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1. 第三军医大学西南医院检验科
2. 解放军第452医院医教部
纸质出版日期:2009,
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[1]陈庆海,府伟灵,薛强,张雪,华兴,匡红.发夹DNA探针检测乙胺丁醇耐药结核杆菌embB基因306密码子突变研究[J].重庆医科大学学报,2009,34(02):135-138.
CHEN Qin-hai, et al. The study on detecting mutation site of 306 codon in Ethambutol resistant embB gene of Mycobacterium tuberculosis bacterium by hair clamp probe[J]. Journal of Chongqing Medical University, 2009,34(2):135-138.
[1]陈庆海,府伟灵,薛强,张雪,华兴,匡红.发夹DNA探针检测乙胺丁醇耐药结核杆菌embB基因306密码子突变研究[J].重庆医科大学学报,2009,34(02):135-138. DOI:
CHEN Qin-hai, et al. The study on detecting mutation site of 306 codon in Ethambutol resistant embB gene of Mycobacterium tuberculosis bacterium by hair clamp probe[J]. Journal of Chongqing Medical University, 2009,34(2):135-138. DOI:
目的:针对结核杆菌耐乙胺丁醇embB306codon位点
设计发夹DNA探针及其相应的单侧延长臂发夹DNA探针
建立发夹DNA探针芯片技术
并初步运用荧光显微镜观测探针与扩增产物杂交的荧光信号。方法:运用Beacondesigner软件
设计embB基因包含306codon的发夹DNA探针及其相应的单侧延长臂发夹DNA探针
荧光显微镜检测embB306codon扩增片段与探针杂交后荧光信号
比较扩增产物测序结果。结果:通过荧光显微镜观测到结核标准株及embB306codon突变株PCR产物与探针杂交后荧光信号存在显著差异;33株耐乙胺丁醇组与10株H37RV标准株对照组荧光信号强度比较
耐乙胺丁醇组embB306codon突变检出率为66%
测序法突变检出率为69%。结论:embB306codon点突变是结核杆菌耐乙胺丁醇的主要原因;发夹DNA探针芯片技术可以有效检测单碱基靶点突变;应用荧光显微镜可以高灵敏地观测荧光芯片杂交靶点。
Objective:To design hair clamp probe and the probes with single extending arm detecting embB306codon of Ethambutol resis-tant Mycobacterium tuberculosis bacterium(MTB)
meanwhile
to design fair clamp probe chip and detecting fluorescence signal from hy-bridization between the amplified product and probe by fluorescence microscope.Methods:The software
Beacon designer
was used to de-sign fair clamp probe and the probes with single extending arm detecting embB303codon and detecting fluorescence signal from hybridiza-tion between the amplified product and probe
and confer to the sequencing results.Results:The difference between PCR products from standard strain and ethambutol resistant one was obvious in detecting the fluorescent light with fluorescence microscope.We detected fluo-rescent light signal between the 33 ethambutol resistant strains and 10 H37RV standard strains.The rate of resistant ethambutol detected with hair clamp probe was about 66%
and the rate of sequencing was about 69%.Conclusions:The mutation site of embB306codon of MTB is the main reason of ethambutol resistant MTB.The technology of fair clamp DNA probe chip can effectively detect mutation of single base site.Fluorescence microscope can sensitivily detect the site of hybridization on fluorescence chip.
耐乙胺丁醇embB基因306密码子点突变发夹DNA探针荧光显微镜
Ethambutol resistant embB gene306 Codon of mutation siteHair clamp probeFluorescence microscope
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