脊柱结核个体化药物治疗的临床效果观察

目的 探讨药敏试验指导下的脊柱结核个体化药物治疗的临床效果.方法 选择2005年8月至2010年1月诊治的132例脊柱结核手术患者进行结核分枝杆菌培养和药敏试验,随访超过12个月者62例纳入本研究,其中男性37例,女性25例;年龄4～67岁,平均33.6岁.患者均接受手术治疗,术中收集脓液、干酪样组织,常规处理后接种培养液,使用BACT/ALERT 3D细菌自动培养分析系统快速培养,培养阳性者采用改良罗氏培养液按绝对浓度法进行药敏试验,并根据结果制定抗结核化疗方案.术后1、3、6、9、12个月复查各1次,以后每6个月复查1次,观察临床表现及红细胞沉降率、X线片、三维CT、MRI的变化,分析局部及全身结核病转归、术区骨性融合等情况.结果 检测耗时28～58 d,平均42 d;培养阳性率45.2%(28/62).耐药率达24.2%,其中异烟肼12.9%、利福平4.8%、乙胺丁醇3.2%、链霉素9.7%、帕司烟肼6.4%、左氧氟沙星14.5%、利福喷汀1.6%.随访时间12～44个月,平均21个月.术后2周内切口愈合率98.4%(61/62).椎间植骨均获Ⅰ级骨性融合,融合时间8～12个月.结论 耐药结核形势严峻,药敏试验指导下的脊柱结核个体化药物治疗是减少复发、复治的关键.     

脊柱结核不同技术与病理标本结核分枝杆菌培养及药敏试验的比较

[目的]探讨不同技术与脊柱结核病理标本结核分枝杆菌培养及药敏试验的阳性结果。[方法]采用后前路、前路、后路手术方式获取262例脊柱结核患者的病理标本:脓液、肉芽组织、病灶壁,分别采用Bac T/ALERT3D法和改良罗氏培养法进行结核分枝杆菌培养,并对培养阳性细菌进行药敏试验。比较两种培养方法及三种病理组织标本各自的阳性率,分析结核杆菌耐药情况。[结果]Bac T/ALERT 3D法和改良罗氏培养法培养结核分枝杆菌的阳性率分别为32.82%和20.99%。两者比较差异有统计学意义(P<0.05);Bac T/ALERT3D法中脓液、肉芽组织、病灶壁三种标本的阳性率分别为34.17%、23.08%、15.58%,差异有统计学意义(P<0.05);改良罗氏培养法中上述三种标本的阳性率分别为26.13%、12.43%、5.84%,差异有统计学意义(P<0.05);对78份结核菌培养阳性的结核菌落分别进行抗痨药物异烟肼、利福平、链霉素、吡嗪酰胺、乙胺丁醇的药物敏感性测定,药物敏感率分别为87.18%、89.74%、98.72%、98.72%、97.44%,耐药率分别为12.82%、10.26%、1.28%、1.28%、2.56%。[结论]与改良罗氏培养法相比,Bac T/ALERT3D液体培养系统具有明显缩短检出时间、提高检出率的优势,对结核病的早期诊断和治疗阶段的疗效评估等方面更具有应用价值;脓液标本培养过程简单,且其培养阳性率高于结核肉芽组织及病灶壁,可作为脊柱结核标本培养的首选方法。     

浙江省耐利福平结核分枝杆菌rpoB基因突变研究

目的 了解浙江省耐利福平结核分枝杆菌rpoB基因突变特征.方法 利用传统药敏检测法和以PCR为基础的DNA测序对从浙江省立同德医院和浙江省中西医结合医院的188例结核病患者中分离的188株结核分枝杆菌进行耐药分析和rpoB基因突变分析,观察结核分枝杆菌利福平耐药决定区基因突变情况与临床耐药的关系.结果 188株临床分离株中有57株为耐药株(30.3％),其中单耐利福平18株(9.6％),单耐其他抗结核药物28株(14.9％)(异烟肼10株、链霉素12株和乙胺丁醇6株);耐2种或2种以上药物(包括异烟肼、链霉素和乙胺丁醇)的多耐药临床分离株11株(5.9％).29株耐利福平菌株中,27株(93.1％)存在rpoB基因突变,其中526位突变率为55.6％ (16/27),513位突变率为22.2％(5/27),531位突变率为14.8％ (4/27),529位突变率为7.4％ (2/27);28株耐其他抗结核药物分离株中,4株(14.3％)存在rpoB基因突变,突变位点分别位于526位(2株)和513位(2株).其余131株敏感菌株rpoB基因均无突变发生.结论 浙江省耐利福平结核分枝杆菌与rpoB基因突变有关,主要以526位与513位突变为主.     

骨关节结核病灶中耐多药结核分枝杆菌对疗效的影响

目的回顾性探索骨关节结核病灶中耐多药结核分枝杆菌对骨关节结核病疗效的影响及其对策。方法1993至1999年,确诊为复治骨关节结核病250例,男143例,女107例;年龄2～72岁,平均25.44岁;病变在脊椎、髋、膝和其他小关节。穿刺或外科手术获得病灶中脓液、干酪样物或肉芽组织等为标本,以BACTECTB460检测仪进行结核菌培养和药敏试验,NAP试剂进行初级鉴定,NTM分离培养物进一步鉴定分枝杆菌菌种。结果250例标本中,分离培养阳性培养物48例。结核分枝杆菌46例,阳性率为18.4%。其中同时耐异烟肼(INH)、利福平(RFP)、链霉素(SM)和乙胺丁醇(EMB)中三种药的结核分枝杆菌4例,同时耐异烟肼、利福平、链霉素和乙胺丁醇11例,共15例(6%)。在15例中有8例此次手术后切口破溃形成窦道,依据药敏试验结果改变化疗方案治疗,并经前后2或3次再手术,7例治愈,1例因窦道继发感染而死亡。另2例为非结核分枝杆菌(NTM),1例鉴定为鸟胞内分枝杆菌复合群,另1例鉴定未有结果,但均为耐多药(INH、RFP、SM、EMB、KM、CS)的NTM,治疗效果差。结论骨关节结核病灶中发现耐多药结核分枝杆菌(6%)和非结核分枝杆菌的病例,疗效极差,此类病例的化疗方案应个体化,可能需要多次手术才能治愈。     

288份分枝杆菌菌型鉴定及药敏试验结果监测分析

目的:通过分析石家庄市第五医院肺结核病人的分枝杆菌菌种鉴定及药敏试验结果,了解耐药情况,为临床治疗肺结核病人提供可靠的实验数据.方法:对2007～2010年就诊于我院的肺结核病人的1001份标本,用改良罗氏法培养,对288份分离培养阳性菌株进行菌型鉴定及药敏试验,试验结果进行统计分析.结果:①培养阳性率295/1001(29.5％),288例培养阳性标本,人型结核分枝杆菌263例,牛型分枝杆菌28例,乌型分支杆菌1例,其他非典型分枝杆菌3例.②耐药菌株98株,人型分支杆菌耐药比例低于牛型(30.4％Vs57.1％,P＜0.01).耐药率从高到底依次为:乙胺丁醇18.4％、链霉素15.3％、利福平14.2％、异烟肼12.2％、氧氟沙星9.0％、左氧氟沙星8.3％、力克肺疾6.9％、阿米卡星5.8％、利福布丁4.5％.③单耐药菌株20例(7.6％)多耐药菌株25例(8.7％),耐多药菌株22例(7.6％),广泛耐药结核菌株6例(2.1％).结论:结核分支杆菌对抗结核药耐药率高,临床应根据结核杆菌药敏试验的结果,合理使用抗结核药物,减少耐药菌株的发生.     

脊柱结核耐药性检测及耐药基因PCR-SSCP分析的应用价值

目的了解临床脊柱结核耐药情况,探讨耐药基因PCR-SSCP分析的临床应用价值。方法31例脊柱结核病灶应用BACTECMGIT960培养,所得临床分离株行药敏试验,对耐药基因rpsL、katG、rpoB行PCR-SS-CP分析。结果31例样本中27例培养阳性,其中18株存在不同程度耐药,总耐药率为66.67%。药物耐药率由高至低为链霉素10株(55.56%)、异烟肼8株(44.44%)、利福平7株(38.89%)、PZA3株(16.67%)。耐链霉素株rpsL突变率为70%,耐异烟肼株katG突变率为50%,耐利福平株rpoB突变率为71.43%。高浓度水平耐药株耐药基因突变率明显高于低浓度水平耐药株。结论常规药敏试验与耐药基因分析相结合可能更能准确地反应MTB的耐药情况。     

获得性免疫缺陷综合征合并结核病患者对结核分枝杆菌二线药物耐药特征分析

目的了解获得性免疫缺陷综合征（AIDS）合并结核病（TB）患者感染结核分枝杆菌二线药物耐药特点。 方法选取2010年4月至2012年10月于北京大学地坛医院教学医院住院的艾滋病合并结核病患者标本,由中国疾病预防控制中心培养鉴定。进行4种一线药物（异烟肼、利福平、链霉素、乙胺丁醇）和4种二线药物（卷曲霉素、卡那霉素、氧氟沙星、乙硫异烟胺）药敏试验监测,并对所有菌株在gyrA、gyrB、rrs、tlya、eis和ethA基因位点进行DNA测序以检测基因多态性。 结果经培养鉴定共得到31株结核分枝杆菌,其中12株耐卷曲霉素,8株耐氧氟沙星,4株耐卡那霉素,5株耐乙硫异烟胺,耐药率分别为38.71%、25.81%、12.90%和16.13%。7株菌为耐多药菌株,1株菌为广泛耐药菌株,耐药率分别为22.58%和3.23%。耐药菌株最常见的突变位点是rrs₁₄₀₁,gyrA₉₄和gyrA₉₀。一线敏感菌株中氧氟沙星的耐药率显著低于一线耐药菌株（P = 0.012）。性别与结核分枝杆菌耐药差异无统计学意义（P = 0.533）,年龄> 40岁组的氧氟沙星耐药率低于其余两组（P = 0.043）。结核初治组与复治组患者二线耐药率、CD4水平差异无统计学意义（P = 0.333、0.307）。 结论AIDS合并TB患者存在二线抗结核药物原发耐药,其中卷曲霉素耐药率最高,其次是氧氟沙星。     

脊柱结核脓液中结核分枝杆菌耐利福平基因检测的实验研究

目的:探讨脊柱结核脓液中结核分枝杆菌对利福平(RFP)的耐药机制;评价聚合酶链反应-单链构象多态性(polymerasechainreactionsingle-strandconformationpolymorphism,PCR-SSCP)在脊柱结核检测耐RFP基因中的临床应用价值。方法:将脊柱结核病灶中的脓液经预处理后接种于改良罗氏培养基上,对所培养出的菌株应用PCR-SSCP进行检测。检测包括核心区域(耐药决定区域)在内的rpoB基因。结果:PCR-SSCP检测耐RFP菌株的灵敏度和特异性均为100%,与常规经典药物敏感试验无差异(P<0.05),但检测时间明显缩短,仅用2d。结论:rpoB基因突变是结核分枝杆菌对RFP耐药的重要机制,应用PCR-SSCP检测具有较高的灵敏性、特异性及快速性,是一种对脊柱结核耐RFP菌株快速检测的方法。     

应用Xpert MTB/RIF对脊柱结核临床标本行结核分枝杆菌与利福平耐药性检测的验证性研究

 目的 采用Xpert MTB/RIF系统对系列脊柱结核临床标本进行结核分枝杆菌检出与利福平耐药基因rpoB突变检测,初步验证该项技术的可行性与准确性。方法 自全军结核病研究所标本库中筛选140份脊柱结核临床标本,对标本行前处理后采用Xpert MTB/RIF系统进行结核分枝杆菌与利福平耐药基因rpoB的突变检测,以培养结果及表型药敏试验为金标准,判断Xpert MTB/RIF检测的敏感度、特异度、95%置信区间及检测耗时。结果 对临床确诊为脊柱结核的140份临床标本,Xpert MTB/RIF系统的结核分枝杆菌阳性检出率为63.57% (89/140);在64份培养阳性标本中,Xpert MTB/RIF检测结核分枝杆菌的敏感度为98.44% (63/64);在76份培养阴性标本中,Xpert MTB/RIF检测结核分枝杆菌的敏感度为34.21% (26/76)。以表型药敏试验为金标准,采用Xpert MTB/RIF系统行利福平耐药性检测的敏感度为93.33% (28/30),特异度为94.12% (32/34)。Xpert MTB/RIF系统平均检测耗时为2.1 h (1.8~2.6 h)。结论 Xpert MTB/RIF是一种简便、快速、准确,且能够同时对脊柱结核临床标本行结核分枝杆菌检测与利福平耐药性检测的分子检测技术,具有潜在的临床应用价值。     

Xpert MTB/RIF在脊柱结核诊断及利福平耐药检测中的应用价值

目的:探讨Xpert MTB/RIF技术在脊柱结核诊断及利福平耐药检测中的应用价值。方法:选取109例初步诊断为脊柱结核患者的脓液标本,分别行抗酸染色、BACTEC MGIT 960液体快速培养和Xpert MTB/RIF试验,对比3种检测结核分枝杆菌的敏感性及特异性的差异。对不同方法获取的脓液标本行Xpert MTB/RIF检测,评估脓液标本本身对Xpert MTB/RIF检测结核分枝杆菌效能的影响。以BACTEC MGIT 960液体快速培养药敏试验结果为金标准,分析Xpert MTB/RIF检测利福平耐药的效能。结果:抗酸染色、BACTEC MGIT 960液体快速培养及Xpert MTB/RIF检测的总体敏感性分别为25.92%、48.15%和77.78%。Xpert MTB/RIF检测开放手术、B超定位穿刺和穿刺活检取得脓液标本的敏感性分别为83.78%、76.47%和44.68%。以BACTEC MGIT 960液体快速培养药敏试验结果为金标准,Xpert MTB/RIF检测利福平耐药的敏感性和特异度分别为80%(4/5)和90.70%(39/43)。结论:Xpert MTB/RIF试验对脊柱结核具有较高的诊断价值,同时能对利福平耐药菌株进行检测,脓液标本中结核分枝杆菌含量对Xpert MTB/RIF检测的敏感性影响较大。     

Antituberculous therapy-induced fulminant hepatic failure: successful treatment with liver transplantation and nonstandard antituberculous therapy.

Standard antituberculous therapy including isoniazid, rifampin, ethambutol, and pyrazinamide is widely used for the treatment of active tuberculosis. Its most important side effect is hepatotoxicity, ranging from asymptomatic transaminitis to fulminant hepatic failure. A 19-year-old woman was admitted to our unit due to jaundice and unconsciousness. According to her past medical history, she was diagnosed as having extrapulmonary tuberculosis and had been prescribed standard antituberculous therapy. The patient became icteric and unconscious on the fourth day after therapy initiation. She was diagnosed with drug-induced acute fulminant hepatic failure and underwent living-related liver transplantation. Nonhepatotoxic antituberculous therapy (cycloserine, ciprofloxacin, streptomycin, and ethambutol) and low-dose immunosuppressive therapy were started after transplantation. Currently the patient is very well with normal graft function 42 months after transplantation. Here we report a case of a patient with acute fulminant hepatic failure caused by isoniazid, rifampicin, or both, who was successfully treated with living-related liver transplantation and a relatively less hepatotoxic antituberculous therapy. In conclusion, liver transplantation is a feasible therapy for individuals with standard antituberculous therapy-induced hepatic failure. Nonhepatotoxic antituberculous therapy may achieve control of active tuberculosis in such individuals after transplantation.     

Drug resistance patterns in 111 cases of drug-resistant tuberculosis spine

Purpose

We report the largest study conducted till date of drug resistant tuberculosis in spine analyzing the drug susceptibility patterns in 111 cases of proven drug resistance.

Methods

An observed cross-sectional study was conducted. Six-hundred and eighty-six patients with positive cultures underwent sensitivity testing to 13 commonly used anti-tubercular drugs using BACTEC MGIT-960 system.

Results

Females (60.3%) outnumbered males (39.6%). Only three patients (2.7%) were found HIV positive, and none of these had AIDS. Forty-four (39.6%) patients had taken AKT in the past for some form of tuberculosis. Eight (7.2%) patients had history of treatment default. The drug sensitivity testing revealed 87 (78.3%) cases of multi drug resistance (resistance to both isoniazid and rifampicin) and 3 (2.7%) cases of XDR-TB spine. Of the individual drugs, widespread resistance was present to both isoniazid (92.7%) and rifampicin (81.9%), followed by streptomycin (69.3%). Least resistance was found to kanamycin, amikacin and capreomycin.

Conclusion

It is recommended to do routine biopsy, culture and drug sensitivity testing in all patients of tuberculosis spine to guide selection of appropriate second-line drugs when required. In cases of non availability of drug susceptibility testing despite repeated attempts, it is suggested to use data from large series such as this to plan best empirical chemotherapy protocol.

     

Antibiotic resistant tuberculosis in the United Kingdom: 1993-1999

BACKGROUND: The re-emergence of tuberculosis as a global health problem over the past two decades, accompanied by an increase in tuberculosis drug resistance, prompted the development of a comprehensive national surveillance system for tuberculosis drug resistance in 1993. METHODS: The UK Mycobacterial Resistance Network (Mycobnet), which includes all mycobacterial reference and regional laboratories in the UK, collects a minimum dataset on all individuals from whom an initial isolate of Mycobacterium tuberculosis complex has been isolated and submitted by source hospital laboratories. Data sought include susceptibility to first line antibiotics, demographic, geographical, and risk factor information. RESULTS: There were 25 217 reports of initial isolates of M tuberculosis complex in the UK between 1993 and 1999. All were tested for sensitivity to isoniazid, rifampicin, and ethambutol and 12 692 of the isolates were also tested for sensitivity to pyrazinamide and streptomycin. A total of 1523 (6.1%) isolates were resistant to one or more drugs, 1397 isolates (5.6%) were resistant to isoniazid with or without resistance to other drugs, and 299 (1.2%) were multidrug resistant. Although the numbers of drug resistant isolates increased over the period, the proportions remained little changed. Certain groups of people were at a higher risk of acquiring drug resistant tuberculosis including younger men, residents of London, foreign born subjects, patients with a previous history of tuberculosis and those infected with HIV. CONCLUSION: Although the proportion of drug resistant tuberculosis cases appears to be stable in the UK at present, more than one in 20 patients has drug resistant disease at diagnosis and more than one in 100 has multidrug resistant disease. Tuberculosis control measures should be strengthened to minimise the emergence of drug resistance through rapid diagnosis, rapid identification of drug resistance, supervised treatment, and maintenance of comprehensive surveillance.     

Ultrashort-course chemotherapy for culture-negative pulmonary tuberculosis--a qualified success

A group of 250 patients with new or enlarging apical lung lesions which were thought to be tuberculous, and who had positive tuberculin tests but negative sputum smears and cultures for Mycobacterium tuberculosis, were treated with an ultrashort (3-month), 4-drug (rifampicin, isoniazid, pyrazinamide and ethambutol) regimen. One patient developed bacteriologically positive pulmonary tuberculosis (PTB) during the treatment period and 35 others (14%) developed bacteriologically positive PTB after completing the drug regimen.     

Bursectomy,Curettage, and Chemotherapy in Tuberculous Trochanteric Bursitis

We presented three patients with trochanteric tuberculosis and described the clinical and imaging findings of the infection. Histology revealed a necrotizing granulomatous bursitis and microbiology confirmed tuberculosis. All cases were successfully treated with bursectomy and curettage of the trochanteric lesion and antituberculous chemotherapy including isoniazid, pyrazinamide, rifampicin, and ethambutol.     

Short-course chemotherapy for pulmonary tuberculosis with a rifampicin-isoniazid-pyrazinamide combination tablet

The effectiveness of a tablet containing a combination of rifampicin, isoniazid and pyrazinamide (Rifater; Mer-National) in the treatment of pulmonary tuberculosis was examined by comparing it with a previously evaluated four-drug regimen. Of 150 black goldminers with a first case of pulmonary tuberculosis, 69 were randomly allocated to receive the combination tablet (RHZ), 5 tablets per day on weekdays for 100 treatment-days, and 81 the four-drug regimen (streptomycin, rifampicin, isoniazid and pyrazinamide) (RHZS). Non-compliance was detected in 42% of the RHZ group and in 16% of the RHZS group. Two patients in the RHZ group and 4 in the RHZS group had to have their treatment altered because routine investigations revealed drug-resistant mycobacteria. Treatment was unsuccessful in 10 patients in the RHZ group, with 4 men failing to complete the regimen and being lost to follow-up, 3 cases of failure of conversion of sputum on the regimen, and 3 relapses. The results for the RHZS group were similar, with 4 failures to complete the regimen, 2 treatment failures and 4 relapses. Evaluation of RHZ showed it to be comparable with a previously evaluated, successful short-course regimen (RHZS). The high incidence of non-compliance probably reflects reduced supervision of this wholly oral regimen.     

Pneumonitis induced by rifampicin

An 81-year-old man was admitted to hospital with pulmonary Mycobacterium tuberculosis infection and was treated with rifampicin (RFP), isoniazid (INH), and ethambutol (EB). On day 9 he developed fever and dyspnoea. Chest radiographs showed new infiltration shadows in the right lung. Bronchoalveolar lavage (BAL) was performed and increased numbers of lymphocytes were recovered. Drug induced pneumonitis was suspected so the antituberculous regimen was discontinued and methylprednisolone was administered. The symptoms and infiltration shadows improved. INH and EB were reintroduced without any recurrence of the abnormal shadows. T cell subsets in the BAL fluid and a positive lymphocyte stimulation test for RFP suggest that RFP induced pneumonitis may be related to a complex immunological response.     

Characteristics of patients with drug resistant and drug sensitive tuberculosis in East London between 1984 and 1992.

BACKGROUND--The aim of this study was to investigate retrospectively factors associated with drug resistant tuberculosis at the London Chest Hospital. METHODS--The microbiology results for patients with tuberculosis at the hospital for the period 1984-92 were reviewed, together with case notes and chest radiographs of all patients with drug resistant tuberculosis and of 101 patients with drug sensitive tuberculosis notified during the same period as a control group. RESULTS--Culture positive pulmonary tuberculosis occurred in 292 patients. Drug resistant strains were isolated from 20 patients (6.8%). Ten of the 292 (3.4%) had strains resistant to a single drug and nine (3.1%) had resistance to more than one first line drug. One patient had strains resistant to isoniazid and capreomycin. Strains resistant to more than one drug were all resistant to isoniazid and rifampicin. In five patients these strains were also resistant to pyrazinamide and in two they were resistant to streptomycin. Single drug resistant strains were resistant to isoniazid (nine patients) or streptomycin (one patient). Among the risk factors studied previous treatment for tuberculosis was the most significant association with drug resistant tuberculosis (7/9) for patients with resistance to more than one drug; 5/11 for single drug resistance compared with 6/101 patients in the drug sensitive group (odds ratio 22.8). Other risk factors were bilateral disease at presentation (odds ratio 8.5), and disease at a young age (odds ratio 1.03). CONCLUSIONS--Previous treatment for tuberculosis and bilateral disease at presentation were found to be more commonly associated with cases of drug resistant than with drug sensitive tuberculosis.