<?xml version="1.0" encoding="UTF-8"?><xml><records><record><source-app name="Biblio" version="7.x">Drupal-Biblio</source-app><ref-type>17</ref-type><contributors><authors><author><style face="normal" font="default" size="100%">Vorkas,P.A.</style></author><author><style face="normal" font="default" size="100%">Poumpouridou,N.</style></author><author><style face="normal" font="default" size="100%">Agelaki, S.</style></author><author><style face="normal" font="default" size="100%">Kroupis, C.</style></author><author><style face="normal" font="default" size="100%">Georgoulias, V.</style></author><author><style face="normal" font="default" size="100%">Lianidou,E.S.</style></author></authors></contributors><titles><title><style face="normal" font="default" size="100%">PIK3CA hotspot mutation scanning by a novel and highly sensitive high-resolution small amplicon melting analysis method</style></title><secondary-title><style face="normal" font="default" size="100%">J.Mol.Diagn.</style></secondary-title></titles><keywords><keyword><style  face="normal" font="default" size="100%">analysis</style></keyword><keyword><style  face="normal" font="default" size="100%">Base Sequence</style></keyword><keyword><style  face="normal" font="default" size="100%">Cell Line,Tumor</style></keyword><keyword><style  face="normal" font="default" size="100%">DNA Primers</style></keyword><keyword><style  face="normal" font="default" size="100%">genetics</style></keyword><keyword><style  face="normal" font="default" size="100%">Greece</style></keyword><keyword><style  face="normal" font="default" size="100%">Humans</style></keyword><keyword><style  face="normal" font="default" size="100%">Mutation</style></keyword><keyword><style  face="normal" font="default" size="100%">Paraffin Embedding</style></keyword><keyword><style  face="normal" font="default" size="100%">Phosphatidylinositol 3-Kinases</style></keyword><keyword><style  face="normal" font="default" size="100%">polymerase chain reaction</style></keyword><keyword><style  face="normal" font="default" size="100%">Reproducibility of Results</style></keyword><keyword><style  face="normal" font="default" size="100%">Sequence Analysis,DNA</style></keyword><keyword><style  face="normal" font="default" size="100%">therapy</style></keyword></keywords><dates><year><style  face="normal" font="default" size="100%">2010</style></year><pub-dates><date><style  face="normal" font="default" size="100%">2010/09//</style></date></pub-dates></dates><urls><web-urls><url><style face="normal" font="default" size="100%">PM:20616362</style></url></web-urls></urls><volume><style face="normal" font="default" size="100%">12</style></volume><pages><style face="normal" font="default" size="100%">697 - 704</style></pages><language><style face="normal" font="default" size="100%">eng</style></language><abstract><style face="normal" font="default" size="100%">Somatic mutations in the PIK3CA gene have been discovered in many human cancers, and their presence correlates to therapy response. Three &quot;hotspot&quot; mutations within the PIK3CA gene are localized in exons 9 and 20. High-resolution melting analysis (HRMA) is a highly sensitive, robust, rapid, and cost-effective mutation analysis technique. We developed a novel methodology for the detection of hotspot mutations in exons 9 and 20 of the PIK3CA gene that is based on a combination of PCR and HRMA. The PIK3CA HRMA assay was evaluated by performing repeatability, sensitivity, and comparison with DNA sequencing studies and was further validated in 129 formalin-fixed paraffin-embedded breast tissue samples: 99 tumors, 20 noncancerous, and 10 fibroadenomas. The developed methodology was further applied in a selected group of 75 breast cancer patients who underwent Trastuzumab treatment. In sensitivity studies, the assay presented a capability to detect as low as 1% of mutated dsDNA in the presence of wtDNA for both exons. In the 99 tumor samples (validation group), 12/99 (12.1%) exon 9 mutations and 20/99 (20.2%) exon 20 mutations were found. No mutations were found in noncancerous tissues. In fibroadenomas, we report one PIK3CA mutation for the first time. In the selected group, 30/75 (40%) samples were detected as mutants. The PIK3CA HRMA assay is highly sensitive, reliable, cost-effective, and easy-to-perform, and therefore can be used as a screening test in a high-throughput pharmacodiagnostic setting</style></abstract><issue><style face="normal" font="default" size="100%">5</style></issue><notes><style face="normal" font="default" size="100%">DA - 20100831IS - 1943-7811 (Electronic)IS - 1525-1578 (Linking)LA - engPT - Journal ArticlePT - Research Support, Non-U.S. Gov'tPT - Validation StudiesRN - 0 (DNA Primers)RN - EC 2.7.1.- (Phosphatidylinositol 3-Kinases)RN - EC 2.7.1.137 (PIK3CA protein, human)SB - IM</style></notes><custom5><style face="normal" font="default" size="100%">PMC2928435</style></custom5></record></records></xml>