本文選題：廣泛耐藥 + 結(jié)核分枝桿菌��； 參考：《蘇州大學(xué)》2010年碩士論文

【摘要】： 目的篩選XDR-TB臨床分離株,并從耐藥相關(guān)基因突變和藥物外排泵兩方面初步探討XDR-TB耐藥機(jī)制,為開發(fā)XDR-TB的快速診斷技術(shù)和臨床治療方案提供基礎(chǔ)理論支持。 方法利用羅氏藥敏檢測技術(shù)篩選XDR-TB臨床分離株,并檢測XDR-TB分離株的最低抑菌濃度(MIC);提取XDR-TB臨床分離株基因組DNA,PCR擴(kuò)增耐藥相關(guān)基因,測序后分析其突變位點(diǎn);根據(jù)XDR-TB分離株KZN605的15個(gè)特有的SNP位點(diǎn)設(shè)計(jì)引物擴(kuò)增后測序比對;檢測藥物外排泵抑制劑利血平,CCCP,異博定對XDR-TB分離株的MIC值的影響,并通過熒光定量PCR檢測藥物刺激組和未刺激組對編碼藥物外排泵的相關(guān)基因表達(dá)量的影響。 結(jié)果163結(jié)核分枝桿菌(MTB)中篩選得到24株XDR-TB (14.7%),隨機(jī)挑選的10株XDR-TB菌株在rpoB、katG和rpsL均檢測到突變,9株分離株在gyrA檢測到突變,2株在gyrB檢測到突變,6株分離株在rrs檢測到突變,未檢測到tlyA突變,2株檢測到ahpC,3株檢測到inhA,4株檢測到embB;大部分KZN605株特異的SNP位點(diǎn)在本研究均未檢測到;藥物外排泵抑制劑利血平導(dǎo)致1株XDR-TB分離株OFLX的MIC值明顯下降了16倍,其余藥物以及其它分離株的MIC值均無明顯變化;在OFLX刺激后編碼一種ABC運(yùn)輸?shù)鞍椎幕騌v2686大量表達(dá)。 結(jié)論KZN-605的部分SNP位點(diǎn)可能與XDR-TB耐藥無關(guān),耐藥相關(guān)基因突變是XDR-TB分離株耐藥的主要機(jī)制,Rv2686-Rv2687-Rv2688編碼的一種ABC運(yùn)輸?shù)鞍卓赡懿糠謪⑴c了氟喹諾酮類藥物的耐藥產(chǎn)生,gyrB基因D500N和rrs基因A1427G是否與耐藥性相關(guān)還需進(jìn)一步研究探討。
[Abstract]:Objective to screen the clinical isolates of XDR-TB and to explore the mechanism of XDR-TB resistance from two aspects: drug resistance related gene mutation and drug efflux pump. Methods the clinical isolates of XDR-TB were screened by Roche's drug sensitivity detection technique, which provided basic theoretical support for the rapid diagnosis and clinical treatment of XDR-TB. The minimal inhibitory concentration (MIC) of XDR-TB isolate was detected, the gene related to drug resistance was amplified by PCR of genomic DNA of XDR-TB clinical isolate, and its mutation site was analyzed after sequencing, and 15 specific SNP loci of XDR-TB isolate KZN605 were designed for sequencing. The effects of reserpine and verapamil on the MIC of XDR-TB isolates were detected. Fluorescence quantitative PCR was used to detect the effect of drug stimulation group and unstimulated group on the expression of genes related to drug efflux pump. Results 24 strains of XDR-TB were screened out of 163 Mycobacterium tuberculosis (MTB), and 10 strains of XDR-TB were randomly selected. 9 strains were detected in gyrA, 2 strains were detected in GyrB, 6 strains were detected in rrs. No tlyA mutation was detected in 2 strains of tlyA mutation and 4 strains of inhAgna were detected in 4 strains of tlyA, most of the specific SNPs of KZN605 strain were not detected in this study, and reserpine, a drug efflux pump inhibitor, significantly decreased the OFLX value of one XDR-TB isolate by 16 times. After OFLX stimulation, Rv2686, a gene encoding an ABC transport protein, was highly expressed. Conclusion some SNP loci of KZN-605 may be independent of XDR-TB resistance. The mutation of drug-resistance-related genes is the main mechanism of drug resistance in XDR-TB isolates. An ABC transport protein encoded by Rv2686-Rv2687-Rv2688 may be partly involved in the production of fluoroquinolones resistance and whether D500N and A1427G of rrs gene are related to drug resistance should be further studied.
【學(xué)位授予單位】：蘇州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2010
【分類號(hào)】：R378.911

【參考文獻(xiàn)】

相關(guān)期刊論文 前5條

1 付佑輝;鈄方芳;鄭瑞娟;王潔;金文國;胡忠義;;MODS技術(shù)快速檢測結(jié)核分枝桿菌左氧氟沙星耐藥性研究[J];臨床肺科雜志;2009年02期

2 崔振玲,景奉香,胡忠義,吳自榮,趙建龍;基因芯片檢測結(jié)核分枝桿菌利福平和異煙肼耐藥性研究[J];中華結(jié)核和呼吸雜志;2004年07期

3 王u&民;劉宇紅;姜廣路;劉洋;付育紅;端木宏謹(jǐn);;世界衛(wèi)生組織中國結(jié)核病耐藥監(jiān)測的結(jié)果評價(jià)[J];中華檢驗(yàn)醫(yī)學(xué)雜志;2007年08期

4 李衛(wèi)民,端木宏謹(jǐn),王黎霞,丁北川,劉宇紅,李傳友,傅喻;2000年中國結(jié)核病流行病學(xué)抽樣調(diào)查菌株分子流行病學(xué)特征[J];中華醫(yī)學(xué)雜志;2003年14期

5 崔振玲;王潔;陸俊梅;汪小黎;胡忠義;;結(jié)核分枝桿菌臨床分離株對三種氟喹諾酮類藥物的敏感性分析研究[J];中華臨床醫(yī)師雜志(電子版);2009年12期

，

本文編號(hào)：1998955