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10.1016/j.ijid.2021.01.001 http://scihub22266oqcxt.onion/10.1016/j.ijid.2021.01.001 33434663!7831874!33434663     free     free     free Warning: file_get_contents(https://eutils.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&id=33434663&cmd=llinks): Failed to open stream: HTTP request failed! 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Development of a quantitative one-step multiplex RT-qPCR assay for the detection of SARS-CoV-2 in a biological matrix |pdf=|usr=}}{{33434663}} gab.com Text Development of a quantitative one-step multiplex RT-qPCR assay for the detection of SARS-CoV-2 in a biological matrix JO: Int J Infect Dis http://www.kidney.de/pget.php?&tw=1&pmid=33434663 #FOAvip INTRODUCTION: Coronavirus disease-2019 (COVID-19) is a disease caused by Severe Acute Respiratory Syndrome Virus 2 (SARS-CoV-2) that emerged in China in late 2019. The rapid viral spread has made the disease a public health emergency of worldwide concern. The gold standard for diagnosing SARS-CoV-2 is reverse transcription followed by qualitative real-time polymerase chain reaction (RT-qPCR); however, the role of viral load quantification has not been thoroughly investigated yet. OBJECTIVE: The aim of this study was to develop a high-precision quantitative one-step RT-qPCR reaction using the association of the viral target and the human target in the same reaction. METHODS: The assay standardization involved the absolute quantification method, with serial dilutions of a plasmid with the N gene in a biological matrix to build a standard curve. RESULTS AND DISCUSSION: The results demonstrated the possibility of quantifying as few as 2.5 copies/reaction and an analysis of 244 patients with known results selected by cross-section that revealed 100% agreement with a qualitative RT-qPCR assay registered by Anvisa. In this population, it was possible to quantify patients with between 2.59 and 3.5 x 10(7) copies per reaction and negative patients continued to indicate the same result. CONCLUSION: This assay can be a useful tool for a proper patient management, because the level and duration of viral replication are important factors to assess the risk of transmission and to guide decisions regarding the isolation and release of patients; an accurate diagnosis is critical information, whereas the current COVID-19 pandemic represents the biggest current global health problem. Twit Text FOAVip Development of a quantitative one-step multiplex RT-qPCR assay for the detection of SARS-CoV-2 in a biological matrix ????Int J Infect Dis http://www.kidney.de/pget.php?&tw=1&pmid=33434663 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=33434663 #FOAvip English Wikipedia <ref>{{Cite pmid|33434663}}</ref> Development of a quantitative one-step multiplex RT-qPCR assay for the detection of SARS-CoV-2 in a biological matrix #MMPMID33434663 Queiroz JADS; Rampazzo RCP; Filho EBDS; Oliveira GS; Oliveira SDC; Souza LFB; Pereira SDS; Rodrigues MMS; Maia ACS; da Silva CC; Mendonca ALFM; Lugtenburg CAB; Aguiar FAA; Rodrigues RSS; Santos CHN; Guimaraes APDS; Maximo FR; Santos AOD; Krieger MA; Salcedo JMV; Dall'Acqua DSV Int J Infect Dis 2021[Mar]; 104 (ä): 373-378 PMID33434663show ga INTRODUCTION: Coronavirus disease-2019 (COVID-19) is a disease caused by Severe Acute Respiratory Syndrome Virus 2 (SARS-CoV-2) that emerged in China in late 2019. The rapid viral spread has made the disease a public health emergency of worldwide concern. The gold standard for diagnosing SARS-CoV-2 is reverse transcription followed by qualitative real-time polymerase chain reaction (RT-qPCR); however, the role of viral load quantification has not been thoroughly investigated yet. OBJECTIVE: The aim of this study was to develop a high-precision quantitative one-step RT-qPCR reaction using the association of the viral target and the human target in the same reaction. METHODS: The assay standardization involved the absolute quantification method, with serial dilutions of a plasmid with the N gene in a biological matrix to build a standard curve. RESULTS AND DISCUSSION: The results demonstrated the possibility of quantifying as few as 2.5 copies/reaction and an analysis of 244 patients with known results selected by cross-section that revealed 100% agreement with a qualitative RT-qPCR assay registered by Anvisa. In this population, it was possible to quantify patients with between 2.59 and 3.5 x 10(7) copies per reaction and negative patients continued to indicate the same result. CONCLUSION: This assay can be a useful tool for a proper patient management, because the level and duration of viral replication are important factors to assess the risk of transmission and to guide decisions regarding the isolation and release of patients; an accurate diagnosis is critical information, whereas the current COVID-19 pandemic represents the biggest current global health problem. |Adolescent[MESH] |Adult[MESH] |Aged[MESH] |Aged, 80 and over[MESH] |COVID-19 Nucleic Acid Testing/*methods[MESH] |COVID-19/*diagnosis/virology[MESH] |Child[MESH] |Child, Preschool[MESH] |Female[MESH] |Humans[MESH] |Infant[MESH] |Male[MESH] |Middle Aged[MESH] |Multiplex Polymerase Chain Reaction[MESH] |Real-Time Polymerase Chain Reaction[MESH] |Reference Standards[MESH] |SARS-CoV-2/*isolation & purification[MESH] |Sensitivity and Specificity[MESH] |Viral Load[MESH]Development of a quantitative one-step multiplex RT-qPCR assay for the(epmc).pdf DeepDyve Pubget Overpricing