摘要
Objective:ThisstudyaimstoestablishamethodforhighlyparallelmultiplexeddetectionofgeneticmutationsinChineselungcancersamplesthroughAgenaiPLEXchemistryandmatrix-assistedlaserdesorptionionizationtime-of-flightanalysisonMassARRAYmassspectrometryplatform.Methods:Wereviewedtherelatedliteratureanddataonlungcancertreatments.Wealsoidentified99mutationhotspotsin13targetgenescloselyrelatedtothepathogenesis,drugresistance,andmetastasisoflungcancer.Atotalof297primers,composedof99pairedforwardandreverseamplificationprimersand99matchedextensionprimers,weredesignedusingAssayDesignsoftware.Thedetectionmethodwasestablishedbyanalyzingeightcelllinesandsixlungcancerspecimens.TheproposedmethodwasthenvalidatedthroughcomparisonsbyusingaLungCarta~(TM)kit.ThesensitivityandspecificityoftheproposedmethodwereevaluatedbydirectlysequencingEGFRandKRASgenesin100lungcancercases.Results:Theproposedmethodwasabletodetectmultiplexgeneticmutationsinlungcancercelllines.Thisfindingwasconsistentwiththeobservationsonpreviouslyreportedmutations.Theproposedmethodcanalsodetectsuchmutationsinclinicallungcancerspecimens.ThisresultwasconsistentwiththeobservationswithLungCarta~(TM)kit.However,anFGFR2mutationwasdetectedonlythroughtheproposedmethod.Themeasuredsensitivityandspecificitywere100%and96.3%,respectively.Conclusions:TheproposedMassARRAYtechnology-basedmultiplexmethodcandetectgeneticmutationsinChineselungcancerpatients.Therefore,theproposedmethodcanbeappliedtodetectmutationsinothercancertissues.
出版日期
2016年01月11日（中国期刊网平台首次上网日期，不代表论文的发表时间）
