958例结核病患者临床分离株耐药基因检测结果分析

doi:10.3969/j.issn.1000-6621.2019.05.013

摘要/Abstract

摘要：

目的分析958例患者结核分枝杆菌的相关基因突变位点。方法收集2017年1—12月西安市胸科医院对痰液、组织、胸腹腔积液等各类型临床标本进行分枝杆菌培养阳性的2718例患者的菌液,经菌种鉴定非结核分枝杆菌感染患者41例,结核分枝杆菌感染患者2677例,选择结核分枝杆菌菌种同时进行MGIT 960液体药物敏感性试验(简称“药敏试验”)和耐药基因检测的患者958例,分析不同性别、年龄组患者的结核分枝杆菌菌株间突变位点差异及突变位点与表型药敏试验检测结果的符合率。计数资料采用χ²检验,以P<0.05为差异有统计学意义。结果 958例患者菌株的基因突变位点分析中,任一位点突变率为19.52%(187/958),5~、21~、41~、61~83岁各年龄组的菌株ropB基因位点突变率分别为7.89%(9/114)、13.52%(63/466)、18.27%(38/208)和9.41%(16/170)。不同年龄组中,中年组的ropB基因位点突变率与青少年组和老年组比较差异有统计学意义(χ²值分别为6.358和5.994,P值分别为0.013和0.018);男性任一位点突变率为19.45%(121/622),女性为19.05%(64/336),不同性别比较差异无统计学意义(χ²=0.023,P=0.932);突变位点与表型药敏试验的符合率以利福平ropB 531(C-T)(96.10%,74/77)、异烟肼katG 315(G-C)(98.45%,127/129)最高,但ropB 511(T-C)与利福平表型药敏试验结果相符率较差(0.00%,0/10)。结论不同年龄组患者的菌株ropB基因位点突变率差异有统计学意义,中年组突变率最高;不同性别患者的突变率差异无统计学意义;应当结合表型药敏试验的检测结果进行耐药基因位点突变在诊断意义上的分析。

关键词: 分枝杆菌, 结核, 微生物敏感性试验, 分子探针技术, 点突变, 对比研究

Abstract:

Objective Analysis of mutation sites of drug resistance genes in 958 M.tuberculosis clinical isolates.Methods The positive mycobacterial cultures of 2718 patients were collected from various clinical specimens such as sputum, tissue, and pleural effusion in Xi’an Chest Hospital from January to December 2017. Forty-one cases of non-tuberculous mycobacterial infection and 2677 cases of Mycobacterium tuberculosis infection were distinguished by strains identification.The information including results of M.tuberculosis cultures and drug susceptibility test (“drug sensitivity test”) by MGIT 960 liquid and drug resistance test by gene chip in 958 patients were collected.The coincidence rate of mutation sites and phenotypic drug susceptibility test results were analyzed. The count data were analyzed by χ² test, and P<0.05 was considered statistically significant.Results The mutation rate of a single site was 19.52% (187/958) in all of the clinical isolates.The ropB gene mutation rates in the 5-, 21-, 41-, 61-83 age groups were 7.89% (9/114), 13.52% (63/466), 18.27% (38/208), and 9.41% (16/170), respectively. The mutation rate of the ropB in the middle-aged group was statistically different from that in the adolescent group (χ²=6.358, P=0.013) and the elderly group (χ²=5.994, P=0.018). The mutation rate at any one site was 19.45% (121/622) in the male population and 19.05% (64/336) in the female population.There was no significant difference between the male and female groups (χ²=0.023, P=0.932). There was a higher coincidence rate in mutation of rifampicin ropB 531 (C-T) (96.10%, 74/77) and isoniazid katG 315 (G-C) (98.45%, 127/129) and the phenotypic susceptibility. However, ropB 511 (T-C) was inferior coincidence rate to rifampicin phenotype (0.00%,0/10).Conclusion T here was a statistically significant difference in the mutation rate of ropB gene locus in patients of different age groups.The mutation rate of ropB gene happened in the middle-aged group was the highest. There was no significant difference in the mutation rate between the different gender groups.Diagnosis should be done by drug gene mutation combined with phenotypic susceptibility.

Key words: Mycobacterium tuberculosis, Microbial sensitivity tests, Molecular probe techniques, Point mutation, Comparative study

崔晓利,杨翰,邬霞,郑丹,尤振宇,党丽云. 958例结核病患者临床分离株耐药基因检测结果分析[J]. 中国防痨杂志, 2019, 41(5): 541-545. doi: 10.3969/j.issn.1000-6621.2019.05.013

Xiao-li CUI,Han YANG,Xia WU,Dan ZHENG,Zhen-yu YOU,Li-yun. DANG. Analysis of drug susceptibility and drug resistance gene locus mutation in 958 tuberculosis patients[J]. Chinese Journal of Antituberculosis, 2019, 41(5): 541-545. doi: 10.3969/j.issn.1000-6621.2019.05.013

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参考文献 13

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耐药基因	突变位点	突变例数	突变率 (%)
任一突变率		187	19.52
利福平耐药基因
ropB	531(C-T)	77	8.04
	526(C-G)	15	1.57
	511(T-C)	10	1.04
	516(A-T)	6	0.63
	526(A-T)	5	0.52
	533(T-C)	3	0.31
	513(C-A)	2	0.21
	516(G-T)	2	0.21
	531(C-G)	2	0.21
	511(T-C)&516(A-G)	1	0.10
	511(T-C)&526(C-T)	1	0.10
	513(C-A)&526(C-G)	1	0.10
	526(A-G)	1	0.10
异烟肼耐药基因
katG	315(G-C)	129	13.47
	315(G-A)	6	0.63
inhA	15(C-T)	32	3.34

突变类型	性别		χ²值	P值	年龄组(岁)				χ²值	P值
突变类型	男	女	χ²值	P值	5~	21~	41~	61~83	χ²值	P值
任一突变	19.45 (121/622)	19.05 (64/336)	0.023	0.932	12.28 (14/114)	17.38 (81/466)	26.44 (55/208)	20.59 (35/170)	11.695	0.009
ropB突变	14.15 (88/622)	11.31 (38/336)	1.539	0.230	7.89 (9/114)	13.52 (63/466)	18.27 (38/208)	9.41 (16/170)	0.246	0.041
katG突变	13.67 (85/622)	14.58 (49/336)	0.153	0.697	8.77 (10/114)	13.30 (62/466)	17.31 (36/208)	15.29 (26/170)	5.234	0.155
inhA突变	3.86 (24/622)	2.38 (8/336)	1.475	0.262	4.39 (5/114)	2.58 (12/466)	3.85 (8/208)	4.12 (7/170)	-	0.400

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