Prevalence and Risk Factors of Primary Drug-Resistant Tuberculosis in China

ObjectiveTo investigatetheprevalence of primary drug-resistant tuberculosis (TB) and associated risk factors in China.We also explored factors contributing tothe transmission of multidrug-resistant tuberculosis (MDR-TB). MethodsA total of 2794 representative,Mycobacterium tuberculosis isolates from treatment-naive patients were subjected to drug susceptibility testing, and risk factors for drug-resistant TBwere analyzed. We also analyzed MDR-TB strain sublineages, drug-resistance-conferring mutations, and risk factors associated with clustered primary MDR strains. ResultsAmong 2794Mycobacterium tuberculosis isolates from treatment-naive patients, the prevalence of any resistance to first-line drugs was 33.2%andthe prevalence of MDR-TB was 5.7%. We did not find any risk factors significantly associated with resistance to first-line drugs.The93 primary MDR-TB isolates were classified into six sublineages, of which, 75 (80.6%) isolates were the RD105-deleted Beijing lineage.The largest sublineage included 65 (69.9%) isolates with concurrent deletions of RD105, RD207, and RD181.Twenty-nine (31.2%) primary MDR strains grouped in clusters;MDR isolates in clusters were more likely to have S531LrpoBmutation. ConclusionThis study indicates that primary drug-resistantTBand MDR-TBstrains are prevalent in China,and multiplemeasures should be taken toaddress drug-resistant TB.     

海南省2014—2016年128株耐多药结核分枝杆菌药敏结果

目的 分析耐多药结核分枝杆菌药物敏感性试验结果，为制定本地区的耐多药结核病治疗方案提供科学依据。方法 收集2014年1月—2016年12月期间海南省结核病医院收治的经痰分离培养药物敏感性试验鉴定为耐多药的菌株共128株，对其结果进行分析。结果 6种抗结核药物耐药率由高到低依次为链霉素（Sm）54.69%，氧氟沙星（Ofx）51.56%，乙胺丁醇（EMB）27.34%，卡那霉素（Km）10.94%，卷曲霉素（CPM）5.47%，丙硫异烟胺（Pto）1.56%；3年间，EMB耐药率差异有统计学意义（P<0.05），2016年EMB耐药率（42.50%）明显高于2014（20.41%）和2015（20.51%）年；3年间Sm、Ofx、Km、CPM、Pto耐药率变化差异均无统计学意义（P>0.05）；耐多药组合模式有18种之多，前3顺位组合模式耐药率依次为（INH+RFP）20.31%，（INH+RFP+Sm）18.75%，（INH+RFP+Ofx）14.84%；前3顺位耐多药种类耐药率依次为耐3种药35.94%，耐4种药25.00%，耐2种药20.31%。结论 海南省耐多药结核病耐药谱广泛且复杂。Km、CPM、Pto耐药率较低，耐多药标准化治疗方案可选价值大。     

A Retrospective Study of Culture-confirmed Mycobacterial Infection among Hospitalized HIV-infected Patients in Beijing,China

A retrospective analysis was performed in two major HIV/AIDS referral hospitals in Beijing to evaluate the prevalence of Mycobacterium tuberculosis (MTB) and non-tuberculous mycobacterial (NTM) infections in HIV-infected patients. A total of 627 patients' data were reviewed, and 102 (16.3%) patients were diagnosed with culture-confirmed mycobacterial infection, including 84 with MTB, 16 with NTM, and 2 with both MTB and NTM. The most frequent clinical complication by mycobacterial infection was pulmonary infection (48/102, 47.1%). The overall rates of multidrug-resistant TB (MDR-TB) and extensively drug-resistant TB (XDR-TB) were 11.9% and 3.4%, respectively. This study underlines the urgent need to intensify screening for mycobacteria coinfection with HIV and to prevent the spread of drug-resistant TB among HIV-infected patients.     

Molecular fingerprinting of multidrug-resistant Mycobacterium tuberculosis strains in Beirut reveals genetic diversity and father to daughter transmission

The typing of six consecutive multidrug-resistant Mycobacterium tuberculosis strains isolated from patients with tuberculosis (TB) at the American University of Beirut Medical Center, was performed by touchdown double-repetitive-element (DRE)-PCR. The isolates exhibited four distinct patterns in DRE-PCR with three isolates exhibiting unique patterns and three isolates yielded similar DNA fragment patterns (cluster pattern). Only two of the three cluster isolates exhibited identical patterns as revealed by restriction fragment length polymorphism (RFLP) targeting specific mutations in the rpoB and katG genes that confer resistance to rifampin and isoniazid, respectively. A direct epidemiological linkage for the two isolates exhibiting genotypic relatedness was also established as the isolates were recovered from a 33-year-old man and his 8-year-old daughter. The data show that transmission of multidrug-resistant M. tuberculosis strains is contributing to the emergence of drug-resistant TB in Beirut. Combining DRE-PCR with RFLP at the rpoB and katG genes could provide a powerful means for investigating the spread of multidrug-resistant M. tuberculosis strains in Lebanon.     

痰耐药结核分枝杆菌rpoB基因的快速检测及利福平耐药机制研究

目的快速检测痰耐药结核分枝杆菌rpoB基因，了解利福平耐药的分予机制。方法根据结核分枝杆菌rpoB基因的耐利福平决定区设计引物．用PCR方法从耐药肺结核患者痰及临床分离菌株扩增rpoB基因片段，对扩增获得的rpoB基因片段进行序列测定，比较分析耐多药、全耐、单耐利福平、全敏感或标准结核分枝杆菌株之间的基因序列差异。结果于6h内从耐药肺结核痰中快速检测到rpoB举因。耐多药菌株、全耐及单耐利福平分离菌株均检测有rpoB基因点突变，突变位点主要为531、516或526常见密码子，且绝大多数为单碱基突变，发现1例MDR-TB出现479位和531位密码子同时突变。敏感株和标准株无基因突变。结论PCR及序列分析方法可快速、准确检测结核分枝杆菌rpoB基岗及其突变，结核分枝杆菌埘利福平的耐药性与rpoB基因点突变有关。     

用绝对浓度法检测结核杆菌药物敏感性试验评价及结果分析

目的 评价结核分支杆菌的药物敏感性，了解目前结核分枝杆菌的耐药状况。方法 采用绝对浓度法对安徽285株结核杆菌进行5种临床常用抗结核药物耐药检测，验证结核杆菌对抗结核药物的敏感性。结果 285株结核分支杆菌的耐药发生率为56．8％，其中单一耐药发生率为11．2％，多重耐药(ivlDR)发生率为45．6％。结论 结核分支杆菌具有很高的MDR发生率，应大力推行结核病全程督导化疗，杜绝不规则用药，减少耐药菌的产生。     

青岛地区结核分支杆菌吡嗪酰胺耐药基因pncA的检测

目的 了解青岛地区结核分支杆菌耐吡嗪酰胺（PZA）分离株pncA基因突变的情况,探讨其与耐PZA之间的关系。方法 应用BACTEC-460培养系统对76例结核分支杆菌临床分离株进行结核分支杆菌常规培养和药敏检测,聚合酶链反应-单链构象多态性（PCR-SSCP）技术检测pncA基因的突变。结果 76株结核分支杆菌临床分离株均经PCR扩增出结核分支杆菌复合群保守序列IS6110基因片段,69株扩增出pncA基因的特异序列。后者包括：SSCP泳动异常者18株,其中耐药株13株,敏感株5株;SSCP泳动正常者51株,其中耐药株14株,敏感株37株。结论 青岛地区结核分支杆菌临床分离株耐PZA主要分子机制之一为pncA基因的突变;PCR—SSCP技术能快速准确地检测结核分支杆菌耐PZA基因pncA的突变,可弥补常规药敏试验的不足。     

Evaluation of Multidrug Resistant Loop-mediated Isothermal Amplification Assay for Detecting the Drug Resistance of Mycobacterium tuberculosis

Objective To evaluate multidrug resistant loop-mediated isothermal amplification (MDR-LAMP) assay for the early diagnosis of multidrug-resistant tuberculosis and to compare the mutation patterns associated with the rpoB, katG, and inhA genes at the Chinese Center for Disease Control and Prevention.Methods MDR-LAMP assay was evaluated using 100 Mycobacterium tuberculosis (Mtb) isolates obtained from the National Reference Laboratory for Tuberculosis in China. Phenotypic resistance to isoniazid and rifampicin and whole-genome sequencing served as reference standards. Results The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of MDR-LAMP were 85.5％, 93.6％, 96.7％, and 74.4％ for the detection of resistance to isoniazid and rifampicin, respectively, and 80.5％, 92.3％, 98.6％, and 41.4％ for the detection of Mtb cultured from smear-positive sputum samples, respectively. When DNA sequencing was used as the reference standard, the sensitivity, specificity, PPV, and NPV of MDR-LAMP were 93.1％, 92.3％, 97.2％, and 82.8％for the detection of katG and inhA gene mutations, respectively, and 89.1％, 88.9％, 93.4％, and 81.1％for the detection of rpoB gene mutation, respectively.Conclusion MDR-LAMP is a rapid and accessible assay for the laboratory identification of rifampicin and isoniazid resistance of Mtb isolates.     

高通量测序筛选结核分枝杆菌耐药株差异基因

目的比较结核分枝杆菌中敏感株、多耐药株、广泛耐药株三者间基因表达的差异,为进一步针对耐药菌株的研究提供方向。方法从深圳市第三人民医院收集敏感株、多耐药株、广泛耐药株各1株,提取菌株RNA,然后构建cDNA文库,使用Genome Analyzer IIx高通量测序仪进行全基因组测序,经过筛选比对后得到测序结果 ,再比较3个菌株间的基因表达差异。结果高通量测序后共得到基因3 968个。将耐药株与敏感株的基因进行比较,选取表达差异高于10倍的基因,筛选得到多耐药株差异基因16个、广泛耐药株差异基因10个。这些表达差异基因大部分与毒素-抗毒素系统、PE/PPE家族蛋白、金属转运蛋白相关。还在多耐药株中发现了一系列高表达的前噬菌体基因及噬菌体整合相关的基因。结论通过高通量测序筛选得到一系列耐药株差异表达基因,为未来对耐药菌株的研究提供依据。     

抗结核新靶点及相关药物的研究进展

肺结核是一种由结核分枝杆菌（Mycobacterium tuberculosis）感染引起的,严重威胁人类健康的传染病。近年来,多药耐药和广泛耐药型结核杆菌的出现给该疾病的控制带来了巨大挑战,在新趋势下,迫切需要作用于新靶点的抗结核药物诞生。本文从结核杆菌细胞生长涉及的必需生物过程、结核杆菌致病和耐药机制3个角度出发,系统综述了抗结核领域新靶点及相关药物的研究进展。     

西安市111例耐多药肺结核患者的药物敏感试验结果

目的 分析西安市目前耐多药结核病（MDR-TB）患者的耐药状况,为合理选用耐多药抗结核方案提供依据。方法 收集西安市2016年1月—2016年12月经痰检分离并鉴定为MDR的结核分枝杆菌药物敏感试验资料,并对其一线及二线抗结核药物进行药敏检测,使用卡方检验对初始耐多药与获得性耐多药组间进行比较。结果 111株MDR的结核分枝杆菌对链霉素和乙胺丁醇的耐药率分别为85.6%和45.0%,二线抗结核药物耐药率由高至低分别为左氧氟沙星（42.3%）、莫西沙星（34.2%）、对氨基水杨酸钠（9.9%）、阿米卡星（8.1%）、丙硫异烟胺（7.2%）、卷曲霉素（4.5%）。链霉素、乙胺丁醇、阿米卡星、卷曲霉素、丙硫异烟胺和对氨基水杨酸钠在初始耐药和获得性耐药两组患者中的耐药率无显著性差异。左氧氟沙星和莫西沙星在获得性耐多药患者中的耐药比例均显著高于初始耐多药组（P<0.05）。结论 西安市MDR-TB临床分离株对链霉素和乙胺丁醇的耐药现象较为严重,对阿米卡星和卷曲霉素敏感性好,而对氟喹诺酮类药物耐药情况严重,在耐多药结核病治疗中应考虑到初始耐药和获得性耐药的不同,制定合理的化疗方案,并强化针对耐多药结核病的控制策略（DOTS-plus策略）,特别是要合理使用氟喹诺酮类药物。     

GeneXpert Mtb/RIF检测技术在结核病诊断中的应用评价

目的 评估GeneXpert Mtb/RIF检测技术对肺结核诊断和耐利福平肺结核筛查的效果,为筛选最佳实验室检测方法提供依据。方法 收集三亚市定点医院结核病门诊2015年5月—2016年12月243例临床诊断为肺结核的痰标本,分别进行涂片镜检、痰培养和GeneXpert Mtb/RIF检测,分析三种方法的敏感度和特异度。以比例法药敏为标准,分析GeneXpert检测利福平耐药的敏感度和特异度。结果 涂片镜检与GeneXpertMtb/RIF对比,GeneXpertMtb/RIF检测结核菌的敏感度为93.7%,特异度为28.9%,GeneXpertMtb/RIF检测与涂片镜检一致率为83.5%。痰培养与GeneXpertMtb/RIF对比,GeneXpert检测结核菌的敏感度为96.7%,特异度为56.6%,GeneXpert检测与痰培养的一致率为91.8%。利福平耐药大多与rpoB基因突变有关,GeneXpertMtb/RIF检测利福平耐药与比例法药敏一致率为98.1%, 敏感度为88.9%,特异度为98.9%,并且可检测出利福平耐药基因位点。结论 GeneXpert Mtb/RIF 检测技术适用于结核分枝杆菌及耐药结核分枝杆菌的快速筛查,在痰涂片和痰培养上补充GeneXpertMtb/RIF检测可提高病原学检出率,尽早发现耐多药病患。     

云南省异烟肼耐药结核分枝杆菌katG和inhA基因突变特征

目的 分析云南省异烟肼耐药结核分枝杆菌katG和inhA基因突变特征。方法 采用PCR方法对云南省异烟肼耐药结核分枝杆菌进行katG和inhA基因扩增,并将扩增产物进行基因测序后比对分析。结果 88株异烟肼耐药菌株中,耐药基因与表型药敏结果的符合率为82.95%(73/88),katG和inhA基因突变率分别为75.00%(66/88)和9.09%(8/88)。最为常见的基因突变位点为katG315(67.05%)和inhA-15(7.95%),katG315位点基因突变主要表现为Ser315Thr(59.09%,52/88)。耐多药与单耐异烟肼菌株中katG基因突变比例,差异有统计学意义(χ²=4.190,P=0.041),而inhA基因及katG315、inhA-15位点基因突变比例,差异无统计学意义(P> 0.05)。结论 云南省异烟肼耐药以katG315和inhA-15基因突变为主,MDR菌株中katG基因突变率高于异烟肼单耐,目前基于基因突变的分子检测技术是检测异烟肼耐药的有效手段。     

结核分枝杆菌耐利福平机制及其研究进展

结核病是严重危害公众健康的全球性公共卫生问题,耐药结核病尤其是耐多药结核病的传播使结核病疫情更加严峻.利福平是重要的一线抗结核药之一,并且作为耐多药结核病的替代标志物.因此了解结核分枝杆菌耐利福平机制对预防及控制耐药结核病有着重要意义.本文就结核分枝杆菌耐利福平机制及其研究进展进行综述.     

抗酸菌耐药结果分析

摘要：目的统计分析评价内蒙古自治区第四医院1997年1月到2021年4月抗酸杆菌阳性菌株耐药情况。方法采用抗酸菌培养加菌型鉴定加药敏试验的方法、基因芯片耐药检测方法、BACTEC MGIT960System960抗酸菌耐药检测方法对抗结核药物异烟肼、利福平、乙胺丁醇、吡嗪酰胺、丙硫异烟胺,左氧氟沙星、氧氟沙星对氨基水杨酸钠、丙硫异胺、卷曲霉素做药物敏感性检测。结果 1612例抗酸菌培养加药敏,其中共检出22例非结核分支杆菌。耐药检测耐药率由高到低分别是异烟肼26.7%>利福平17.3%>链霉素14.6%>乙胺丁醇11.9%>氧氟沙星11.8%>丙硫异烟胺11.5%>耐多药为7.9%>广泛耐药为5.9%>阿米卡星3.1%>卷曲霉素2.4%>对氨基水杨酸钠0.2%; BACTEC MGIT960System 960培养阳性菌株21株,吡嗪酰胺耐药3株,耐药率为1.4%。结论 1 从2017年到2021年,抗酸菌耐药结果显示利福平耐药由历年最低13.3%上升到目前的21%,耐多药由最低0.4%上升到15.3%,氟喹诺酮类氧氟沙星药物抗结核耐药率由0.9%上升到目前26.1%,我院是自治区级治疗结核病的专科医院,一定程度上代表着我区的结核病耐药现状,提示我们结核病控制的复杂性和新的变化。2 耐多药和广泛耐药结核病的存在提示我们要重视结核病的规范治疗,随时评价治疗效果,制定合理有效的抗结核治疗方案。3氟喹诺酮类氧氟沙星药物抗结核耐药率由0.9%上升到目前26.1%,不能除外患者在确诊结核之前应用该类抗生素比率高有关。     

替加环素治疗多或泛耐药鲍曼不动杆菌肺炎的疗效观察

目的探讨替加环素对多或泛耐药鲍曼不动杆菌肺炎的临床疗效。方法回顾性分析2012 年1 月至2013 年8 月因多或泛耐药鲍曼不动杆菌肺炎而使用替加环素治疗的26 例病例资料。观察患者的临床有效率、细菌清除率以及病死率。结果26 例患者使用替加环素, 其中2 例患者使用时间为2 d, 最终24 例纳入临床分析, 其中男18 例, 女6 例; 年龄24 ～81 岁, 平均（ 60. 08 ± 14. 52） 岁。7 例来自外科ICU,17 例来自于内科ICU。所有替加环素均联合给药。替加环素使用时间为3 ～16 d, 平均10 d。5 例患者在治疗期间死亡, 另5 例患者在30 d 内死亡, 30 d 总体病死率为 41. 67% 。12 例患者临床治疗有效, 有效率为50% ; 12 例治疗失败, 失败率为50%; 9 例获得细菌学清除, 细菌清除率为37. 5%。结论替加环素可尝试作为常规抗耐药鲍曼不动杆菌感染治疗方案失败后的抢救性治疗选择, 但尚需要更可靠的临床证据。     

基于常规引物靶向结核分枝杆菌Ⅱ型分枝杆菌散在重复单位的恒温扩增

目的 探讨基于特异常规引物（即引物不需折叠成特定二级结构）靶向恒温扩增重复DNA序列的方法，并测试其检测结核分枝杆菌（Mycobacterium tuberculosis，Mtb）的可能性。方法 以合成的重复DNA或以抽提的细菌基因组DNA为模板，用Bst 2.0 WarmStart DNA聚合酶进行恒温扩增。用琼脂糖凝胶电泳检测扩增结果。结果 合成的重复DNA可用其特异常规引物进行恒温扩增。进一步，Mtb H37Rv Ⅱ型分枝杆菌散在重复单位（mycobacterial interspersed repetitive units，MIRUs）可用其特异常规引物对（即Ⅱ_MIRU-F和Ⅱ_MIRU-R）或单引物（即Ⅱ_MIRU-F）进行恒温扩增。相比于非分枝杆菌菌株、非结核分枝杆菌菌株、Mtb复合物菌株和临床分离株，Ⅱ_MIRU-F能够高特异地扩增Mtb菌株。Ⅱ_MIRU-F针对Mtb H37Rv基因组DNA的检测下限较高，且不能特异地区分Mtb阴性痰液样本和Mtb阳性痰液样本。结论 常规引物可恒温扩增重复DNA序列（包括Mtb Ⅱ型MIRUs）；Mtb Ⅱ型MIRUs特异引物Ⅱ_MIRU-F不适合用于痰液样本的检测。     

结核分枝杆菌痰培养阳性肺结核患者对一线抗结核药耐多药情况分析

目的:分析初治和复治结核分枝杆菌痰培养阳性的肺结核患者对4种一线抗结核药的耐多药(MDR)情况,了解抗结核药耐药现状和形势。方法:选取结核分枝杆菌痰培养阳性患者395例,按中国防痨协会《结核病诊断细菌学检验规程》的要求进行菌种鉴别试验。采用WHO推荐比例法对阳性分离菌株进行药物敏感性检测,计算每种一线药的耐药率及4种一线药的MDR情况。计算初、复治患者的MDR率。结果:4种一线抗结核药物耐药率顺位依次为链霉素(93/395,23.5%),异烟肼(88/395,22.3%),利福平(58/395,14.7%),乙胺丁醇(23/395,5.8%)。初治和复治组患者耐药均以耐链霉素和异烟肼为主,且耐药顺位一致,复治组患者利福平耐药率(17.4%)高于初治组(6.9%)(χ²=6.714,P=0.010)。至少同时对异烟肼和利福平耐药者33例,总MDR率为8.4%,复治患者MDR率(10.2%)高于初治患者MDR率(2.9%)(χ²=5.263,P=0.022)。初、复治组患者MDR形式均以同时耐异烟肼、利福平和链霉素为主,分别为2例(2.0%)和13例(4.4%)。结论:结核分枝菌痰培养阳性肺结核患者耐药频度顺位依次为链霉素、异烟肼、利福平和乙胺丁醇。复治组患者利福平耐药率、MDR率均高于初治组。初、复治组耐多药形式均以同时耐异烟肼、利福平和链霉素为主。     

Hospital outbreak of multidrug-resistant Mycobacterium tuberculosis infections. Factors in transmission to staff and HIV-infected patients.

OBJECTIVE--To describe transmission of multidrug-resistant (MDR) Mycobacterium tuberculosis infection among patients and health care workers (HCWs) in a ward and clinic for human immunodeficiency virus (HIV)-infected patients in a hospital, four studies were conducted. METHODS--Case patients and control patients were persons who had been treated in the HIV ward or clinic, whose clinical course was consistent with tuberculosis and who had at least one positive culture for M tuberculosis between January 1, 1988, and January 31, 1990, resistant to at least isoniazid and rifampin (case patients), or whose isolates were susceptible to all drugs tested (control patients). In the first study, case patients and control patients were compared to identify risk factors for MDR tuberculosis. In the second study, inpatient and outpatient days of MDR tuberculosis case patients were compared to determine whether acid-fast bacillus (AFB) smear-positivity or aerosolized pentamidine use was associated with higher numbers of subsequent MDR tuberculosis cases among exposed patients. In the third study, restriction fragment length polymorphism analysis was performed on available MDR and sensitive M tuberculosis isolates. In the fourth study, skin test conversion rates among HCWs in the HIV ward and clinic were compared with those of HCWs in another ward, and the strength of the associations between skin test conversions among HCWs on the HIV ward and the number of person-days that AFB smear-positive case patients and control patients were on this ward was estimated. RESULTS--Case patients were more likely than control patients to have been exposed on the HIV ward or clinic to an AFB smear-positive case patient (P less than .001). Inpatient and outpatient days of MDR tuberculosis case patients were associated with more subsequent cases of MDR tuberculosis if exposing case patients were smear-positive or if they received aerosolized pentamidine (P less than or equal to .01). Of 13 MDR isolates, all had one of two restriction fragment length polymorphism patterns; 10 sensitive isolates had restriction fragment length polymorphism patterns that were different from each other. The HCW skin test conversion rate was higher on the HIV ward and clinic than on the comparison ward (P less than .01). The risk of occupational acquisition of infection increased in direct proportion to the number of person-days that AFB smear-positive case patients were on the HIV ward (r = .75; P = .005), but did not increase in proportion to the number of person-days that AFB smear-positive control patients were there (r = -.36; P = NS). After isolation measures for AFB smear-positive tuberculosis patients were improved, MDR tuberculosis cases decreased to seven of 214 tuberculosis patients. CONCLUSIONS--Nosocomial transmission of MDR M tuberculosis infection to patients and HCWs occurred on the HIV ward and clinic. Infectiousness of MDR tuberculosis case patients was associated with AFB sputum-smear positivity. Case patients with MDR tuberculosis created a greater risk of skin test conversion for HCWs on the HIV ward than drug-susceptible control patients.