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10.1016/j.jcv.2020.104579 http://scihub22266oqcxt.onion/10.1016/j.jcv.2020.104579 32795959!7405857!32795959     free     free     free Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 211.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       J+Clin+Virol 2020 ; 130 (ä): 104579 Nephropedia Template TP {{tp|p=32795959|t=2020. Extraction-free SARS-CoV-2 detection by rapid RT-qPCR universal for all primary respiratory materials |pdf=|usr=}}{{32795959}} gab.com Text Extraction-free SARS-CoV-2 detection by rapid RT-qPCR universal for all primary respiratory materials JO: J Clin Virol http://www.kidney.de/pget.php?&tw=1&pmid=32795959 #FOAvip BACKGROUND: Fast and reliable detection of SARS-CoV-2 is crucial for efficient control of the COVID-19 pandemic. Due to the high demand for SARS-CoV-2 testing there is a worldwide shortage of RNA extraction reagents. Therefore, extraction-free RT-qPCR protocols are urgently needed. OBJECTIVES: To establish a rapid RT-qPCR protocol for the detection of SARS-CoV-2 without the need of RNA extraction suitable for all respiratory materials. MATERIAL AND METHODS: Different SARS-CoV-2 positive respiratory materials from our routine laboratory were used as crude material after heat inactivation in direct RT-qPCR with the PrimeDirect Probe RT-qPCR Mix (TaKaRa). SARS-CoV-2 was detected using novel primers targeted to the E-gene. RESULTS: The protocol for the detection of SARS-CoV-2 in crude material used a prepared frozen-PCR mix with optimized primers and 5 mul of fresh, undiluted and pre-analytically heat inactivated respiratory material. For validation, 91 respiratory samples were analyzed in direct comparison to classical RNA-based RT-qPCR. Overall 81.3 % of the samples were detected in both assays with a strong correlation between both Ct values (r = 0.8492, p < 0.0001). The SARS-CoV-2 detection rate by direct RT-qPCR was 95.8 % for Ct values <35. All negative samples were characterized by low viral loads (Ct >35) and/or long storage times before sample processing. CONCLUSION: Direct RT-qPCR is a suitable alternative to classical RNA RT-qPCR, provided that only fresh samples (storage <1 week) are used. RNA extraction should be considered if samples have longer storage times or if PCR inhibition is observed. In summary, this protocol is fast, inexpensive and suitable for all respiratory materials. Twit Text FOAVip Extraction-free SARS-CoV-2 detection by rapid RT-qPCR universal for all primary respiratory materials ????J Clin Virol http://www.kidney.de/pget.php?&tw=1&pmid=32795959 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=32795959 #FOAvip English Wikipedia <ref>{{Cite pmid|32795959}}</ref> Extraction-free SARS-CoV-2 detection by rapid RT-qPCR universal for all primary respiratory materials #MMPMID32795959 Lubke N; Senff T; Scherger S; Hauka S; Andree M; Adams O; Timm J; Walker A J Clin Virol 2020[Sep]; 130 (ä): 104579 PMID32795959show ga BACKGROUND: Fast and reliable detection of SARS-CoV-2 is crucial for efficient control of the COVID-19 pandemic. Due to the high demand for SARS-CoV-2 testing there is a worldwide shortage of RNA extraction reagents. Therefore, extraction-free RT-qPCR protocols are urgently needed. OBJECTIVES: To establish a rapid RT-qPCR protocol for the detection of SARS-CoV-2 without the need of RNA extraction suitable for all respiratory materials. MATERIAL AND METHODS: Different SARS-CoV-2 positive respiratory materials from our routine laboratory were used as crude material after heat inactivation in direct RT-qPCR with the PrimeDirect Probe RT-qPCR Mix (TaKaRa). SARS-CoV-2 was detected using novel primers targeted to the E-gene. RESULTS: The protocol for the detection of SARS-CoV-2 in crude material used a prepared frozen-PCR mix with optimized primers and 5 mul of fresh, undiluted and pre-analytically heat inactivated respiratory material. For validation, 91 respiratory samples were analyzed in direct comparison to classical RNA-based RT-qPCR. Overall 81.3 % of the samples were detected in both assays with a strong correlation between both Ct values (r = 0.8492, p < 0.0001). The SARS-CoV-2 detection rate by direct RT-qPCR was 95.8 % for Ct values <35. All negative samples were characterized by low viral loads (Ct >35) and/or long storage times before sample processing. CONCLUSION: Direct RT-qPCR is a suitable alternative to classical RNA RT-qPCR, provided that only fresh samples (storage <1 week) are used. RNA extraction should be considered if samples have longer storage times or if PCR inhibition is observed. In summary, this protocol is fast, inexpensive and suitable for all respiratory materials. |Betacoronavirus[MESH] |COVID-19[MESH] |COVID-19 Testing[MESH] |Clinical Laboratory Techniques/*methods[MESH] |Coronavirus Infections/*diagnosis/virology[MESH] |DNA Primers/genetics[MESH] |Humans[MESH] |Pandemics[MESH] |Pneumonia, Viral/*diagnosis/virology[MESH] |RNA, Viral/analysis[MESH] |Real-Time Polymerase Chain Reaction/*methods[MESH] |Respiratory System/*virology[MESH] |SARS-CoV-2[MESH] |Sensitivity and Specificity[MESH] |Specimen Handling/*methods[MESH]Extraction-free SARS-CoV-2 detection by rapid RT-qPCR universal for al(epmc).pdf DeepDyve Pubget Overpricing