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10.1128/JCM.00963-19 http://scihub22266oqcxt.onion/10.1128/JCM.00963-19 31666364!6935926!31666364     free     free     free Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534 Deprecated: Implicit conversion from float 233.6 to int loses precision in C:\Inetpub\vhosts\kidney.de\httpdocs\pget.php on line 534       J+Clin+Microbiol 2019 ; 58 (1): ä Nephropedia Template TP {{tp|p=31666364|t=2019. Metagenomic Nanopore Sequencing of Influenza Virus Direct from Clinical Respiratory Samples |pdf=|usr=}}{{31666364}} gab.com Text Metagenomic Nanopore Sequencing of Influenza Virus Direct from Clinical Respiratory Samples JO: J Clin Microbiol http://www.kidney.de/pget.php?&tw=1&pmid=31666364 #FOAvip Influenza is a major global public health threat as a result of its highly pathogenic variants, large zoonotic reservoir, and pandemic potential. Metagenomic viral sequencing offers the potential for a diagnostic test for influenza virus which also provides insights on transmission, evolution, and drug resistance and simultaneously detects other viruses. We therefore set out to apply the Oxford Nanopore Technologies sequencing method to metagenomic sequencing of respiratory samples. We generated influenza virus reads down to a limit of detection of 10(2) to 10(3) genome copies/ml in pooled samples, observing a strong relationship between the viral titer and the proportion of influenza virus reads (P = 4.7 x 10(-5)). Applying our methods to clinical throat swabs, we generated influenza virus reads for 27/27 samples with mid-to-high viral titers (cycle threshold [C(T) ] values, <30) and 6/13 samples with low viral titers (C(T) values, 30 to 40). No false-positive reads were generated from 10 influenza virus-negative samples. Thus, Nanopore sequencing operated with 83% sensitivity (95% confidence interval [CI], 67 to 93%) and 100% specificity (95% CI, 69 to 100%) compared to the current diagnostic standard. Coverage of full-length virus was dependent on sample composition, being negatively influenced by increased host and bacterial reads. However, at high influenza virus titers, we were able to reconstruct >99% complete sequences for all eight gene segments. We also detected a human coronavirus coinfection in one clinical sample. While further optimization is required to improve sensitivity, this approach shows promise for the Nanopore platform to be used in the diagnosis and genetic analysis of influenza virus and other respiratory viruses. Twit Text FOAVip Metagenomic Nanopore Sequencing of Influenza Virus Direct from Clinical Respiratory Samples ????J Clin Microbiol http://www.kidney.de/pget.php?&tw=1&pmid=31666364 #FOAvip Twit Text # Free Paper on # # # # # LINK http://www.kidney.de/pget.php?&tw=1&pmid=31666364 #FOAvip English Wikipedia <ref>{{Cite pmid|31666364}}</ref> Metagenomic Nanopore Sequencing of Influenza Virus Direct from Clinical Respiratory Samples #MMPMID31666364 Lewandowski K; Xu Y; Pullan ST; Lumley SF; Foster D; Sanderson N; Vaughan A; Morgan M; Bright N; Kavanagh J; Vipond R; Carroll M; Marriott AC; Gooch KE; Andersson M; Jeffery K; Peto TEA; Crook DW; Walker AS; Matthews PC J Clin Microbiol 2019[Dec]; 58 (1): ä PMID31666364show ga Influenza is a major global public health threat as a result of its highly pathogenic variants, large zoonotic reservoir, and pandemic potential. Metagenomic viral sequencing offers the potential for a diagnostic test for influenza virus which also provides insights on transmission, evolution, and drug resistance and simultaneously detects other viruses. We therefore set out to apply the Oxford Nanopore Technologies sequencing method to metagenomic sequencing of respiratory samples. We generated influenza virus reads down to a limit of detection of 10(2) to 10(3) genome copies/ml in pooled samples, observing a strong relationship between the viral titer and the proportion of influenza virus reads (P = 4.7 x 10(-5)). Applying our methods to clinical throat swabs, we generated influenza virus reads for 27/27 samples with mid-to-high viral titers (cycle threshold [C(T) ] values, <30) and 6/13 samples with low viral titers (C(T) values, 30 to 40). No false-positive reads were generated from 10 influenza virus-negative samples. Thus, Nanopore sequencing operated with 83% sensitivity (95% confidence interval [CI], 67 to 93%) and 100% specificity (95% CI, 69 to 100%) compared to the current diagnostic standard. Coverage of full-length virus was dependent on sample composition, being negatively influenced by increased host and bacterial reads. However, at high influenza virus titers, we were able to reconstruct >99% complete sequences for all eight gene segments. We also detected a human coronavirus coinfection in one clinical sample. While further optimization is required to improve sensitivity, this approach shows promise for the Nanopore platform to be used in the diagnosis and genetic analysis of influenza virus and other respiratory viruses. |*Metagenomics/methods[MESH] |*Nanopore Sequencing/methods[MESH] |Computational Biology/methods[MESH] |England/epidemiology[MESH] |Genome, Viral[MESH] |High-Throughput Nucleotide Sequencing[MESH] |Humans[MESH] |Influenza, Human/diagnosis/epidemiology/*virology[MESH] |Orthomyxoviridae/classification/*genetics[MESH] |Phylogeny[MESH] |RNA Viruses/classification/genetics[MESH]Metagenomic Nanopore Sequencing of Influenza Virus Direct from Clinica(epmc).pdf DeepDyve Pubget Overpricing