Matches in SemOpenAlex for { <https://semopenalex.org/work/W2024215074> ?p ?o ?g. }
- W2024215074 endingPage "e0121948" @default.
- W2024215074 startingPage "e0121948" @default.
- W2024215074 abstract "There are increasing reports of plasma miRNAs as biomarkers of human disease but few standards in methodologic reporting, leading to inconsistent data. We systematically reviewed plasma miRNA studies published between July 2013-June 2014 to assess methodology. Six parameters were investigated: time to plasma extraction, methods of RNA extraction, type of miRNA, quantification, cycle threshold (Ct) setting, and methods of statistical analysis. We compared these data with a proposed standard methodologic technique. Beginning with initial screening for 380 miRNAs using microfluidic array technology and validation in an additional cohort of patients, we compared 11 miRNAs that exhibited differential expression between 16 patients with benign colorectal neoplasms (advanced adenomas) and 16 patients without any neoplasm (controls). Plasma was isolated immediately, 12, 24, 48, or 72 h following phlebotomy. miRNA was extracted using two different techniques (Trizol LS with pre-amplification or modified miRNeasy). We performed Taqman-based RT-PCR assays for the 11 miRNAs with subsequent analyses using a variable Ct setting or a fixed Ct set at 0.01, 0.03, 0.05, or 0.5. Assays were performed in duplicate by two different operators. RNU6 was the internal reference. Systematic review yielded 74 manuscripts meeting inclusion criteria. One manuscript (1.4%) documented all 6 methodological parameters, while < 5% of studies listed Ct setting. In our proposed standard technique, plasma extraction ≤12 h provided consistent ΔCt. miRNeasy extraction yielded higher miRNA concentrations and fewer non-expressed miRNAs compared to Trizol LS (1/704 miRNAs [0.14%] vs 109/704 miRNAs [15%], not expressed, respectively). A fixed Ct bar setting of 0.03 yielded the most reproducible data, provided that <10% miRNA were non-expressed. There was no significant intra-operator variability. There was significant inter-operator variation using Trizol LS extraction, while this was negligible using modified miRNeasy. For standardized reporting, we recommend plasma extraction ≤ 12 h, using modified miRNeasy extraction and utilizing a 0.03 Ct." @default.
- W2024215074 created "2016-06-24" @default.
- W2024215074 creator A5009507765 @default.
- W2024215074 creator A5011769829 @default.
- W2024215074 creator A5014771226 @default.
- W2024215074 creator A5043394178 @default.
- W2024215074 creator A5046209577 @default.
- W2024215074 creator A5059971466 @default.
- W2024215074 creator A5063421502 @default.
- W2024215074 date "2015-04-08" @default.
- W2024215074 modified "2023-10-16" @default.
- W2024215074 title "Assay Reproducibility in Clinical Studies of Plasma miRNA" @default.
- W2024215074 cites W113731422 @default.
- W2024215074 cites W119101192 @default.
- W2024215074 cites W1515659971 @default.
- W2024215074 cites W1574981725 @default.
- W2024215074 cites W1588142016 @default.
- W2024215074 cites W1588752417 @default.
- W2024215074 cites W1647075334 @default.
- W2024215074 cites W1798700676 @default.
- W2024215074 cites W1908410290 @default.
- W2024215074 cites W191278184 @default.
- W2024215074 cites W1964416456 @default.
- W2024215074 cites W1964753012 @default.
- W2024215074 cites W1966357431 @default.
- W2024215074 cites W1968008181 @default.
- W2024215074 cites W1969450948 @default.
- W2024215074 cites W1971764287 @default.
- W2024215074 cites W1974540586 @default.
- W2024215074 cites W1975862909 @default.
- W2024215074 cites W1976036380 @default.
- W2024215074 cites W1980455524 @default.
- W2024215074 cites W1981777743 @default.
- W2024215074 cites W1983932066 @default.
- W2024215074 cites W1993850432 @default.
- W2024215074 cites W1996072437 @default.
- W2024215074 cites W1998039339 @default.
- W2024215074 cites W1998915895 @default.
- W2024215074 cites W2001895697 @default.
- W2024215074 cites W2002636093 @default.
- W2024215074 cites W2003334487 @default.
- W2024215074 cites W2004153018 @default.
- W2024215074 cites W2008014951 @default.
- W2024215074 cites W2011960295 @default.
- W2024215074 cites W2014007818 @default.
- W2024215074 cites W2022018600 @default.
- W2024215074 cites W2022120322 @default.
- W2024215074 cites W2023642368 @default.
- W2024215074 cites W2032801250 @default.
- W2024215074 cites W2033121192 @default.
- W2024215074 cites W2033551842 @default.
- W2024215074 cites W2033892993 @default.
- W2024215074 cites W2037631027 @default.
- W2024215074 cites W2037764834 @default.
- W2024215074 cites W2038371585 @default.
- W2024215074 cites W2039232562 @default.
- W2024215074 cites W2042317364 @default.
- W2024215074 cites W2043469561 @default.
- W2024215074 cites W2043712800 @default.
- W2024215074 cites W2048063575 @default.
- W2024215074 cites W2048752013 @default.
- W2024215074 cites W2055471397 @default.
- W2024215074 cites W2057082801 @default.
- W2024215074 cites W2057588450 @default.
- W2024215074 cites W2062690993 @default.
- W2024215074 cites W2063099809 @default.
- W2024215074 cites W2064095494 @default.
- W2024215074 cites W2066740876 @default.
- W2024215074 cites W2067680105 @default.
- W2024215074 cites W2070425448 @default.
- W2024215074 cites W2073423022 @default.
- W2024215074 cites W2074311819 @default.
- W2024215074 cites W2074689365 @default.
- W2024215074 cites W2077173951 @default.
- W2024215074 cites W2078315764 @default.
- W2024215074 cites W2081064968 @default.
- W2024215074 cites W2086276344 @default.
- W2024215074 cites W2087876911 @default.
- W2024215074 cites W2093330545 @default.
- W2024215074 cites W2103950317 @default.
- W2024215074 cites W2105726520 @default.
- W2024215074 cites W2105826816 @default.
- W2024215074 cites W2111100167 @default.
- W2024215074 cites W2112582093 @default.
- W2024215074 cites W2113374205 @default.
- W2024215074 cites W2115093090 @default.
- W2024215074 cites W2115614933 @default.
- W2024215074 cites W2116069971 @default.
- W2024215074 cites W2122321093 @default.
- W2024215074 cites W2129570960 @default.
- W2024215074 cites W2130819396 @default.
- W2024215074 cites W2131130694 @default.
- W2024215074 cites W2141725382 @default.
- W2024215074 cites W2147493587 @default.
- W2024215074 cites W2147579615 @default.
- W2024215074 cites W2149423226 @default.
- W2024215074 cites W2150027485 @default.
- W2024215074 cites W2152070471 @default.