本文選題：結(jié)核分枝桿菌　切入點(diǎn)：耐多藥結(jié)核　出處：《湖南師范大學(xué)》2015年碩士論文

【摘要】：目的:調(diào)查結(jié)核患者中結(jié)核分枝桿菌的耐藥性,分析耐藥結(jié)核分枝桿菌臨床分離株的rpoB、gyrA和gyrB基因的突變情況,探討一線藥物利福平、二線藥物氧氟沙星的耐藥產(chǎn)生與rpoB基因、gyrA和gyrB基因突變的關(guān)系,為結(jié)核分枝桿菌(MTB)的耐藥性研究、耐多藥結(jié)核病(MDR-TB)和廣泛耐藥結(jié)核病(XDR-TB)的診斷與治療提供新的實(shí)驗(yàn)依據(jù)。方法:①收集湖南省胸科醫(yī)院6535例就診的肺結(jié)核患者的標(biāo)本,進(jìn)行結(jié)核分枝桿菌液體培養(yǎng)及絕對(duì)濃度法藥物敏感性實(shí)驗(yàn)(簡(jiǎn)稱“藥敏實(shí)驗(yàn)”,DST),檢測(cè)培養(yǎng)菌株對(duì)鏈霉素(SM,S)、異煙肼(INH,H)、利福平(RFP,R)、乙胺丁醇(EMB,E)、卡那霉素(KM,K)和氧氟沙星(OFX,O)6種藥物的敏感性,以結(jié)核分枝桿菌標(biāo)準(zhǔn)菌株H37Rv為對(duì)照,獲得菌株對(duì)6種藥物的耐藥譜。②選取利福平耐藥菌株,用超聲波裂解法提取MTB基因組DNA,對(duì)與利福平耐藥相關(guān)的rpoB基因進(jìn)行擴(kuò)增、測(cè)序并比對(duì)其突變情況,分析基因突變和耐藥表型的關(guān)系。③選取氧氟沙星耐藥菌株,采用超聲波裂解法提取MTB基因組DNA,對(duì)與氧氟沙星耐藥相關(guān)的gyrA和gyr B基因進(jìn)行擴(kuò)增、測(cè)序,分析該兩個(gè)基因突變與耐藥表型的相關(guān)性。結(jié)果:①結(jié)核分枝桿菌培養(yǎng)陽性1140例,初治患者826例,復(fù)治患者314例;耐藥肺結(jié)核患者443例,其中耐藥初治患者296例,耐藥復(fù)治患者147例,總耐藥率為38.85%(443/1140),初治患者總耐藥率為35.84%(296/826),復(fù)治患者總耐藥率為46.81%(147/314)。耐多藥患者270例,耐多藥率為23.68%(270/1140),其中同時(shí)耐利福平、異煙肼和鏈霉素在耐多藥模式中所占比例最高,達(dá)32.22%(87/270)。而單一耐藥以利福平最高,耐單藥率為28.51%(325/1140)。②本地區(qū)rpoB基因突變以C531T(Ser→Leu)為主,占69.64%(39/56),其中耐多藥菌株有29株(70.73%),其次是發(fā)生在RRDR-526位點(diǎn)上的基因突變,占16.07%。③1140株陽性標(biāo)本中有140株對(duì)氧氟沙星耐藥,單耐藥率高達(dá)12.28%,是二線藥物中耐藥率最高的一種藥物,而在32株氧氟沙星耐藥的MTB中,所測(cè)的gyr基因突變共有25株,且均為gyrA基因突變,其突變率為78.1%(25/32)。gyrA基因突變株中,以94位點(diǎn)較為常見,占gyrA基因突變的60.0%(15/25),而在gyrB基因上均未發(fā)現(xiàn)突變位點(diǎn)。結(jié)論:①某院就診的結(jié)核患者的結(jié)核分枝桿菌總耐藥率和耐多藥率分別為38.85%和23.68%。②rpoB基因的81bp耐藥區(qū)域上以RRDR-531和RRDR-526兩個(gè)位點(diǎn)上的突變?yōu)橹饕蛔冃问�。③氧氟沙星耐藥與gyr基因突變有一定的關(guān)系,gyrA基因突變是主要突變形式。
[Abstract]:Objective: to investigate the drug resistance of Mycobacterium tuberculosis in patients with tuberculosis, to analyze the mutation of rpoB gyrA and gyrB genes in clinical isolates of Mycobacterium tuberculosis, and to explore the first-line drug rifampicin. The relationship between drug resistance of the second line drug ofloxacin and mutation of rpoB gene gyrA and gyrB gene is a drug resistance study of Mycobacterium tuberculosis (Mycobacterium tuberculosis). The diagnosis and treatment of MDR-TB and XDR-TB) provided a new experimental basis for the diagnosis and treatment of MDR-TB.Methods the specimens of 6535 pulmonary tuberculosis patients in Hunan Provincial chest Hospital were collected. The susceptibility of mycobacterium tuberculosis to Streptomycin (SMC), isoniazid (INHH), rifampicin, EMBB, kanamycin (KMK) and ofloxacin (OFX) OFX was tested by liquid culture and absolute concentration assay (DST). Using Mycobacterium tuberculosis standard strain H37Rv as control, the rifampicin resistant strains were selected from the resistant spectrum of 6 kinds of drugs. The MTB genomic DNA was extracted by ultrasonic lysis method, and the rpoB genes related to rifampicin resistance were amplified. The relationship between gene mutation and drug resistance phenotype was analyzed. 3 ofloxacin resistant strains were selected. The MTB genomic DNA was extracted by ultrasonic lysis method. The gyrA and gyr B genes associated with ofloxacin resistance were amplified and sequenced. Results 1140 cases of Mycobacterium tuberculosis culture positive, 826 cases of initial treatment, 314 cases of re-treatment, 443 cases of drug-resistant pulmonary tuberculosis, including 296 cases of drug-resistant initial treatment, were analyzed. 147 patients were re-treated with drug resistance, the total drug resistance rate was 38.85% / 1140%, the total drug resistance rate of the first treatment patients was 35.84 / 296 / 826, and the total drug resistance rate of the re-treated patients was 46.81% 1477 / 3140.The multidrug resistance rate of 270 patients with multidrug resistance was 23.68,280 / 270,114040%, among which rifampicin was also resistant to rifampicin. Isoniazid and streptomycin accounted for the highest proportion of multidrug resistance patterns, reaching 32.220.87 / 2700.The single drug resistance of rifampicin was the highest, and the rate of single drug resistance was 28.51x / 1140.2 the mutation of rpoB gene in this region was C531T(Ser. 鈫扡eu) was predominant, accounting for 69. 64% of 39 / 56%, of which 29 strains were multidrug resistant strains, followed by gene mutations at the RRDR-526 locus, accounting for 140 of 16. 07% of positive specimens. The rate of single drug resistance was 12. 28%, which was the highest among the second line drugs. Among 32 strains of MTB resistant to ofloxacin, 25 gyr gene mutations were detected, all of which were gyrA gene mutations. The mutation rate was that of 78.1%(25/32).gyrA gene mutants, 94 loci were more common. 60.015 / 25% of the mutation in gyrA gene, but no mutation site was found on the gyrB gene. Conclusion the total drug resistance rate and multidrug resistance rate of Mycobacterium tuberculosis patients in one hospital are 38.85% and RRDR-531 and RRDR-526 in 81bp resistance region of 23.68%.2rpoB gene are 38.85%, respectively. The mutation at two loci was the main mutation form. 3 ofloxacin resistance was related to the mutation of gyr gene, and the mutation of gyra gene was the main mutation form.
【學(xué)位授予單位】：湖南師范大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R440

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本文編號(hào)：1662389