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10.1007/s42770-020-00347-5 http://scihub22266oqcxt.onion/10.1007/s42770-020-00347-5 32767275!7411266!32767275     free     free     free Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 231.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 265.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 265.2 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       Braz+J+Microbiol 2020 ; 51 (3): 1117-1123 Nephropedia Template TP {{tp|p=32767275|t=2020. Lower cost alternatives for molecular diagnosis of COVID-19: conventional RT-PCR and SYBR Green-based RT-qPCR |pdf=|usr=}}{{32767275}} gab.com Text Lower cost alternatives for molecular diagnosis of COVID-19: conventional RT-PCR and SYBR Green-based RT-qPCR JO: Braz J Microbiol http://www.kidney.de/pget.php?&tw=1&pmid=32767275 #FOAvip In March 2020, WHO declared a pandemic state due to SARS-CoV-2 having spread. TaqMan-based real-time RT-qPCR is currently the gold standard for COVID-19 diagnosis. However, it is a high-cost assay, inaccessible for the majority of laboratories around the world, making it difficult to diagnose on a large scale. The objective of this study was to standardize lower cost molecular methods for SARS-CoV-2 identification. E gene primers previously determined for TaqMan assays by Colman et al. (2020) were adapted in SYBR Green assay and RT-PCR conventional. The cross-reactivity test was performed with 17 positive samples for other respiratory viruses, and the sensibility test was performed with 8 dilutions (10 based) of SARS-CoV-2 isolated and 63 SARS-CoV-2-positive samples. The SYBR Green assays and conventional RT-PCR have not shown amplification of the 17 respiratory samples positives for other viruses. The SYBR Green-based assay was able to detect all 8 dilutions of the isolate. The conventional PCR detected until 10(7) dilution, both assays detected the majority of the 63 samples, 98.42% of positivity in SYBR Green, and 93% in conventional PCR. The average Ct variation between SYBR Green and TaqMan was 1.92 and the highest Ct detected by conventional PCR was 35.98. Both of the proposed assays are less sensitive than the current gold standard; however, our data shows a low sensibility variation, suggesting that these methods could be used by laboratories as a lower cost molecular method for SARS-CoV-2 diagnosis. Twit Text FOAVip Lower cost alternatives for molecular diagnosis of COVID-19: conventional RT-PCR and SYBR Green-based RT-qPCR ????Braz J Microbiol http://www.kidney.de/pget.php?&tw=1&pmid=32767275 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=32767275 #FOAvip English Wikipedia <ref>{{Cite pmid|32767275}}</ref> Lower cost alternatives for molecular diagnosis of COVID-19: conventional RT-PCR and SYBR Green-based RT-qPCR #MMPMID32767275 Dorlass EG; Monteiro CO; Viana AO; Soares CP; Machado RRG; Thomazelli LM; Araujo DB; Leal FB; Candido ED; Telezynski BL; Valerio CA; Chalup VN; Mello R; Almeida FJ; Aguiar AS; Barrientos ACM; Sucupira C; De Paulis M; Safadi MAP; Silva DGBP; Sodre JJM; Soledade MP; Matos SF; Ferreira SR; Pinez CMN; Buonafine CP; Pieroni LNF; Malta FM; Santana RAF; Souza EC; Fock RA; Pinho JRR; Ferreira LCS; Botosso VF; Durigon EL; Oliveira DBL Braz J Microbiol 2020[Sep]; 51 (3): 1117-1123 PMID32767275show ga In March 2020, WHO declared a pandemic state due to SARS-CoV-2 having spread. TaqMan-based real-time RT-qPCR is currently the gold standard for COVID-19 diagnosis. However, it is a high-cost assay, inaccessible for the majority of laboratories around the world, making it difficult to diagnose on a large scale. The objective of this study was to standardize lower cost molecular methods for SARS-CoV-2 identification. E gene primers previously determined for TaqMan assays by Colman et al. (2020) were adapted in SYBR Green assay and RT-PCR conventional. The cross-reactivity test was performed with 17 positive samples for other respiratory viruses, and the sensibility test was performed with 8 dilutions (10 based) of SARS-CoV-2 isolated and 63 SARS-CoV-2-positive samples. The SYBR Green assays and conventional RT-PCR have not shown amplification of the 17 respiratory samples positives for other viruses. The SYBR Green-based assay was able to detect all 8 dilutions of the isolate. The conventional PCR detected until 10(7) dilution, both assays detected the majority of the 63 samples, 98.42% of positivity in SYBR Green, and 93% in conventional PCR. The average Ct variation between SYBR Green and TaqMan was 1.92 and the highest Ct detected by conventional PCR was 35.98. Both of the proposed assays are less sensitive than the current gold standard; however, our data shows a low sensibility variation, suggesting that these methods could be used by laboratories as a lower cost molecular method for SARS-CoV-2 diagnosis. |Adolescent[MESH] |Adult[MESH] |Animals[MESH] |Benzothiazoles[MESH] |Betacoronavirus/genetics/*isolation & purification[MESH] |COVID-19[MESH] |Child[MESH] |Chlorocebus aethiops[MESH] |Coronavirus Infections/*diagnosis/economics[MESH] |Cross Reactions[MESH] |Diamines[MESH] |Fluorescent Dyes/*economics[MESH] |Humans[MESH] |Middle Aged[MESH] |Nasopharynx/virology[MESH] |Organic Chemicals/*economics[MESH] |Oropharynx/virology[MESH] |Pandemics/economics[MESH] |Pneumonia, Viral/*diagnosis/economics[MESH] |Quinolines[MESH] |RNA, Viral/isolation & purification[MESH] |Real-Time Polymerase Chain Reaction/*economics/methods[MESH] |SARS-CoV-2[MESH] |Sensitivity and Specificity[MESH] |Vero Cells[MESH]Lower cost alternatives for molecular diagnosis of COVID-19: conventio(epmc).pdf DeepDyve Pubget Overpricing