本文選題：結(jié)核分枝桿菌 + RD105基因缺失��； 參考：《吉林大學(xué)》2015年碩士論文

【摘要】：目的了解22株結(jié)核分枝桿菌臨床分離株的基因型分布情況，探索適合結(jié)核分枝桿菌北京基因型鑒定的方法。探討結(jié)核分枝桿菌北京基因型菌株與耐藥是否相關(guān)。分析結(jié)核分枝桿菌耐藥性與耐藥基因之間的聯(lián)系。方法對(duì)收集的吉林省22株結(jié)核分枝桿菌臨床分離株進(jìn)行RD105缺失基因檢測(cè)法基因分型研究。應(yīng)用絕對(duì)濃度法藥物敏感試驗(yàn)方法檢測(cè)22株結(jié)核分枝桿菌對(duì)異煙肼、利福平、鏈霉素、卡那霉素、氧氟沙星及對(duì)氨基水楊酸六種抗結(jié)核藥物的耐藥性，最后對(duì)藥敏結(jié)果和基因分型結(jié)果進(jìn)行分析。對(duì)于耐藥菌株將其katG基因、rpoB基因、rpsl基因、rrs基因、gyrA基因及thyA基因進(jìn)行測(cè)序，檢測(cè)其耐藥基因是否存在突變并且確定突變的類型。結(jié)果22株結(jié)核分枝桿菌臨床分離株中有10株耐藥菌株，占45.5%。對(duì)利福平耐藥的有6株，占27.3%；對(duì)異煙肼耐藥的有4株，占18.0%；對(duì)鏈霉素耐藥的有2株，占9%；對(duì)卡那霉素耐藥的有4株，占18.0%；對(duì)氧氟沙星耐藥的有3株，占13.6%，對(duì)對(duì)氨基水楊酸耐藥的有一株，占4.5%，北京基因型菌株占77.3%(17/22)。17株北京家族結(jié)核分枝桿菌臨床分離株中52.9%（9/17）表現(xiàn)對(duì)六種抗結(jié)核藥物敏感，47.1%（8/17）表現(xiàn)為耐藥；非北京家族菌株中60%（3/5）表現(xiàn)為敏感，40%（2/5）表現(xiàn)為耐藥。經(jīng)檢驗(yàn)，北京家族菌株與非北京家族菌株耐藥率差異無(wú)統(tǒng)計(jì)學(xué)意義。4株異煙肼耐藥株中，有1株未發(fā)生katG基因突變，其它3株katG基因發(fā)生突變，均發(fā)生在315位點(diǎn)，由AGC（Ser）突變成ACC（Thr）。6株利福平耐藥株中，有4株rpoB基因發(fā)生531位點(diǎn)突變，由TCG（Ser）突變成TTG（Leu）。2株rpoB基因發(fā)生526位點(diǎn)突變，由CAC（His）突變成GAC（Asp）。2株鏈霉素耐藥株rpsl基因均發(fā)生43位點(diǎn)突變，由AAG（Lys）突變成AGG（Arg）。4株卡那霉素耐藥株中，有1株rrs基因未發(fā)生突變，其它3株rrs基因均發(fā)生11位點(diǎn)突變，由GGT（Gly）突變成GTT（Val）。3株氧氟沙星耐藥株gyrA基因均發(fā)生94位點(diǎn)突變，GAC（Asp）突變成GCC（Gly）。1株對(duì)氨基水楊酸耐藥株thyA基因在355位點(diǎn)C堿基缺失，由CTG（Leu）突變成GTC（Val）。結(jié)論RD105缺失基因檢測(cè)法是鑒定MTB北京基因型的簡(jiǎn)便、有效的方法；北京基因型菌株為所研究的結(jié)核分枝桿菌株中的優(yōu)勢(shì)菌株；北京基因型菌株與非北京基因型菌株兩者耐藥率的差別無(wú)統(tǒng)計(jì)學(xué)意義。結(jié)核分枝桿菌對(duì)抗結(jié)核藥物耐藥與相關(guān)耐藥基因突變密切相關(guān)。
[Abstract]:Objective to investigate the genotypic distribution of 22 clinical isolates of Mycobacterium tuberculosis and to explore a suitable method for identification of mycobacterium tuberculosis in Beijing. To investigate whether the Beijing genotype of Mycobacterium tuberculosis is related to drug resistance. To analyze the relationship between drug resistance and drug resistance genes of Mycobacterium tuberculosis. Methods 22 clinical isolates of Mycobacterium tuberculosis collected from Jilin Province were genotyped by RD105 deletion method. The drug resistance of 22 strains of Mycobacterium tuberculosis to isoniazid, rifampicin, streptomycin, kanamycin, ofloxacin and p-aminosalicylic acid was detected by absolute concentration method. Finally, the results of drug sensitivity and genotyping were analyzed. The katG gene, rpsl gene, rrs gene, gyrA gene and thyA gene were sequenced to detect whether the drug resistance gene had mutation and determine the type of mutation. Results among the 22 clinical isolates of Mycobacterium tuberculosis, 10 strains were drug-resistant, accounting for 45.5%. Six strains were resistant to rifampicin, accounting for 27.3um; four strains were resistant to isoniazid, accounting for 18.0%; two strains were resistant to streptomycin (9 strains); four strains were resistant to kanamycin (18.0%); three strains were resistant to ofloxacin, One strain was resistant to aminosalicylic acid, accounting for 4.5%, and the Beijing genotype strain accounted for 77.3% 22 / 22 .17 strains of Mycobacterium tuberculosis in the family of Beijing. 52.9% of the 17 strains showed resistance to the six anti-tuberculosis drugs (47.1% / 87%). 60 / 5 of the non-Beijing strains showed resistance to drugs. The results showed that there was no significant difference in drug resistance rate between Beijing family strains and non-Beijing family strains. Of the 4 isoniazid resistant strains, 1 strain had no katG gene mutation, while the other 3 strains had katG gene mutation, all of which occurred at 315 locus. Among the rifampicin resistant strains, there were 531 mutation in rpoB gene, 526 site mutation in rpoB gene in TTG(Leu).2 strain, 43 locus mutation in rpsl gene of streptomycin resistant strain GAC(Asp).2. The rrs gene was not mutated in one of the kanamycin resistant strains of AGG(Arg).4, and 11 loci were found in all the other three strains of rrs gene. The 94 locus mutation of the gyrA gene of GTT(Val).3 strain was mutated to the deletion of thyA gene at 355 locus from GCC(Gly).1 strain. Conclusion the detection of RD105 deletion gene is a simple and effective method to identify the Beijing genotype of MTB. There was no significant difference in drug resistance between Beijing genotype strains and non-Beijing genotype strains. Anti-TB drug resistance of Mycobacterium tuberculosis is closely related to the mutation of related drug resistance genes.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R446.5

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