@article{Silva_Salta_Henrique_Jerónimo_2020, title={Comparison and Optimization of DNA Extraction Methods from Cervical Cells Collected in ThinPrep® PreservCyt}, url={https://parc.ipp.pt/index.php/citotech/article/view/3778}, DOI={10.26537/citotech.vi5.3778}, abstractNote={<p><span id="page3R_mcid44" class="markedContent"><span dir="ltr" style="left: 185.633px; top: 695.504px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.01605);" role="presentation">Cervical cancer is the ninth leading cancer in women in Portugal. Cervical cancer screening </span><br role="presentation" /><span dir="ltr" style="left: 150.04px; top: 721.337px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.01876);" role="presentation">methods have changed since the implementation of the cytology</span><span dir="ltr" style="left: 605.917px; top: 721.337px; font-size: 15px; font-family: sans-serif;" role="presentation">-</span><span dir="ltr" style="left: 610.917px; top: 721.337px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.02504);" role="presentation">based test developed by George </span><br role="presentation" /><span dir="ltr" style="left: 150.04px; top: 747.337px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.03533);" role="presentation">Papanicolaou </span><span dir="ltr" style="left: 251.71px; top: 747.337px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.0575);" role="presentation">until </span><span dir="ltr" style="left: 290.05px; top: 747.337px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.04);" role="presentation">the </span><span dir="ltr" style="left: 321.625px; top: 747.337px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.01908);" role="presentation">human </span><span dir="ltr" style="left: 378.175px; top: 747.337px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.03);" role="presentation">papillomavirus</span><span dir="ltr" style="left: 474.483px; top: 747.337px; font-size: 15px; font-family: sans-serif;" role="presentation">-</span><span dir="ltr" style="left: 479.483px; top: 747.337px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.02102);" role="presentation">based </span><span dir="ltr" style="left: 531.023px; top: 747.337px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.0175);" role="presentation">screening. </span><span dir="ltr" style="left: 610.928px; top: 747.337px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.03007);" role="presentation">Nowadays, </span><span dir="ltr" style="left: 696.117px; top: 747.337px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.01758);" role="presentation">new </span><span dir="ltr" style="left: 734.277px; top: 747.337px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.01942);" role="presentation">biomarkers </span><span dir="ltr" style="left: 819.387px; top: 747.337px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.0542);" role="presentation">like </span><br role="presentation" /><span dir="ltr" style="left: 150.04px; top: 773.137px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.00877);" role="presentation">deoxyribonucleic acid (DNA) methylation has been proposed as potential triage method of samples </span><br role="presentation" /><span dir="ltr" style="left: 150.04px; top: 798.937px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.02944);" role="presentation">testing positive for human papillomavirus </span> </span><span id="page3R_mcid45" class="markedContent"><br role="presentation" /><span dir="ltr" style="left: 185.633px; top: 824.937px; font-size: 15px; font-family: sans-serif; transform: scaleX(0.982735);" role="presentation">The aim of this study was to optimize and compare three different DNA extraction methods for </span><br role="presentation" /><span dir="ltr" style="left: 150.04px; top: 850.737px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.02825);" role="presentation">further methylation analysis. DNA from sixteen cervical samples collected in ThinPrep® PreservCyt was </span><br role="presentation" /><span dir="ltr" style="left: 150.04px; top: 876.537px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.01326);" role="presentation">extracted by the phenol</span><span dir="ltr" style="left: 310.483px; top: 876.537px; font-size: 15px; font-family: sans-serif;" role="presentation">-</span><span dir="ltr" style="left: 315.283px; top: 876.537px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.03019);" role="presentation">chloroform method, </span> <span dir="ltr" style="left: 451.683px; top: 876.537px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.05);" role="presentation">by </span> <span dir="ltr" style="left: 473.083px; top: 876.537px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.056);" role="presentation">QIAamp® MinElut</span><span dir="ltr" style="left: 594.717px; top: 876.537px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.08136);" role="presentation">e™ Media kit (QIAGEN, Hilden) and </span><br role="presentation" /><span dir="ltr" style="left: 150.04px; top: 902.537px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.05);" role="presentation">by </span> <span dir="ltr" style="left: 172.433px; top: 902.537px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.02171);" role="presentation">Blood DNA </span> <span dir="ltr" style="left: 255.633px; top: 902.537px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.00605);" role="presentation">Isolation Mini Kit (Norgen Biotek, Ontario). After quantification, DNA was modified by </span><br role="presentation" /><span dir="ltr" style="left: 150.04px; top: 928.337px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.06039);" role="presentation">Sodium Bisulfite and amplified by Quantitative Specific Methylation Polymerase Chain Reaction for β</span><span dir="ltr" style="left: 836.967px; top: 928.337px; font-size: 15px; font-family: sans-serif;" role="presentation">-</span><br role="presentation" /><span dir="ltr" style="left: 150.04px; top: 954.337px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.04944);" role="presentation">Actin gene. Finally, C</span><span dir="ltr" style="left: 291.833px; top: 954.337px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.02797);" role="presentation">ycle threshold (Ct) mean values were evaluated. </span> </span><span id="page3R_mcid46" class="markedContent"><br role="presentation" /><span dir="ltr" style="left: 185.633px; top: 980.137px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.04479);" role="presentation">The phenol</span><span dir="ltr" style="left: 261.033px; top: 980.137px; font-size: 15px; font-family: sans-serif;" role="presentation">-</span><span dir="ltr" style="left: 265.833px; top: 980.137px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.01512);" role="presentation">chloroform method</span> <span dir="ltr" style="left: 395.083px; top: 980.137px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.0343);" role="presentation">had the higher sample input and, consequently</span> <span dir="ltr" style="left: 710.967px; top: 980.137px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.03696);" role="presentation">allowed for a higher </span><br role="presentation" /><span dir="ltr" style="left: 150.04px; top: 1005.94px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.04872);" role="presentation">DNA quantity. Fifteen mL </span> <span dir="ltr" style="left: 339.083px; top: 1005.94px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.03387);" role="presentation">of sample allowed for get 13500</span><span dir="ltr" style="left: 573.117px; top: 1005.94px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.03952);" role="presentation">ng with the pheno</span><span dir="ltr" style="left: 704.317px; top: 1005.94px; font-size: 15px; font-family: sans-serif;" role="presentation">l</span><span dir="ltr" style="left: 707.717px; top: 1005.94px; font-size: 15px; font-family: sans-serif;" role="presentation">-</span><span dir="ltr" style="left: 712.567px; top: 1005.94px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.01714);" role="presentation">chloroform method </span><br role="presentation" /><span dir="ltr" style="left: 150.04px; top: 1031.99px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.03466);" role="presentation">meanwhile 2mL of s</span><span dir="ltr" style="left: 285.833px; top: 1031.99px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.04722);" role="presentation">ample provided 8940</span><span dir="ltr" style="left: 427.283px; top: 1031.99px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.0639);" role="presentation">ng with QIAa</span><span dir="ltr" style="left: 514.717px; top: 1031.99px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.05959);" role="presentation">mp® MinElute™ Media kit and 927</span><span dir="ltr" style="left: 750.167px; top: 1031.99px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.05115);" role="presentation">ng with Blood </span><br role="presentation" /><span dir="ltr" style="left: 150.04px; top: 1057.79px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.07027);" role="presentation">DNA Isolation Mini Kit. However</span><span dir="ltr" style="left: 366.483px; top: 1057.79px; font-size: 15px; font-family: sans-serif;" role="presentation">,</span> <span dir="ltr" style="left: 375.883px; top: 1057.79px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.01173);" role="presentation">commercial kits </span><span dir="ltr" style="left: 485.283px; top: 1057.79px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.04406);" role="presentation">presented higher efficiency and</span> <span dir="ltr" style="left: 701.717px; top: 1057.79px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.0438);" role="presentation">advantages as being </span><br role="presentation" /><span dir="ltr" style="left: 150.04px; top: 1083.59px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.02322);" role="presentation">less time consuming and easier to perform. No significant differences were obse</span><span dir="ltr" style="left: 683.917px; top: 1083.59px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.03051);" role="presentation">rved for amplification Ct </span><br role="presentation" /><span dir="ltr" style="left: 150.04px; top: 1109.59px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.03078);" role="presentation">values between the three DNA extraction methods neither between samples</span> <span dir="ltr" style="left: 661.117px; top: 1109.59px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.03677);" role="presentation">archived for ten or fourteen </span><br role="presentation" /><span dir="ltr" style="left: 150.04px; top: 1135.39px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.01184);" role="presentation">months. </span> </span><span id="page3R_mcid47" class="markedContent"><br role="presentation" /><span dir="ltr" style="left: 185.633px; top: 1161.19px; font-size: 15px; font-family: sans-serif; transform: scaleX(0.981406);" role="presentation">Concluding, the three methods provided DNA to amplification but the optimization of commercial </span><br role="presentation" /><span dir="ltr" style="left: 150.04px; top: 1187.19px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.03844);" role="presentation">kits turned out to be </span><span dir="ltr" style="left: 286.033px; top: 1187.19px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.02733);" role="presentation">more advantageous.</span></span></p>}, number={5}, journal={Citotech Online - Case Review}, author={Silva, Andreia and Salta, S and Henrique, R and Jerónimo, C}, year={2020}, month={Dec.}, pages={1–10} }