国际检验医学杂志
國際檢驗醫學雜誌
국제검험의학잡지
INTERNATIONAL JOURNAL OF LABORATORY MEDICINE
2008年
8期
679-682
,共4页
朱中元%王海波%谢勇%谢萌%王莉%朱义明%郭洁
硃中元%王海波%謝勇%謝萌%王莉%硃義明%郭潔
주중원%왕해파%사용%사맹%왕리%주의명%곽길
结核分支杆菌%利福平%rpoβ基因芯片
結覈分支桿菌%利福平%rpoβ基因芯片
결핵분지간균%리복평%rpoβ기인심편
Mycobacterium tuberculosis%rpoβ%Microarray
目的 建立基因芯片检测结核分支杆菌耐利福平基因突变,结合基因测序和常规药物敏感性试验进行比较,探讨其实用价值.方法 根据结核菌rpoβ基因突变特点.设计不同的寡核苷酸探针,点样制备可检测rpoβ基因突变的芯片,检测利福平(Rifompicin,RFP)株的rpoβ变异情况.结果 137株临床分离结核菌,49株耐RFP,占35.8%,平均MIC为179.6±135.0 μg/mL.Rpoβ突变率为95.7%(47/49),主要为点突变.突变点依次为531 Ser(TCG)→Leu(TTG)(40.8%)、531 Ser (TCG)→Trp(TGG)(12.2%)、526His(CAC)→Tyr(TAC)(6.1%)、526His(CAC)→Asp(GAC)(6.1%)、526His(CAC)→Asn(AAC)(6.1%)、526His(CAC)→Pro(CCC)(8.2%)、516Asp(GAC)→Val(GTC)(12.2%)、533Leu(CTG)→Arg(CGG)(4.1%)和513GIn(CAA)→Pro(CCA)(2.1%).结论 本研究建立的检测结核菌rpoβ基因突变的基因芯片技术敏感性高,特异性强,可应用于结核菌耐RFP基因的检测.
目的 建立基因芯片檢測結覈分支桿菌耐利福平基因突變,結閤基因測序和常規藥物敏感性試驗進行比較,探討其實用價值.方法 根據結覈菌rpoβ基因突變特點.設計不同的寡覈苷痠探針,點樣製備可檢測rpoβ基因突變的芯片,檢測利福平(Rifompicin,RFP)株的rpoβ變異情況.結果 137株臨床分離結覈菌,49株耐RFP,佔35.8%,平均MIC為179.6±135.0 μg/mL.Rpoβ突變率為95.7%(47/49),主要為點突變.突變點依次為531 Ser(TCG)→Leu(TTG)(40.8%)、531 Ser (TCG)→Trp(TGG)(12.2%)、526His(CAC)→Tyr(TAC)(6.1%)、526His(CAC)→Asp(GAC)(6.1%)、526His(CAC)→Asn(AAC)(6.1%)、526His(CAC)→Pro(CCC)(8.2%)、516Asp(GAC)→Val(GTC)(12.2%)、533Leu(CTG)→Arg(CGG)(4.1%)和513GIn(CAA)→Pro(CCA)(2.1%).結論 本研究建立的檢測結覈菌rpoβ基因突變的基因芯片技術敏感性高,特異性彊,可應用于結覈菌耐RFP基因的檢測.
목적 건립기인심편검측결핵분지간균내리복평기인돌변,결합기인측서화상규약물민감성시험진행비교,탐토기실용개치.방법 근거결핵균rpoβ기인돌변특점.설계불동적과핵감산탐침,점양제비가검측rpoβ기인돌변적심편,검측리복평(Rifompicin,RFP)주적rpoβ변이정황.결과 137주림상분리결핵균,49주내RFP,점35.8%,평균MIC위179.6±135.0 μg/mL.Rpoβ돌변솔위95.7%(47/49),주요위점돌변.돌변점의차위531 Ser(TCG)→Leu(TTG)(40.8%)、531 Ser (TCG)→Trp(TGG)(12.2%)、526His(CAC)→Tyr(TAC)(6.1%)、526His(CAC)→Asp(GAC)(6.1%)、526His(CAC)→Asn(AAC)(6.1%)、526His(CAC)→Pro(CCC)(8.2%)、516Asp(GAC)→Val(GTC)(12.2%)、533Leu(CTG)→Arg(CGG)(4.1%)화513GIn(CAA)→Pro(CCA)(2.1%).결론 본연구건립적검측결핵균rpoβ기인돌변적기인심편기술민감성고,특이성강,가응용우결핵균내RFP기인적검측.
Objective To establish the DNA probe microarray for detection M.tuberculosis rpoβ gene mutations associated with rifampicin(RFP)resistance,and evaluate its applicable value.Methods According to the characteristic of M tuberculosis rpoβ gene mutation related to RFP resistante.oligonucleotide probes were designed and spotted to prepare DNA chip.The detection efficacy was compared with DNA direct sequencing of the rpoβ and drug susceptibility test.Results 49 out of 1 37(35.8%)M.tuberculosis isolales were resistant to RFP with the average minimal inhibitory concentration of(179.6±135.0)μg/mL.The mutation rate of rpoβwas 95.7%(47/49),mainly point mutation.The mutationaI sites in order were 531Ser(TCG)→Leu(TTG)(40.8%),531 Set(TCG)→Trp(TGG)(12.2%),526His(CAC)→Tyr(TAC)(6.1%),526His(CAC)→Asp(GAC)(6.1%),526His(CAC)→Asn(AAC)(6.1%),526His(CAC)→Pro(CCC)(8.2%).516Asp(GAC)→Val(GTC)(12.2%),533Leu(CTG)→Arg(CGG)(4.1%),and 513GIn(CAA)→Pro(CCA)(2.1%).Conclusion The established DNA probe microarray is specific and sensitive for determination of M.tuberculosis resistant to RFP,and may be an effective tool for rapid determination of M.tuberculosis resistant to RFP in clinicalIaboratories.
