熒光微球技術(shù)在結(jié)核桿菌耐利福平菌株rpoB基因突變多重檢測(cè)中的應(yīng)用研究.pdf

1、西北大學(xué)碩士學(xué)位論文熒光微球技術(shù)在結(jié)核桿菌耐利福平菌株rpoB基因突變多重檢測(cè)中的應(yīng)用研究姓名：杜蓬申請(qǐng)學(xué)位級(jí)別：碩士專業(yè)：微生物學(xué)指導(dǎo)教師：王衛(wèi)衛(wèi)陳超2002.6.1墮!!查蘭堡主蘭垡堡莖——TheAppficationofFluorescentMicrosphereBasedTechnologyforMultiplexedDetectionofrpoBMutationsinRifampinResistantMycobactetium

2、tuberculosisIsolatesDuPengDirectedbyProfWangWeiweiandProfChenChao(BiochipR&DCenterSchoolofLifeScience，NofthwestUniversityChina)AbstractTheemergenceandspreadofdrugresistantMtubtr￡u／onsespeciallyrifampin—resistantMtuberⅢlo

3、sisisoneofimportantreasonsforincreasinglyseveretuberculosisIthadreported，mutationshavebeenfoundintherpoBgeneof96％ofrifampin—resistantclinicalMtubenMosisisolatesAconsiderableamountofmethodshavebeendevotedinthelastfewyears

4、tothedetectionofrpoBmutations，butmostofthemhavetheirowfllimitationsInthisstudy，afluorescentmicrosphere—basedtechnologyhadbeenintroducedtOthemultiplexeddetectionofrpoBmutationsinclinicalMtuberculosisisolatesAndthemainrese

5、archworkinthispaperwasdividedintothreeparts，(1)PCRAmplificationofrpoBhighvariationregion；(2)MultiplexeddetectionofrpoBmumfionsinclinicalrifampin—resistantMtuberculosisisolatesusingfivemicrosphereboundallelespecificprobes

6、；(3)Comparisonofmicrosphere—basedtechnology，rifampinsusceptibilitytestingDNAsequencingandPCR—SSCPanalysisfordetectionofrifampinresistanceinMtubenalosisItwasshowedintheresults，apointmutationineithercodon531or526wasobserve

7、din10of42clinicalMtubenwlosisisolatesandthisresultiSconcordantewi山phenotypicfiNmpinsusceptibilitytestingandDNAsequencingresultsThecomparisonofPCR—SSCPanalysisindicatedsomerifampin—resistantMtubaralodshadnotbeendetectedFh

8、eseresultssuggestedthatfluorescentmicrosphere—basedtechnologyisamoresensitiveandveracioustoolinmultiplexeddetectionofrpoBmutationsInaddition，byDNAsequencingdoublemutationsinasanlecodonwereobservedintwesistantstrains；more

9、thandoublemutationsindifierentcodonsofonefifampin—susceptiblestrainwerealSOdetected血ismaybetheeffectofsuppressormutation，KeyWords：dfampin—resistantMycobacteriumtubenulosisisolates；rpoBgene；fluorescentmicrosphere；muldplex