实时荧光定量PCR法分析结核分枝杆菌对异烟肼耐药的分析

目的利用实时荧光定量PCR技术快速检测异烟肼耐药结核分枝杆菌。方法收集到医院就诊的结核病疑似患者痰液样本,提取痰液样本的总DNA,利用实时荧光定量PCR(Real-time PCR)技术对结核分枝杆菌感染进行快速筛查,并与传统药敏试验进行比较,对两者的灵敏度、特异性、一致性进行比较分析。结果检测346例结核病人临床分离培养样本,药敏试验检出257例异烟肼敏感标本,101例异烟肼耐药标本;实时荧光定量PCR法共检测出异烟肼敏感和耐药标本225例98例,灵敏度为86.64%,特异性为93.92%,一致率为93.12%。结论跟传统药物敏感性实验相比,实时荧光定量PCR法检测速度快速、特异性强、灵敏度较高,可用于结核分枝杆菌耐异烟肼突变的快速检测,适于耐多药结核病的快速筛查。     

DNA微阵列芯片技术检测培阳肺结核患者痰样本中结核分枝杆菌耐药性的价值

目的 评价DNA微阵列芯片法(以下称芯片法)检测培养阳性患者痰样本的利福平和异烟肼相关耐药基因的效能.方法 收集并检测389例疑似肺结核患者痰样,以BACTEC MGIT 960液体药敏(以下称MGIT 960药敏)为参考标准,评价芯片法检测样本利福平(RIF)、异烟肼(INH)耐药性和MDR-TB的灵敏度、特异度及一...     

线性探针技术快速检测肺结核耐药性临床研究

目的评价线性探针技术快速检测涂阳肺结核耐药性的可靠性和及时性。方法随机抽取我院收治的120例涂阳肺结核患者痰标本,同时采用线性探针技术和传统培养药敏试验两种方法检测异烟肼和利福平耐药性,以传统培养药敏试验结果为金标准,分析线性探针技术诊断耐药结核病的灵敏性、特异性和及时性。结果线性探针技术检测异烟肼的灵敏性和特异性分别为100%、98.98%;检测利福平分别为96.15%、98.94%;对耐多药的检出率18.33%。24小时内可得到报告。结论线性探针技术能快速检测异烟肼、利福平的耐药性,有利于耐药肺结核特别是耐多药肺结核的快速筛查和临床及时诊治。     

Prevalence of resistance to anti-tuberculosis drugs: results of the 1998/99 national survey in Italy.

OBJECTIVE: To determine the prevalence of resistance to the main anti-tuberculosis drugs in newly and previously treated tuberculosis patients in Italy and to evaluate the contribution of foreign-born and human immunodeficiency virus (HIV) positive cases to drug resistance. METHODS: Methods and definitions were derived from the WHO/IUATLD Global Project on Anti-tuberculosis Drug Resistance Surveillance. Univariate and multivariate analysis was used to study prevalence rates of drug resistance in risk groups. RESULTS: In a national survey in Italy, 810 initial isolates of Mycobacterium tuberculosis (683 from new cases, 115 from retreatment cases and 12 from patients whose treatment history was unknown/dubious) were analysed. Low prevalence of drug and multidrug resistance was found in the new cases (isoniazid 2.9%; rifampicin 0.8%; multidrug resistance 1.2%; any drug resistance 12.3%). The prevalence of resistance to isoniazid and rifampicin was significantly higher in immigrants and HIV-positive subjects, respectively. A high prevalence of drug resistance was found in cases with previous treatment failure or default (isoniazid 5.2%; rifampicin 4.3%; multidrug resistance 36.5%; any drug resistance 61.7%). RECOMMENDATIONS: Special efforts are necessary to monitor trends in drug resistance and to ensure favourable treatment outcomes among immigrants and HIV-positive tuberculosis cases.     

134例老年肺结核患者痰结核菌耐药情况分析

目的探讨老年肺结核患者痰结核菌的耐药情况。方法采用ＢＡＣＴＥＣ法对１３４例老年痰菌阳性肺结核患者痰结核分支杆菌做耐药性检测。结果老年肺结核患者初始耐药率为３６８％，显著低于获得性耐药率的７５９％（Ｐ＜００１）；耐异烟肼（ＩＮＨ，Ｈ）、利福平（ＲＦＰ，Ｒ）、链霉素（ＳＭ，Ｓ）前者亦明显低于后者（Ｐ＜００１）。初治组耐２和３种药比例为６６％和５３％，与复治组的２２４％和２７７％相比差异有显著性（Ｐ＜００１）。至少耐异烟肼和利福平两种药的比例，初治组为９２％，显著低于复治组的５００％（Ｐ＜００１）。结论老年人肺结核耐药情况严重，尤其是获得性耐药和耐多药比例偏高，应引起足够重视     

Comparative study of bactec MGIT 960 AST and conventional proportion method using Ogawa medium for the drug susceptibility testing of Mycobacterium tuberculosis to isoniazid

OBJECTIVE: To evaluate the accuracy of drug susceptibility testing to isoniazid with BACTEC MGIT 960 (MGIT AST) comparing with the standard proportion method using Ogawa medium. METHOD: A total of 1109 M. tuberculosis strains, which were selected from the collection of RYOKEN drug resistance survey in 2002, were selected and subjected to the susceptibility testing to isoniazid using MGIT AST and 1% Ogawa standard methods. The results from MGIT AST were compared with the judicial diagnosis by Ogawa. The sensitivity to detect drug resistance, the specificity for susceptible strain, the efficiency of overall agreement, and kappa coefficient were calculated to evaluate the performance. The treatment process, outcome and prognosis were analysed for the patients on whom the tests showed discrepant results. RESULTS: Compared with the judicial results, the sensitivity, specificity, efficiency, and kappa coefficient of MGIT AST were 100%, 97.1%, 97.3%, and 0.798, respectively. The strains, which showed discrepant results between MGIT AST and Ogawa, were all susceptible by Ogawa and resistant by MGIT AST. A total of 11 out of 30 discrepant cases were followed clinically and no relapse cases were identified, irrespective of the modification of the treatment regimen. As for the proportion of primary INH drug resistance in the present study, it was 5.3% with MGIT AST but was 2.7% with Ogawa, and the difference was statistically significant (p = 0.005). DISCUSSION: The discrepancies on the results of drug susceptibility testing of M. tuberculosis strains to isoniazid between MGIT AST and 1% Ogawa proportion method have been reported. In the present study, the sensitivity, specificity, and overall efficiency of MGIT AST on the prevalent strains in Japan were all beyond 95%, and considered sufficient as the anti-tuberculosis drug susceptibility testing (AST), though 2.7% of discrepancy was observed. Even for the discrepant cases, there was no difference in the treatment outcome and prognosis. Thus, MGIT AST was confirmed as a reliable AST method comparable to Ogawa standard. However, MGIT AST might increase the proportion of INH resistance if it was used as a major AST method, compared with Ogawa.     

硝酸盐还原试验与反向斑点杂交试验联合检测结核分枝杆菌耐药性

目的 联合检测结核分枝杆菌耐药性,以提高其灵敏度和特异性.方法 对126株已知耐药浓度的结核分枝杆菌耐药株和敏感株分别采用硝酸盐还原试验( Nitrate Reductase Assay,NRA)和反向斑点杂交试验(reverse-dot blot hybridization,RBH),检测其异烟肼(INH)和对利福平(RFP)的耐药性,并将结果将与绝对浓度法结果相比较.结果 硝酸盐还原试验和反向斑点杂交试验联合检测与绝对浓度法比较有高度的一致性(P＞0.05),异烟肼灵敏度95.5％,特异性100％;利福平灵敏度100％,特异性98.1％.结论 硝酸盐还原试验和反向斑点杂交试验联合检测可作为快速结核分枝杆菌耐药性试验方法.     

山东省滨州市123例肺结核患者痰结核菌耐药情况分析

目的探讨肺结核患者痰结核菌的耐药情况。方法采用痰结核分支杆菌培养并做药物敏感性试验检测。结果肺结核患者耐药率为58.54%;耐异烟肼（INH,H）37.4%、利福平（RFP,R）13.8%、链霉素（SM,S）56.1%、乙胺丁醇（EMB,E）2.4%。至少耐异烟肼和利福平两种药的比例为16.26%。结论肺结核耐药情况严重,尤其是耐多药比例偏高,应引起足够重视。     

Study of drug resistance in previously treated tuberculosis patients in Gujarat, India.

SETTING: Department of Tuberculosis and Chest Diseases and State Tuberculosis Diagnosis and Training Centre (STDTC), a DOTS centre in Ahmedabad, Gujarat State, India. The study was carried out by retrospectively reviewing patient data between January 2000 and August 2001. OBJECTIVE: To evaluate the pattern of drug resistance among previously treated tuberculosis patients who remained symptomatic or smear-positive despite receiving anti-tuberculosis drugs under DOTS for a minimum of 5 months. DESIGN: A total of 1472 pulmonary tuberculosis patients who had taken anti-tuberculosis treatment were evaluated retrospectively with respect to their drug resistance pattern by sputum culture for acid-fast bacilli (AFB) and sensitivity testing with isoniazid, rifampicin, streptomycin and ethambutol (E). RESULT: Of the 1472 patients evaluated, 804 (54.6%) were treatment failure cases and 668 (45.4%) were relapse cases; 822 patients (373 failure and 449 relapse) were culture-positive. Of these 822 patients, 482 (58.64%, 261 failure and 221 relapse) were resistant to one or more drugs. Resistance to one drug was observed in 86 patients (10.46%), to two drugs in 149 (18.13%), to three drugs in 122 (14.84%) and to four drugs in 125 (15.21%). Single drug resistance was most commonly seen with isoniazid (62 patients, 7.5%), followed by streptomycin (12 patients, 1.4%), rifampicin (eight patients, 0.97%) and ethambutol (four patients, 0.4%). Resistance to isoniazid plus rifampicin alone was seen in 76 patients (9.2%). CONCLUSION: Drug resistance is a major problem in the treatment of pulmonary tuberculosis. Detection of drug resistance patterns and treatment with second-line anti-tuberculosis drugs in appropriate regimens are necessary in the treatment of failure and relapse cases in order to reduce the emergence of multidrug-resistant tuberculosis.     

Anti-tuberculosis drug resistance in the West Province of Cameroon.

SETTING: All 15 district hospitals of the West Province of Cameroon, between July 1997 and June 1998. OBJECTIVE: To determine the prevalences of initial and acquired resistance to the main anti-tuberculosis drugs 2 years after the implantation of a tuberculosis control programme in the province. METHODS: A total of 615 adults consecutively admitted to the 15 district hospitals with sputum smear-positive pulmonary tuberculosis were systematically studied. Sputum specimens collected from each patient were cultured on Lowenstein-Jensen medium. Testing of susceptibility to the major anti-tuberculosis drugs was performed by the indirect proportion method. RESULTS: Growth of Mycobacterium tuberculosis complex strains was obtained from specimens of 566 (92%) of the 615 patients. The overall resistance rate (one or more drugs) was 26.9%, with initial resistance being 19.7% (86/437) and acquired resistance 51.1% (66/129). Initial resistance to isoniazid was the most common (12.1%), followed by streptomycin (11.7%), ethambutol (2.5%) and rifampicin (2.1%). Initial resistance was noted as 13.5% to one drug, 4.3% to two, 1.1% to three and 0.7% to four. Acquired resistance to isoniazid was the most frequent (41.1%), followed by streptomycin (26.4%), rifampicin (14.7%) and ethambutol (9.3%). Acquired resistance was 25.6% to one drug, 14.7% to two, 7% to three and 3.9% to four. CONCLUSION: The proportion of resistant tuberculosis in the West Province is quite high. This underscores the need for the improvement of the control programme by introducing the DOTS strategy.     

MTBDRplus 2.0技术对疑似肺结核和耐多药结核病患者的诊断价值

目的评价二代线性探针(GenoType MTBDRplus VER 2.0,简称"MTBDRplus 2.0")技术快速检测疑似肺结核患者痰标本中结核分枝杆菌(MTB)及其对利福平和异烟肼耐药性的诊断价值。方法回顾性分析2018年1 12月就诊于西安市胸科医院的1065例疑似肺结核患者的实验室检测结果,剔除耐药不确定及痰标本污染者96例,最后纳入969例疑似患者作为研究对象。每例患者用同一份痰标本分别进行GeneXpert MTB/RIF(简称"GeneXpert")检测、MTBDRplus 2.0检测、BACTEC MGIT 960(简称"MGIT 960")液体培养,对培养阳性且鉴定为MTB的菌株进行MGIT 960药物敏感性试验(简称"MGIT 960药敏试验"),以MGIT 960液体培养结果为参考标准,评价MTBDRplus 2.0技术检测痰标本中MTB的检测效能;以MGIT 960药敏试验结果为参考标准,评价MTBDRplus 2.0技术检测利福平和异烟肼耐药性的检测效能。结果969例疑似肺结核患者中.MGIT 960液体培养检测MTB阳性409例,阴性560例。以MGIT 960液体培养检测结果为参考标准,MTBDRplus 2.0和GeneXpert检测疑似肺结核患者痰标本MTB的敏感度、特异度、Kappa值分别为91.0%(372/409)、93.8%(525/560)、0.848和92.9%(380/409)、92.5%(518/560)、0.850。以MGIT 960药敏试验结果为参考标准,MTBDRplus 2.0和GeneXpert检测利福平耐药性的敏感度、特异度、Kappa值分别为89.8%(53/59)、95.8%(295/308)、0.817和91.5%(54/59)、95.8%(295/308)、0.828;MTBDRplus 2.0检测异烟肼耐药性的敏感度、特异度和Kappa值分别为80.4%(82/102)、96.3%(260/270)和0.791。结论 MTBDRplus 2.0技术对检出疑似肺结核患者痰标本中的MTB,以及对利福平和异烟肼的耐药性均具有较好的检测效能。     

Anti-tuberculosis drug resistance in Portugal.

SETTING: A survey based upon a representative sample of smear-positive pulmonary tuberculosis patients was undertaken in Portugal, as part of the World Health Organization's Global Project on Anti-Tuberculosis Drug Resistance Surveillance. OBJECTIVE: To determine the level of primary antituberculosis drug resistance at both national and regional levels, and to assess its relative weight within the performance of the National Tuberculosis Programme (NTP). DESIGN: Mycobacterium tuberculosis isolates from 1,105 patients with smear-positive pulmonary tuberculosis admitted to 46 randomly stratified treatment centres all over mainland Portugal were submitted to susceptibility testing with four drugs. Human immunodeficiency virus (HIV) testing was included in the patients' evaluation scheme. RESULTS: Of the strains isolated, 197 (17.8%) were resistant to at least one drug. Primary resistance to isoniazid was 7.7% and to rifampicin 1.9%. Acquired drug resistance was 39.2% in total, any acquired resistance to isoniazid 31.1% and to rifampicin 20.9%. Primary multidrug resistance (MDR) was 1.8% and acquired MDR was 20.9%. HIV testing was positive in 29.2% of MDR-TB cases. CONCLUSIONS: Drug resistance in Portugal is high. Primary MDR and particularly acquired MDR occur in a high proportion of cases, indicating a need for improvement in NTP performance.     

Drug-resistant tuberculosis in Sierra Leone

Culture and sensitivity tests were performed on tubercle bacilli from patients with tuberculosis attending a provincial general hospital in Sierra Leone, who appeared clinically to have drug-resistant infections. Amongst the general intake of patients from 1978 to 1984 there was a 10.5% incidence of isoniazid resistance, with 7.7% of patients having strains resistant to both streptomycin and isoniazid, 1.3% resistant to rifampicin, and 0.8% resistant to ethambutol. Multiple drug resistance was more frequent than single drug resistance. This high incidence is largely due to inadequate and irregular supplies of first line drugs.     

Klinisch-experimentelle Untersuchungen zur Lungenfunktionsprüfung mit einem Thermistor als Atemstromrezeptor

(1) The dynamics of changes in drug resistance in tubercle bacilli were studied in 82 patients kept under observation over a lengthy period (up to 57 months). More than 30 cultures of tubercle bacilli were isolated from each of 18 patients during the trials and studied. (2) A difference was noted in the rate of establishment of bacterial resistance to three drugs (streptomycin, isoniazid, PAS) used in the treatment of tuberculosis patients. Under out-patient conditions resistance was most quickly acquired to streptomycin. Resistance to isoniazid was established considerably more slowly, while resistance to PAS took the longest to become established. (3) It was shown that during isoniazid treatment about 20 per cent of cultures isolated from patients after various periods of treatment remained sensitive to isoniazid. (4) Resistance to isoniazid reached a high level in the first seven months of treatment. There after only slight fluctuations occurred in the mean values for resistance to isoniazid in the bacilli populations. (5) It was noted that resistance to PAS occurs more frequently when a second course of treatment with that drug is given. Acquired resistance to PAS tends to be maintained for a long time after cessation of treatment. (6) The acquired resistance to streptomycin is maintained in most patients for a long period after cessation of streptomycin treatment; but in some cases it disappears quickly and completely. (7) For a long period, a study was maintained on correlations between drug resistance and catalase activity in tubercle bacilli isolated from the same patients. It was shown that catalase activity depends on the degree of isoniazid resistance and the range of drug sensitivity, which change during the treatment.     

Survey of drug resistance of Mycobacterium tuberculosis in 3 Mexican states, 1997

BACKGROUND: Drug resistance threatens global tuberculosis (TB) control efforts. Population-based estimates of drug resistance are needed to develop strategies for controlling drug-resistant TB in Mexico. OBJECTIVE: To obtain population-based data on Mycobacterium tuberculosis drug resistance in Mexico. METHODS: To obtain drug resistance data, we conducted a population-based study of TB cases in the states of Baja California, Sinaloa, and Oaxaca, Mexico. We performed cultures and drug susceptibility testing on M tuberculosis isolates from patients with newly diagnosed, smear-positive TB from April 1 to October 31, 1997. RESULTS: Mycobacterium tuberculosis was isolated from 460 (75%) of the 614 patients. Levels of resistance in new and retreatment TB cases to 1 or more of the 3 current first-line drugs used in Mexico (isoniazid, rifampin, and pyrazinamide) were 12.9% and 50.5%, respectively; the corresponding levels of multi-drug-resistant TB were 2.4% and 22.4%. Retreatment cases were significantly more likely than new cases to have isolates resistant to 1 or more of the 3 first-line drugs (relative risk [RR], 3.9; 95% confidence interval [CI], 2.8-5.5), to have isoniazid resistance (RR, 3.6; 95% CI, 2.5-5.2), and to have multi-drug-resistant TB (RR, 9.4; 95% CI, 4.3-20.2). CONCLUSIONS: This population-based study of M tuberculosis demonstrates moderately high levels of drug resistance. Important issues to consider in the national strategy to prevent M tuberculosis resistance in Mexico include consideration of the most appropriate initial therapy in patients with TB, the treatment of patients with multiple drug resistance, and surveillance or periodic surveys of resistance among new TB patients to monitor drug resistance trends.     

广州地区5年间耐多药结核分枝杆菌药物敏感试验结果分析

目的总结耐多药结核分枝杆菌药物敏感试验结果,为制定耐多药治疗方案提供依据。方法回顾分析过去5年广州市胸科医院收治的肺结核患者,其中痰液培养结核分枝杆菌药物敏感性试验结果为耐多药的菌株共605株,按年度和除异烟肼和利福平外的其他抗结核药物的种类进行统计,比较分析,分析各种抗结核药物的耐药率以及对比五年间耐药率的变化情况。结果阿米卡星的耐药率16.86%,克拉霉素的耐药率12.89%,对氨基水杨酸异烟肼的耐药率25.29%。乙胺丁醇的耐药率63.64%,链霉素的耐药率87.27%和利福布汀的耐药率73.06%。左氧氟沙星的耐药率41.15%,莫西沙星的耐药率40.49%,丙硫异烟胺的耐药率42.81%。结论在广州地区中,阿米卡星、克拉霉素和对氨基水杨酸异烟肼的耐药率比较低。     

基因芯片与线性探针技术检测涂阳肺结核患者痰标本MTB耐药性的价值

目的 评价基因芯片技术与线性探针技术(GenoType MTBDRplus)快速检测MTB耐药性的临床应用价值。方法 选取西安市胸科医院2017年4月至2018年8月住院治疗的493例涂阳肺结核患者作为研究对象,收集其痰标本,痰标本量均不少于2ml。每例患者用同一份痰标本分别进行基因芯片检测、线性探针检测和BACTEC MGIT 960液体培养(简称“MGIT 960液体培养”),同时对培养阳性且鉴定为MTB的临床分离株进行MGIT 960液体药物敏感性试验(简称“药敏试验”)。以MGIT 960液体药敏试验结果为参照标准,分析基因芯片技术及线性探针技术检测涂阳肺结核患者痰标本MTB利福平和异烟肼耐药性的效能。结果 454例研究对象同时具有3种药敏试验检测结果。以MGIT 960液体药敏试验结果为参照标准,基因芯片法和线性探针法检测涂阳肺结核患者痰标本MTB利福平耐药性的敏感度、特异度、Kappa值分别为89.0%(65/73)、96.1%(366/381)、0.82和90.4%(66/73)、96.1%(366/381)、0.83;检测涂阳肺结核患者痰标本MTB异烟肼耐药性的敏感度、特异度、Kappa值分别为80.2%(93/116)、96.7%(327/338)、0.80和81.9%(95/116)、97.0%(328/338)、0.82。454例涂阳肺结核患者痰标本中,453例应用2种分子生物学检测方法检测MTB利福平耐药性结果相同,符合率为99.8%;445例应用2种分子生物学方法检测MTB异烟肼耐药性结果相同,符合率为98.0%。结论 基因芯片技术和线性探针技术(GenoType MTBDRplus)检测涂阳肺结核患者痰标本MTB利福平和异烟肼耐药性与MGIT 960液体药敏试验具有较高的一致性,二者均可为临床提供快速、特异的耐药检测结果。     

Primary antituberculosis drug resistance and acquired rifampicin resistance in Gujarat, India

The prevalence of primary antituberculosis drug resistance in Gujarat, as studied between 1983 and 1986, was found to be significantly high, especially for isoniazid (13.9%) and streptomycin (7.4%). Primary rifampicin and pyrazinamide resistance were not detected in any strain. The prevalence of rifampicin resistance among treatment failure and relapse cases of pulmonary tuberculosis increased significantly from 2.8% in 1980 to 37.3% in 1986. In about 95% of the rifampicin resistant strains there was also resistance to isoniazid or streptomycin or both: resistance to isoniazid was detected in more than 90%.     

2006～2015年上海市公共卫生临床中心艾滋病患者的分枝杆菌培养阳性结果分析

目的 分析2006年～2015年上海市公共卫生临床中心病原培养分枝杆菌阳性的艾滋病患者结核分枝杆菌（MTB）与非结核分枝杆菌（NTM）的构成及耐药情况。方法 收集2006年1月～2015年12月入住我院且存在临床相应感染症状并具有分枝杆菌培养阳性结果的艾滋病患者816例,收集其年龄、性别等一般资料、耐药检测和NTM分型结果并进行分析。结果 816例分枝杆菌感染患者中男679例,女137例,中位年龄40岁。MTB感染患者364例（44.6%）,NTM感染患者377例（46.2%）。判定为MTB感染且有耐药检测资料患者共275例,对链霉素及异烟肼的耐药率均为24.0%（66例）,对乙胺丁醇的耐药率为17.1%（47例）,对利福平的耐药率为16.7%（46例）;耐多药结核病为42例（15.3%）。判定为NTM感染且有耐药检测资料患者共124例,对异烟肼的耐药率为91.1%（113例）,对链霉素的耐药率为83.9%（104例）,对利福平的耐药率为47.6%（59例）,对乙胺丁醇的耐药率38.7%（48例）。共102例NTM患者有分型资料,其中鸟-胞内分枝杆菌33例（32.3%）,戈登分枝杆菌29例（28.4%）,堪萨斯分枝杆菌25例（24.5%）。结论 对于分枝杆菌培养结果阳性的艾滋病患者,NTM比例明显高于普通人群,治疗上需根据药物敏感试验结果适当调整用药。