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10.1016/j.jcv.2020.104391 http://scihub22266oqcxt.onion/10.1016/j.jcv.2020.104391 32403008!7192118!32403008     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       J+Clin+Virol 2020 ; 128 (ä): 104391 Nephropedia Template TP {{tp|p=32403008|t=2020. A RT-PCR assay for the detection of coronaviruses from four genera |pdf=|usr=}}{{32403008}} gab.com Text A RT-PCR assay for the detection of coronaviruses from four genera JO: J Clin Virol http://www.kidney.de/pget.php?&tw=1&pmid=32403008 #FOAvip BACKGROUND: During the past two decades, three novel coronaviruses (CoVs) have emerged to cause international human epidemics with severe morbidity. CoVs have also emerged to cause severe epidemics in animals. A better understanding of the natural hosts and genetic diversity of CoVs are needed to help mitigate these threats. OBJECTIVE: To design and evaluate a molecular diagnostic tool for detection and identification of all currently recognized and potentially future emergent CoVs from the Orthocoronavirinae subfamily. STUDY DESIGN AND RESULTS: We designed a semi-nested, reverse transcription RT-PCR assay based upon 38 published genome sequences of human and animal CoVs. We evaluated this assay with 14 human and animal CoVs and 11 other non-CoV respiratory viruses. Through sequencing the assay's target amplicon, the assay correctly identified each of the CoVs; no cross-reactivity with 11 common respiratory viruses was observed. The limits of detection ranged from 4 to 4?x?10(2) copies/reaction, depending on the CoV species tested. To assess the assay's clinical performance, we tested a large panel of previously studied specimens: 192 human respiratory specimens from pneumonia patients, 5 clinical specimens from COVID-19 patients, 81 poultry oral secretion specimens, 109 pig slurry specimens, and 31 aerosol samples from a live bird market. The amplicons of all RT-PCR-positive samples were confirmed by Sanger sequencing. Our assay performed well with all tested specimens across all sample types. CONCLUSIONS: This assay can be used for detection and identification of all previously recognized CoVs, including SARS-CoV-2, and potentially any emergent CoVs in the Orthocoronavirinae subfamily. Twit Text FOAVip A RT-PCR assay for the detection of coronaviruses from four genera ????J Clin Virol http://www.kidney.de/pget.php?&tw=1&pmid=32403008 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=32403008 #FOAvip English Wikipedia <ref>{{Cite pmid|32403008}}</ref> A RT-PCR assay for the detection of coronaviruses from four genera #MMPMID32403008 Xiu L; Binder RA; Alarja NA; Kochek K; Coleman KK; Than ST; Bailey ES; Bui VN; Toh TH; Erdman DD; Gray GC J Clin Virol 2020[Jul]; 128 (ä): 104391 PMID32403008show ga BACKGROUND: During the past two decades, three novel coronaviruses (CoVs) have emerged to cause international human epidemics with severe morbidity. CoVs have also emerged to cause severe epidemics in animals. A better understanding of the natural hosts and genetic diversity of CoVs are needed to help mitigate these threats. OBJECTIVE: To design and evaluate a molecular diagnostic tool for detection and identification of all currently recognized and potentially future emergent CoVs from the Orthocoronavirinae subfamily. STUDY DESIGN AND RESULTS: We designed a semi-nested, reverse transcription RT-PCR assay based upon 38 published genome sequences of human and animal CoVs. We evaluated this assay with 14 human and animal CoVs and 11 other non-CoV respiratory viruses. Through sequencing the assay's target amplicon, the assay correctly identified each of the CoVs; no cross-reactivity with 11 common respiratory viruses was observed. The limits of detection ranged from 4 to 4?x?10(2) copies/reaction, depending on the CoV species tested. To assess the assay's clinical performance, we tested a large panel of previously studied specimens: 192 human respiratory specimens from pneumonia patients, 5 clinical specimens from COVID-19 patients, 81 poultry oral secretion specimens, 109 pig slurry specimens, and 31 aerosol samples from a live bird market. The amplicons of all RT-PCR-positive samples were confirmed by Sanger sequencing. Our assay performed well with all tested specimens across all sample types. CONCLUSIONS: This assay can be used for detection and identification of all previously recognized CoVs, including SARS-CoV-2, and potentially any emergent CoVs in the Orthocoronavirinae subfamily. |Animals[MESH] |Betacoronavirus/genetics/isolation & purification[MESH] |Bird Diseases/*diagnosis/virology[MESH] |Birds[MESH] |COVID-19[MESH] |Coronavirus Infections/*diagnosis/virology[MESH] |Coronavirus/genetics/*isolation & purification[MESH] |Genetic Variation[MESH] |Humans[MESH] |Molecular Diagnostic Techniques/*methods[MESH] |Pandemics[MESH] |Pneumonia, Viral/*diagnosis/virology[MESH] |Reverse Transcriptase Polymerase Chain Reaction/*methods[MESH] |SARS-CoV-2[MESH] |Severe acute respiratory syndrome-related coronavirus/genetics/isolation & purification[MESH] |Swine[MESH]A RT-PCR assay for the detection of coronaviruses from four genera (epmc).pdf DeepDyve Pubget Overpricing