[MESH] |Hot Temperature[MESH] |Humans[MESH] |Male[MESH] |Middle Aged[MESH] |Polymerase Chain Reaction[MESH] |RNA, Viral/*analysis/genetics[MESH] |SARS-CoV-2[MESH] |Specimen Handling/*methods[MESH] |Virus Inactivation/*drug effects/*radiation effects[MESH]Influence of Different Inactivation Methods on Severe Acute Respirator(epmc).pdf DeepDyve Pubget Overpricing