河南省耐多药结核分枝杆菌对吡嗪酰胺耐药性分析及其与耐药相关基因pncA突变的关系  

Analysis of pyrazinamide resistance of multidrug-resistant Mycobacterium tuberculosis in Henan Province and its relationship with the mutation of drug resistance-related gene pncA

作　　者：张国龙 Zhang Guolong(Tuberculosis Control Institute,Henan Provincial Center for Disease Control and Prevention,Zhengzhou 450016,China)

机构地区：[1]河南省疾病预防控制中心结核病防治所,郑州450016 [2]河南省疾病预防控制中心性病艾滋病防治所,郑州450016

出　　处：《中华传染病杂志》2021年第7期424-429,共6页Chinese Journal of Infectious Diseases

摘　　要：目的评价耐药相关基因pncA突变检测河南省耐多药结核病患者吡嗪酰胺耐药性的效果。方法收集2018年1月至12月于河南省10个结核病耐药监测地区就诊的152例耐多药结核病患者的痰标本。对患者进行问卷调查,收集年龄、性别、治疗史、痰培养等信息。最终将问卷及菌株标本送至河南省疾病预防控制中心进行后续研究。采用结核分枝杆菌液体培养进行吡嗪酰胺药物敏感试验。采用聚合酶链反应和基因测序鉴定pncA的突变情况,并与标准菌株H37Rv的序列进行比较。分析吡嗪酰胺耐药表型与抗结核治疗效果的相关性。统计学分析采用χ^(2)检验和独立样本t检验。结果152株耐多药结核分枝杆菌菌株中,105株为吡嗪酰胺表型耐药菌株。对异烟肼、利福平、乙胺丁醇和链霉素全部耐药的菌株,以及准广泛耐药菌株和广泛耐药菌株对吡嗪酰胺的耐药率均较高,分别为80.39%(82/102)、81.13%(43/53)和92.59%(25/27)。102株菌株的pncA基因发生突变。以表型药物敏感试验结果为参照,pncA基因突变检测吡嗪酰胺耐药性的灵敏度为89.52%(95%可信区间81.64%~94.39%),特异度为89.36%(95%可信区间76.10%~96.01%)。共存在100种不同类型的pncA突变,包括80种点突变和20种插入或缺失突变,其中有13株菌株存在多个位点突变。7株菌株pncA启动子发生突变,分别为-7G插入突变、-11T到C点突变和-12A到G点突变,3组菌株中pncA mRNA相对表达量分别为0.21±0.05、0.31±0.08和0.33±0.03,均低于标准株H37Rv(设置其pncA mRNA表达量为1.00),差异均有统计学意义(t=4.57、2.43、3.65,均P<0.05)。治疗后不同时间段吡嗪酰胺耐药患者和敏感患者的痰菌阴转率差异有统计学意义(χ^(2)=10.01,P=0.02),其中吡嗪酰胺耐药患者治疗6个月末的阴转率为1.08%(1/93),而吡嗪酰胺敏感患者为7.14%(3/42)。结论耐多药结核分枝杆菌呲嗪酰胺耐药与pncA基因突变相关,该突变散在�Objective To evaluate the performance of the sequencing method of pncA mutations in detecting pyrazinamide(PZA)resistance in multidrug-resistant tuberculosis(MDR-TB)patients in Henan Province.Methods Sputum samples of 152 MDR-TB patients were collected from 10 drug-resistance surveillance areas in Henan Province from January to December 2018.Questionnaire survey was conducted to collect the information,such as age,gender,treatment history and sputum culture results.The questionnaire and strain samples were sent to Henan Provincial Center for Disease Control and Prevention for further study.PZA susceptibility test was performed using the liquid medium for Mycobacterium tuberculosis.The mutations of pncA were detected by polymerase chain reaction(PCR)and sequencing,then compared with the sequence of standard strain H37Rv.The association between PZA resistant phenotype and treatment outcomes was also investigated.Chi square test and independent sample t test were used for statistical analysis.Results Among 152 MDR-TB isolates,105 showed phenotypically PZA resistant.The proportion of PZA resistance in the isolates with isoniazid,rifampicin,ethambutol and streptomycin resistance,pre-extensively drug-resistant and extensively drug-resistant(XDR)were 80.39%(82/102),81.13%(43/53)and 92.59%(25/27),respectively.One hundred and two isolates had mutations in the pncA gene.Based on the results of the phenotypic drug sensitivity test,the sensitivity of pncA gene mutation detection for PZA resistance was 89.52%(95%confidence interval(CI)81.64%-94.39%),and the specificity was 89.36%(95%CI 76.10%-96.01%).These MDR-TB isolates harbored 100 different mutation patterns in the pncA gene,including 80 point mutations and 20 indel mutations,and 13 isolates harbored multiple mutations.Seven strains had mutation in the promoter of pncA,including-7G insertion,-11T to C and-12A to G.The relative expression levels of pncA mRNA in the three groups were 0.21±0.05,0.31±0.08 and 0.33±0.03,respectively,which were all lower than that of H37Rv(1

关 键 词：分枝杆菌 结核 吡嗪酰胺 结核 抗多种药物性 pncA基因 

分 类 号：R52[医药卫生—内科学]