{"title":"How to make Mathematics Biology's next and better microscope","authors":"Jim Huggett, Justin O’Grady, Stephen Bustin","doi":"10.1016/j.bdq.2014.09.001","DOIUrl":"10.1016/j.bdq.2014.09.001","url":null,"abstract":"","PeriodicalId":38073,"journal":{"name":"Biomolecular Detection and Quantification","volume":"1 1","pages":"Pages A1-A3"},"PeriodicalIF":0.0,"publicationDate":"2014-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.bdq.2014.09.001","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"54133470","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Coren A. Milbury , Qun Zhong , Jesse Lin, Miguel Williams, Jeff Olson, Darren R. Link, Brian Hutchison
{"title":"Determining lower limits of detection of digital PCR assays for cancer-related gene mutations","authors":"Coren A. Milbury , Qun Zhong , Jesse Lin, Miguel Williams, Jeff Olson, Darren R. Link, Brian Hutchison","doi":"10.1016/j.bdq.2014.08.001","DOIUrl":"10.1016/j.bdq.2014.08.001","url":null,"abstract":"<div><p>Digital PCR offers very high sensitivity compared to many other technologies for processing molecular detection assays. Herein, a process is outlined for determining the lower limit of detection (LoD) of two droplet-based digital PCR assays for point mutations of the epidermal growth factor receptor (<em>EGFR</em>) gene. Hydrolysis probe mutation-detection assays for <em>EGFR</em> p.L858R and p.T790M mutations were characterized in detail. Furthermore, sixteen additional cancer-related mutation assays were explored by the same approach. For the <em>EGFR</em> L8585R assay, the assay sensitivity is extremely good, and thus, the LoD is limited by the amount of amplifiable DNA that is analyzed. With 95% confidence limits, the LoD is one mutant in 180,000 wild-type molecules for the evaluation of 3.3<!--> <!-->μg of genomic DNA, and detection of one mutant molecule in over 4 million wild-type molecules was achieved when 70 million copies of DNA were processed. The measured false-positive rate for the <em>EGFR</em> L8585R assay is one in 14 million, which indicates the theoretical LoD if an unlimited amount of DNA is evaluated. For the <em>EFGR</em> T790M assay, the LoD is one mutant in 13,000 for analysis of a 3.3<!--> <!-->μg sample of genomic DNA, and the dPCR assay limit sensitivity approaches one mutant in 22,000 wild-type molecules.</p></div>","PeriodicalId":38073,"journal":{"name":"Biomolecular Detection and Quantification","volume":"1 1","pages":"Pages 8-22"},"PeriodicalIF":0.0,"publicationDate":"2014-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.bdq.2014.08.001","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"54133400","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}