DNA微阵列芯片法检测结核分枝杆菌耐异烟肼基因的研究
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摘要
目的采用DNA微阵列芯片法检测结核分枝杆菌INH耐药基因,为临床诊治提供实验依据。方法应用DNA微阵列芯片法和改良罗氏培养加比例法药敏试验对疑似结核病患者的150份痰液标本进行检测,并对检测的结果进行比较分析。结果 150份痰液标本的涂阳率为60.0%(90/150),DNA微阵列芯片法检测到结核分枝杆菌阳性率为62.7%(94/150),改良罗氏培养法阳性率为60.0%(90/150),比例法药敏试验异烟肼的耐药率为43.9%(36/82),异烟肼的kat G/inh A基因总突变率为35.1%(33/94);以改良罗氏培养加比例法药敏试验为标准,DNA微阵列芯片法检测结核分枝杆菌阳性、异烟肼耐药性和敏感性的符合率分别为97.7%、86.1%和100.0%。结论 DNA微阵列法可快速准确地检测大部分疑似结核病临床标本的kat G和inh A基因突变,可用于临床耐药性的检测从而指导临床用药,也可用于临床诊断中的推广。
Objective To test the resistance isoniazid gene of Mycobacterium tuberculosis using the method of DNAmicroarray chip. Methods A total of 150 sputum specimens of suspected tuberculosis(TB) patients were tested by themethod of DNA microarray chip and the modified Roche medium proportion method,and the results were analyzed andcompared. Results The rate of sputum smear positive was 60.0%(90/150) among the 150 specimens. The positive rate of Mycobacterium tuberculosis was 62.7%(94/150) by the DNA microarray chip method,and the positive rate was 60.0%(90/150)by the modified Roche medium proportion method. The drug resistance rate of isoniazid was 43.9%(36/82) by the drug sensitivetest of the proportion method. The kat G/inh A gene mutation rate of isoniazid was 35.1%(33/94). With the standard of modifiedRoche medium proportion method,the coincidence rates of the Mycobacterium tuberculosis positive rate,isoniazid drugresistance rate,and isoniazid drug sensitive rate tested by the DNA microarray chip were 97.7%,86.1%and 100.0% respectively.Conclusion The DNA microarray method can quickly and reliably detect the gene mutation of kat G and inh A in most clinicalsamples of suspected TB patients. This method can be used to detect clinical drug resistance and guide clinical medication anddiagnosis.
Objective To test the resistance isoniazid gene of Mycobacterium tuberculosis using the method of DNAmicroarray chip. Methods A total of 150 sputum specimens of suspected tuberculosis(TB) patients were tested by themethod of DNA microarray chip and the modified Roche medium proportion method,and the results were analyzed andcompared. Results The rate of sputum smear positive was 60.0%(90/150) among the 150 specimens. The positive rate of Mycobacterium tuberculosis was 62.7%(94/150) by the DNA microarray chip method,and the positive rate was 60.0%(90/150)by the modified Roche medium proportion method. The drug resistance rate of isoniazid was 43.9%(36/82) by the drug sensitivetest of the proportion method. The kat G/inh A gene mutation rate of isoniazid was 35.1%(33/94). With the standard of modifiedRoche medium proportion method,the coincidence rates of the Mycobacterium tuberculosis positive rate,isoniazid drugresistance rate,and isoniazid drug sensitive rate tested by the DNA microarray chip were 97.7%,86.1%and 100.0% respectively.Conclusion The DNA microarray method can quickly and reliably detect the gene mutation of kat G and inh A in most clinicalsamples of suspected TB patients. This method can be used to detect clinical drug resistance and guide clinical medication anddiagnosis.
引文
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[6]林翀,林明冠,廉芳,等.DNA微阵列芯片法检测结核分枝杆菌耐利福平基因的研究[J].中国热带医学,2016,16(2):107-110.
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[9]占玲俊,卢锦标,唐军,等.耐药结核分枝杆菌潜伏-复发感染动物模型的研究进展[J].微生物与感染,2016,11(1):59-64.
[10]张丽霞,邵世峰.结核分枝杆菌异质性所致的基因耐药与表型耐药不一致的研究进展[J].医学综述,2015,14:2526-2528.
[11]陈兆云,蒋静,王家路,等.基因芯片技术在结核耐药基因检测中的应用[J].新疆医学,2012,42(11):55-59.
[12]李桂莲,王撷秀,赵德福.结核分枝杆菌耐药基因的研究进展[J].中国慢性病预防与控制,2006,14(5):377-379.
[13]赵静,王凤平,孙清清,等.DNA微阵列法检测结核分枝杆菌对利福平和异烟肼的耐药性[J].上海交通大学学报医学版,2015,35(11):1651-1654.
[2]沈龙强,梁张,赵桂萍,等.白细胞介素32与结核病的研究进展[J].中国热带医学,2016,16(8):833-835.
[3]QIN N,LI D F,YANG R F.Next-generation sequencing technologies andthe application inmicrobiology[J].Acta Microbiologica Sinica,2011,51(4):445-457.
[4]FLEMING R Y,ZDIGLER S T,WAHON M A,et al.Influence of burn size onthe incidence of contamination of burn wounds by fecal organisms[J].J Burn Care Rehabil,1991,12(6):510.
[5]李史来,陈伟生,黄钥藩,等.1 040株人型结核分枝杆菌耐药性分析[J].中国热带医学,2014,14(6):749-750.
[6]林翀,林明冠,廉芳,等.DNA微阵列芯片法检测结核分枝杆菌耐利福平基因的研究[J].中国热带医学,2016,16(2):107-110.
[7]中国防痨协会基础专业委员会.结核病诊断实验室检验规程[M].北京:中国教育文化出版社,2006.
[8]中华人民共和国国家卫生和计划生育委员会医政医管司.全国临床检验操作规程[M].北京:人民卫生出版社,2015.
[9]占玲俊,卢锦标,唐军,等.耐药结核分枝杆菌潜伏-复发感染动物模型的研究进展[J].微生物与感染,2016,11(1):59-64.
[10]张丽霞,邵世峰.结核分枝杆菌异质性所致的基因耐药与表型耐药不一致的研究进展[J].医学综述,2015,14:2526-2528.
[11]陈兆云,蒋静,王家路,等.基因芯片技术在结核耐药基因检测中的应用[J].新疆医学,2012,42(11):55-59.
[12]李桂莲,王撷秀,赵德福.结核分枝杆菌耐药基因的研究进展[J].中国慢性病预防与控制,2006,14(5):377-379.
[13]赵静,王凤平,孙清清,等.DNA微阵列法检测结核分枝杆菌对利福平和异烟肼的耐药性[J].上海交通大学学报医学版,2015,35(11):1651-1654.