摘要
目的 探究南宁地区结核分枝杆菌(MTB)耐多药表型耐药与基因分子特征的关系,为临床耐药结核病患者的抗结核治疗提供参考.方法 选取2021年3月至2023年7月南宁市第四人民医院收治的300例结核病患者作为研究对象,采集患者的痰液、支气管灌洗液、病理材料或切口分泌物等样本进行菌株培养、体外药物敏感试验,选取同时对利福平和异烟肼表型耐药菌株,或还含有耐氟喹诺酮类药物菌株200株,采用荧光定量聚合酶链式反应(PCR)熔解曲线法进行耐多样基因分子检测,分析菌株的相关基因分子特征.结果 300例患者中,男性占比75.67％,女性占比24.33％;45岁以上的患者占比56.67％;南宁本地户籍占比83.33％;农村人口占比69.00％;复治类型的治疗占比47.67％.在患者的性别(男、女)、年龄(＜30、30～45、＞45岁)、户籍(本地、外地)以及地区(农村、城镇)之间的比较中,差异均有统计学意义(P＜0.05);在初治和复治的比例上,差异无统计学意义(P＞0.05).200例耐多药结核(MDR-TB)菌株表型耐药情况有2种组合,主要是以耐异烟肼+利福平组合为主,比例为53.50％,其次为异烟肼+利福平+喹诺酮类组合,比例为46.50％;与异烟肼表型耐药的相关基因突变率为89.50％,基因突变类型分别为katG315密码子(61.00％),inhA启动子区(-17～-8位点)+katG315密码子(10.00％),ahpC启动子区(-44～-30以及-15～3 位点)(9.00％),inhA 启动子区(-17～-8 位点)(8.00％),ahpC 启动子区(-44～-30 以及-15～3 位点)+katG315密码子(1.00％),ahpC启动子区(-44～-30位点)+katG315密码子(0.50％);与利福平表型耐药的相关基因突变率为 89.50％,分别为 rpoB529～533 位点(50.50％),RrpoB521～528 位点(15.00％),rpoB513～520 位点(12.50％),rpoB513～520+rpoB521～528 位点(11.50％),rpoB507～512+rpoB513～520 位点(2.50％),rpoB521～528+rpoB513～520 位点、rpoB521～528+rpoB529～533 位点(1.50％),ipoB507～512 位点、rpoB507～512+rpoB521～528 位点、rpoB513～520+rpoB529～533位点(0.50％);与喹诺酮类药物表型耐药的相关基因突变率为98.92％,基因突变类型为gyra88～94密码子.结论 南宁地区MTB耐多药形势较为严峻,katG315密码子、inhA启动子区(-17～-8位点)+katG315密码子、rpoB529～533位点、RrpoB521～528位点、gyra88～94密码子是其利福平、异烟肼和耐喹诺酮类药物表型耐药的主要突变位点.
Abstract
Objective To investigate the correlations among multidrug-resistant phenotypic resistance of Mycobacterium tuberculosis(MTB)and gene molecular characteristics in Nanning,and to guide consultation for the anti-tuberculosis treatment of clinically drug-resistant tuberculosis patients.Methods Three hundred tuberculosis patients received in the Fourth Peoples Hospital of Nanning City from during 2021 to July 2023 were picked for the research subjects,and samples of sputum,bronchial lavage fluid,pathological materials or incision secretions were collected for strain culture for in vitro drug sensitivity test,and 200 strains of strains that were resistant to rifampicin and isoniazid at the same time,or also contained fluoroquinolone-resistant strains,were selected.Two hundred strains were selected for in vitro drug sensitivity testing,and 200 strains resistant to both rifampicin and isoniazid,or also containing fluoroquinolone-resistant strains,were analyzed for resistance pheno-types and their related genes and molecular characteristics by using the fluorescence quantitative polymerase chain reaction(PCR)lysis curve method.Results Of the 300 patients,75.67％were male,24.33％were female,56.67％were over 45 years of age,83.33％were of local household registration in Nanning,69.00％were of rural population,and 47.67％were of the relapse type of treatment.In the comparisons be-tween patients'gender(male,female),age(＜30,30-45,＞45 years old),household registration(local,foreign),and region(rural,ur-ban),the differences were statistically significant(P＜0.05);in the proportion of first-time and relapse types of treatment,the differences were not statistically significant(P＞0.05);There were two combinations of drug resistance in 200 cases of multidrug-resistant tuberculosis(MDR-TB)strains,mainly isoniazid+rifampicin combination,with a proportion of 53.50％,followed by isoniazid,rifampicin and quinolones combination,with a proportion of 46.50％;the rate of isoniazid-resistant gene mutations was 89.50％,and the types of gene mutations were katG315 codon(61.00％),inh A promoter region(-17 to-8 sites)+katG315 codon(10.00％),ahpC promoter region(-44 to-30 and-15 to 3 sites)(9.00％),inhA promoter region(-17 to-8 sites)(8.00％),ahpC promoter region(-44 to-30 and-15 to 3 sites)and katG315 codon(1.00％).ahpC promoter region(-44 to-30 loci)and katG315 codon(0.50％);89.50％of mutations in genes associated with rifampicin resistance were at rpoB529 to 533 loci(50.50％),RrpoB521 to 528 loci(15.00％),rpoB513 to 520 loci(12.50％),and rpoB513～520 and rpoB521～528 locus(11.50％),rpoB507～512 and rpoB513～520 locus(2.50％),rpoB521～528 and rpoB513～520 lo-cus,rpoB521～528 and rpoB529～533 locus(1.50％),the rpoB507～512 loci,rpoB507～512 and rpoB521～528 loci,rpoB513～520 and rpoB529～533 loci(0.50％);the mutation rate of the genes related to isoniazid resistance was 98.92％,and the type of the gene mutation was codon gyra88～94.Conclusion The multidrug resistance situation of MTB in Nanning is critical,and katG315 codon,inhA promoter region(-17 to-8 loci)+katG315 codon,rpoB529～533 loci,RrpoB521～528 loci,and gyra88～94 codons are the main rifampicin-resistant,isoniazid-resistant and quinolone-resistant mutation sites.
维普
万方数据