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10.1016/j.cca.2020.10.001 http://scihub22266oqcxt.onion/10.1016/j.cca.2020.10.001 33058837!7550048!33058837     free     free     free Deprecated: Implicit conversion from float 215.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 215.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 215.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 215.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 215.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       Clin+Chim+Acta 2020 ; 511 (ä): 149-153 Nephropedia Template TP {{tp|p=33058837|t=2020. Positive control synthesis method for COVID-19 diagnosis by one-step real-time RT-PCR |pdf=|usr=}}{{33058837}} gab.com Text Positive control synthesis method for COVID-19 diagnosis by one-step real-time RT-PCR JO: Clin Chim Acta http://www.kidney.de/pget.php?&tw=1&pmid=33058837 #FOAvip BACKGROUNDS: The coronavirus disease 2019 (COVID-19) pandemic is still ongoing. Real-time reverse transcription polymerase chain reaction (real-time RT-PCR) is regarded as a gold-standard method for the diagnosis of COVID-19. However, unexpected contamination of synthesized positive control samples included in COVID-19 test kits have increased the inconclusiveness of disease interpretation. Therefore, it is important to establish new methods for the preparation of reliable positive controls that are not affected by contamination for the accurate for diagnosis of COVID-19, but it still remains a challenge. METHODS: A new approach for producing synthetic positive controls using synthetic positive template (SPT) oligonucleotides was designed. SPT oligonucleotides contain probe binding and virus-irrelevant regions were used as templates for real-time PCR to evaluate the expression level of SARS-CoV-2 genes (RdRP, E, and N). The limit of detection (LOD) for individual SARS-CoV-2 genes by Ct values with different concentrations of SPT templates and genomic RNAs from SARS-CoV-2 infected samples was determined. RESULTS: LODs with SPT templates were >10(-15) (atto) M for RdRP, 10(-12) (femto) to 10(-13) (100 atto) M for E gene, and 10(-12) to 10(-14) (10 atto) M for N gene, respectively. Real-time RT-PCR assay using serially diluted genomic RNAs prepared from SARS-CoV-2 virus infected cultures showed that picogram quantities of RNAs is resulted in the LOD. The sensitivity of RdRP and E genes based on Ct values was less than that of N gene with this platform. CONCLUSION: This method significantly reduces the risk of false-positive reactions resulting from contamination in the synthesis procedures of positive control materials. Therefore, this approach could be integrated into the currrently available COVID-19 test kits and will provide a general method for preparing positive controls in the diagnosis of emerging RNA virus infections. Twit Text FOAVip Positive control synthesis method for COVID-19 diagnosis by one-step real-time RT-PCR ????Clin Chim Acta http://www.kidney.de/pget.php?&tw=1&pmid=33058837 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=33058837 #FOAvip English Wikipedia <ref>{{Cite pmid|33058837}}</ref> Positive control synthesis method for COVID-19 diagnosis by one-step real-time RT-PCR #MMPMID33058837 Cho H; Jung YH; Cho HB; Kim HT; Kim KS Clin Chim Acta 2020[Dec]; 511 (ä): 149-153 PMID33058837show ga BACKGROUNDS: The coronavirus disease 2019 (COVID-19) pandemic is still ongoing. Real-time reverse transcription polymerase chain reaction (real-time RT-PCR) is regarded as a gold-standard method for the diagnosis of COVID-19. However, unexpected contamination of synthesized positive control samples included in COVID-19 test kits have increased the inconclusiveness of disease interpretation. Therefore, it is important to establish new methods for the preparation of reliable positive controls that are not affected by contamination for the accurate for diagnosis of COVID-19, but it still remains a challenge. METHODS: A new approach for producing synthetic positive controls using synthetic positive template (SPT) oligonucleotides was designed. SPT oligonucleotides contain probe binding and virus-irrelevant regions were used as templates for real-time PCR to evaluate the expression level of SARS-CoV-2 genes (RdRP, E, and N). The limit of detection (LOD) for individual SARS-CoV-2 genes by Ct values with different concentrations of SPT templates and genomic RNAs from SARS-CoV-2 infected samples was determined. RESULTS: LODs with SPT templates were >10(-15) (atto) M for RdRP, 10(-12) (femto) to 10(-13) (100 atto) M for E gene, and 10(-12) to 10(-14) (10 atto) M for N gene, respectively. Real-time RT-PCR assay using serially diluted genomic RNAs prepared from SARS-CoV-2 virus infected cultures showed that picogram quantities of RNAs is resulted in the LOD. The sensitivity of RdRP and E genes based on Ct values was less than that of N gene with this platform. CONCLUSION: This method significantly reduces the risk of false-positive reactions resulting from contamination in the synthesis procedures of positive control materials. Therefore, this approach could be integrated into the currrently available COVID-19 test kits and will provide a general method for preparing positive controls in the diagnosis of emerging RNA virus infections. |COVID-19/*diagnosis/*genetics[MESH] |Humans[MESH] |Reagent Kits, Diagnostic/standards[MESH] |Reverse Transcriptase Polymerase Chain Reaction/*methods/standards[MESH]Positive control synthesis method for COVID-19 diagnosis by one-step r(epmc).pdf DeepDyve Pubget Overpricing