应用基因芯片方法检测结核分枝杆菌利福平和异烟肼的耐.pdf

680中国防痨杂志2011年10月第33卷第10期ChinJAntituberc,October2011,Vol33,No10

论著

应用基因芯片方法检测结核分枝杆菌利福平
和异烟肼的耐药性

张俊仙吴雪琼阳幼荣梁艳

【摘要】目的应用基因芯片方法检测结核分枝杆菌(Mycobacteriumtuberculosis,Mtb)对利福平和异烟肼的
耐受性,评价其临床应用价值。方法应用聚合酶链反应(polymerasechainreaction,PCR)扩增-基因芯片杂交的
方法检测经常规药敏实验证实的30株Mtb利福平和异烟肼敏感株和50株耐利福平和异烟肼分离株的rpoB基因
及katG和inhA基因突变,同时以PCR-直接测序法为对照。结果应用PCR-基因芯片与基因测序方法检测30株
Mtb利福平敏感株rpoB基因和异烟肼敏感株katG基因和inhA基因均为野生型。50株Mtb利福平耐药株中,
PCR-基因芯片与基因测序分析3株rpoB基因均为野生型,41株均为突变型;6株PCR-基因芯片与基因测序结果
不一致。50株Mtb异烟肼耐药株中,PCR-基因芯片与基因测序分析16株katG基因和30株inhA基因均为野生
型,31株katG基因均为315位密码子突变,7株inhA基因均为-15位突变型,其中2株为katG和inhA双重突变;
3株katG和13株inhAPCR-基因芯片与基因测序结果不一致。结论应用PCR-基因芯片方法可快速、有效地检
出大多数Mtb耐多药分离株,指导临床用药。
【关键词】分枝杆菌,结核;利福平;异烟肼;抗药性,细菌;寡核苷酸序列分析

RapiddetectionofMycobacteriumtuberculosisresistancetorifampicinandisoniazidbygenechipZHANGJun-
xian,WUXue-qiong,YANGYou-rong,LIANGYan.ArmyTuberculosisPreventionandControlKeyLaboratory,
InstituteforTuberculosisResearch,The309thHospitalofChinesePeopleLiberationArmy,Beijing100091,China
Correspondingauthor:WUXue-qiong,Email:[e-mail]
【Abstract】ObjectiveTodetectMycobacteriumtuberculosis(Mtb)resistancetorifampicin(RIF)andisoniazid
(INH)bygenechip,andtoevaluatetheirclinicalvalues.MethodsThirtyRIF-andINH-susceptibleand50RIF-
andINH-resistantclinicalisolatesinMtbwereanalyzedthemutationsofrpoB、katGandinhAgenesby(PCR)-gene
chipandDNAsequencing.ResultsTherpoB、katGandinhAgenesfrom30RIF-andINH-susceptibleisolatesof
MtbwereallwildtypesbyPCR-genechipandDNAsequencing.Of50RIF-resistantclinicalisolates,3hadwild
typesofropBgenes,and41hadropBmutationsbyPCR-genechipandDNAsequencing.Sixhadinconsistentre-
sultsbetweenPCR-genechipandDNAsequencing.Of50INH-resistantclinicalisolates,16hadwildtypesofkatG
genes,and31hadkatGmutationsoncodon315;30hadwildtypesofinhAgenes,and7hadinhAmutationsatpo-
sition-15.KatGresultsof3isolatesandinhAresultsof13isolateswereinconsistentbetweenPCR-genechipand
DNAsequencing.ConclusionPCR-genechipmightbearapidandeffectivemethodforthedetectionofMtb
mult-idrug-