External Contamination in Single Cell mtDNA Analysis 英文参考文献.docVIP

ExternalContaminationinSingleCellmtDNAAnalysis Yong-GangYao1*,Hans-Ju¨rgenBandelt2,NealS.Young1 1Hematology Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland, United States of America, 2DepartmentofMathematics,UniversityofHamburg,Hamburg,Germany Background. Mitochondrial DNA (mtDNA) variation in single hematopoietic cells, muscle fibers, oocytes, and from tiny amount of tumor tissues and degraded clinical specimens has been reported in many medical publications. External DNA contamination, notoriously difficult to avoid, threatens the integrity of such studies. Methodology/Principal Findings. Employing a phylogenetic approach, we analyzed the geographic origins of mtDNA sequence anomalies observed during multiplestudiesofmtDNAsequencevariationinatotalof7094singlehematopoieticcells.40eventswithirregularmtDNA patternsweredetected:eightinstances(fromsevendifferenthaplotypes)couldbetracedtolaboratorypersonnel;sixcases werecausedbysamplecross-contamination.Thesourcesoftheremainingeventscouldnotbeidentified,andtheanomalous sequence variation referred to matrilines from East Asia, Africa, or West Eurasia, respectively. These mtDNA sequence anomalies could be best explained by contamination. Conclusions. Using the known world mtDNA phylogeny, we could distinguishthegeographicoriginoftheanomalousmtDNAtypes,providingsomeusefulinformationregardingthesourceof contamination.OurdatasuggestthatroutinemtDNAsequenceanalysisoflaboratorypersonnelisinsufficienttoidentifyand eliminateallcontaminants.Arateof0.6%ofexternalcontaminationinthisstudy,whilelow,isnotnegligible:Unrecognized contaminantswillbemistakenasevidenceofremarkablesomaticmutationsassociatedwiththedevelopmentofcancerand otherdiseases.Theeffectivecontaminationratecanincreasebyafactorofmorethananorderofmagnitudeinsomestudies that did not institute high standards. Our results are of particular relevance to mtDNA research in medicine, and such an approachshouldbeadoptedtomai