不同粪便DNA提取试剂盒及样本储_省略_对粪便人基因组DNA提取率的影响_王勇

53-56．10deVriesACvanGriekenNCLoomanCWetal．GastriccancerriskinpatientswithpremalignantgastriclesionsanationwidecohortstudyintheNetherlandsJ．Gastroenterology20081344945-952．11．J．201225765-68．12JacksonPGrimmMOKingsleyEAetal．ＲelationshipbetweenexpressionofKAI1metastasissuppressorgenemＲNAlevelsandp53inhumanbladderandprostatecancercelllinesJ．UrolOncol20027399-104．13．P53J．201027160-65．14YamadaHShinmuraKOkudelaKetal．Identifica-tionandcharacterizationofanovelgermlinep53muta-tioninfamilialgastriccancerintheJapanesepopulationJ．Carcinogenesis20072892013-2018．15．J．200831229-11．16BritoMJWilliamsGTThompsonHetal．Expressionofp53inearlyT1gastriccarcinomaandprecancerousadjacentmucosaJ．Gut199435121697-1700．17．P53J．201015227．18．p53J．20083441-37．19．P53CerbB-2J．200614302943-2947．20MaYLPengJYZhangPetal．Immunohistochemi-calanalysisrevealedCD34andKi67proteinexpressionassignificantprognosticfactorsincolorectalcancerJ．MedOncol2010272304-309．21．104J．2009382100-105．2013-12-172014-02-11DNADNA2013ZX1000410321275161－BL2012061210009、、、210002、、、、210002、、、、[e-mail]。18QIAampDNAStoolMiniKitMOBIOUltraCleanFecalDNAIsolationKitE．Z．N．A．MagBindStoolDNAKit3DNADNADNAPCＲDNA。3QIAampDNAStoolMiniKitDNA67．81ng/gE．Z．N．A．MagBindStoolDNAKitA260/A2802．07MOBIOUltraCleanFecalDNAIsolationKit40min－70℃30dDNAP＜0.01。QIAampDNAStoolMiniKitDNA－70℃30dDNA。DNAＲ446A1002-3429201404-0077-04DOI10．3969/j．issn．1002-3429．2014．04．033InfluenceofDifferentStorageTimesandDNAzolＲeagentKitsontheEfficiencyofHumanGenomicDNAExtractionfromStools·77·20144274ClinicalMisdiagnosis＆MistherapyVol．27No．4April2014WANGYong123ZOUBing-jie23LIUYun-long123LISi-qin123ZHOUGuo-hua1231．SchoolofLifeScienceandTech-nologyChinaPharmaceuticalUniversityNanjing210009China2．DepartmentofPharmacologyClinicalCollegeofMedicalSchoolofNanjingUniversityNanjingGeneralHospitalofNanjingMilitaryAreaCommandNanjing210002China3．HuadongＲesearchInstituteforMedicineandBiotechnologiesNanjing210002ChinaAbstractObjectiveTocomparetheinfluenceofdifferentDNAzolreagentkitsandstoragetimesontheefficiencyofhu-mangenomicDNAextractionfromstools．MethodsDNAwasextractedfromstoolsof18healthypeoplefollowingmedicalexamina-tionsbyusingthreedifferentstoolDNAextractionkitsQIAampDNAStoolMiniKitMOBIOUltraCleanFecalDNAIsolationKitandE．Z．N．A．MagBindStoolDNAKit．ThequantityandpurityofthetotalDNAwereanalyzedwithultravioletspectrophotome-tryandthequantitationoftheadenomatouspolyposiscoliiAPCgeneinthetotalDNAwasdetectedwithreal-timepolymerasechainreactionPCＲforcomparingthedifferenceofhumangenomicDNAextractingefficiencyandtheinfluencebydifferentstor-agetimes．ＲesultsAmongthethreedifferentkitsQIAGENDNAStoolMinikitshowedthehighestyieldofhumangenomicDNAaveraged67．81ng/gtheE．Z．N．A．MagBindStoolDNAkithadthehighestpurityA260/A280averaged2．07MOBIOUl-traCleanFecalDNAIsolationkithadtheshortestoperativetimesuckabout40minforasample．Thestoolsampleswerestoredat－70℃within30dthedifferencesofefficiencyoftotalDNAextractionwerestatisticallysignificantP＜0.01whilethediffer-encesofefficiencyofhumangenomicDNAextractionwerenotstatisticallysignificantintermsofdifferentstoragetimesP＞0．05．ConclusionQIAGENkithasthehighestefficiencyforhumangenomicDNAextractionfromstoolsandstoolsamplesstoreat－70℃within30dshownoinfluenceontheextractionefficiencyofhumangenomicDNA．KeywordsＲeagentkitdiagnosticStoragetimeDNAFecesDNA1-3DNA。、DNADNADNA4-8。DNA9-13DNADNA。DNADNADNA。3DNADNA－70℃30dDNA。11.11820128—9144。2h－70℃。1.2QIAampDNAStoolMiniKitQIAGENMOBIOUltraCleanFecalDNAIsolationKitMOBIOE．Z．N．A．MagBindStoolDNAKitOMEGATakaraPremixExTaqOneDropOD-1000SpectrophotometerStepOneＲeal-TimePCＲSystemABI。1.3ad-enomatouspolyposiscoliAPCTaqManPrimerPremier5．0TaqMan5'-FAM-CCCTGGACAAACCATGCCACCAA-ECLIPSE-3'5'-AGCCCCAGTGATCTTCCAGAT-3'5'-AGGTGGTGGAGGTGTTTTACTTCT-3'72bp。1.4DNA3DNA18DNADNA－20℃。DNAOneDropOD-1000260nm280nmPCＲDNAAPCDNAPCＲ25μlDNA5μl5μl2×PremixExTaqProbeqPCＲ12．5μl10μmol/L0．5μl10μmol/L0．5μl10μmol/LTaqMan0．5μl50×ＲoxＲefer-enceDye0．5μl5．5μl。95℃30s95℃5s60℃31s40。DNA20、100、500、2500、12500/5μl514Ｒ2＞0．99。1.517·87·20144274ClinicalMisdiagnosis＆MistherapyVol．27No．4April20143317DNADNAA260/A280DNAPCＲAPC6-714-15DNA。3、6。DNA210．20g－70℃0、1、3、6、10、2030dQIAampDNAStoolMiniKit3DNADNAPCＲAPCDNA。1.6IBMSPSSStatistics19One-wayANOVALeastSignificantDifferenceTamhane＇sT2。α=0．05。22.13DNAQIAampDNAStoolMiniKitDNADNA9、5DNA6、4P＜0．01MOBIOUltra-CleanFecalDNAIsolationKitDNAE．Z．N．A．MagBindStoolDNAKitP＜0．01。1。QIAampDNAStoolMiniKit1846/60minMOBIOUltraCleanFecalDNAIsolationKit2051/40minE．Z．N．A．MagBindStoolDNAKit2527/70min。2.2DNADNAP＜0.0101d、3～10dDNAP＞0.053～10d0～1dDNAP＜0.0120～30d3～10dDNAP＜0.05。1。DNAP＞0.05DNA0～30d39．66±5.62ng/g。2。13DNADNADNAμg/gDNAng/gDNADNA%DNAA260/A280QIAampDNAStoolMiniKit1789．42b67．81d7．56×10－22．51fE．Z．N．A．MagBindStoolDNAKit179．5911．8411．25×10－22．07MOBIOUltraCleanFecalDNAIsolationKit1717．52b17．858．65×10－22．08DNA3．3pg6E．Z．N．A．MagBindStoolDNAKitbP＜0.01dP＜0.01fP＜0.01d样本储存时间（d）总DNA提取量（μg/g）0%%%%%%%%%%%1%%%%%%%%%%3%%%%%%%%%%6%%%%%%%%%10%%%%%%%20%%%%%%%%30403020100aadd1DNA3～10daP＜0.050～1ddP＜0.01样本储存时间（d）人基因组DNA提取量（ng/g）0%%%