BACTEC MGIT 960用于结核分枝杆菌直接药敏试验的研究

目的探讨BACTEC MGIT 960系统在结核分枝杆菌直接药敏试验的临床应用价值。方法对200株临床涂阳痰标本应用BACTEC MGIT 960系统进行一线抗结核药（链霉素、异烟肼、利福平、乙胺丁醇）结核分枝杆菌直接药敏试验,并与同步进行的BACTEC MGIT 960间接药敏试验及传统改良罗氏（L-J）比例法药敏试验进行比较,从结果报告时间和符合率等方面进行评价。统计学分析分别采用配对资料的t检验和Kappa检验。结果 BACTEC MGIT960直接药敏试验报告时间为（9.7±3.1）d,与BACTEC MGIT 960间接药敏试验报告时间（17.5±6.6）d比较差异有统计学意义（t=14.7,P〈0.01）,与L-J法报告时间（50.4±8.6）d比较差异有统计学意义（t=73.4,P〈0.01）;4种一线抗结核药物链霉素、异烟肼、利福平、乙胺丁醇药敏BACTEC MGIT 960直接法与BACTEC MGIT 960间接法药敏结果总符合率为97.3%,BACTEC MGIT 960直接法与L-J法药敏结果总符合率为96.2%。结论 BACTEC MGIT 960直接药敏试验快速、准确,可用于结核分枝杆菌的药物敏感性检测,利于耐药结核病的早期诊断和治疗。     

培养管变色硅胶显色法快速检测结核菌耐药性

目的评价一种快速检测结核分枝杆菌(Mycobacterium tuberculosis,简称MTB)耐药性的可视化变色硅胶培养管的性能,并进行方法学建立。方法以BACTEC MGIT960药敏结果作为标准,应用变色硅胶培养管检测50株MTB临床分离株对利福平(Rifampicin,简称RFP)、异烟肼(Isoniazid,简称INH)两种一线抗结核药物的耐药性,并对最佳检测条件进行探索;其次对100份结核病患者涂阳痰标本用变色硅胶培养管进行培养及药敏检测,结果与MGIT960药敏结果作比较,判断灵敏度、特异度、准确度。结果培养管变色硅胶显色法检测50株MTB临床分离株对RFP、INH药敏结果与BACTEC MGIT960符合率均达到100%,最佳检测时间为7-10d;培养管变色硅胶显色法检测100份涂阳痰标本对RFP药敏结果的灵敏度、特异度、准确度分别为87.5%、98.6%、95.8%;对INH药敏结果的灵敏度、特异度、准确度分别为86.6%、96.9%、93.8%,变色硅胶显色法检测2种抗结核药物结果与MGIT960法比较无差异(RFPχ~2=0.50,P0.05;INHχ~2=0.75,P0.05)。最佳检测时间为15-20d。结论培养管变色硅胶显色法检测MTB的耐药性具有成本低、快捷、操作简便、便于观察判断等优点,为检测MTB耐药提供了一种较好的方法,适合推广使用。     

70例MDR-TB病例抗结核二线药物敏感性检测结果分析

了解耐多药结核分枝杆菌临床分离株对二线药物耐药情况,评价BACTEC MGIT960系统在抗结核二线药敏试验中的临床应用价值。方法 收集本院的痰培养阳性、经菌种鉴定和一线药物的药物敏感性试验确定为耐多药的结核分枝杆菌菌株。采用BACTEC MGIT960对4种二线药氧氟沙星（OFLX）、左氧氟沙星（LVFX）、阿米卡星（AMK）和卷曲霉素（CPM）进行药物敏感性检测,并与改良 L-J罗氏培养法相比较。结果 (1)2008年10月-2009年4月,70例MDR-TB分离到的菌株,包括初治病例26例、复治病例44例,对四种药的总耐药率为61.4%（43/70）。对OFLX、LVFX、AMK和CPM耐药率分别为48.6%、40.0%、17.1%、7.1%。初治耐药率为57.7%（15/26）,复治耐药率达63.6%（28/44）。符合XDR-TB诊断者5例（7.1%）,包括初治者1例（1.4%）,复治者4例（5.7%）。（2）BACTEC MGIT960二线药物敏感试验与改良L-J法相比,平均报告时间仅10.2天,明显短于L-J法,符合率在90%以上。结论 （1） 在住院耐多药结核病患者中对这四种抗结核二线药物的耐药率已经到了一个较高水平;同时,当地存在XDR-TB,应该引起重视。（2）BACTEC MGIT960系统可以准确而快速地检测结核分枝杆菌对二线药物的耐受性.     

MGIT 960系统行二线抗结核药物敏感性试验的实用性评价

目的 评价全自动BACTEC MGIT 960系统在结核分枝杆菌二线抗结核药物（second-line anti-TB drugs,SLD）药敏试验中的实用性。 方法 本实验纳入患者435例。该纳入患者均通过痰涂片抗酸染色（萋-尼法）镜检,L-J固体和MGIT 960系统液体培养及鉴定,共筛查出阳性菌株212例。212例阳性菌株分别进行一线抗结核药物异烟肼（INH）、利福平（RFP）、链霉素（S）和乙胺丁醇（EMB）的耐药性试验,其中耐药菌株（所有耐药株）118例。以118例耐药株作为SLD药敏试验实验株,应用L-J和MGIT 960两种方法分别进行卷曲霉素（Cm）、卡那霉素（Km）、氧氟沙星（Ofx）和乙硫异烟胺（Eto）的药物敏感性试验,并计算出两方法药敏试验所需的时间和符合率;L-J和MGIT960两种培养方法的比较采用配对设计的卡方检验;两种方法检测结果的一致性用Kappa值检验判别。 结果 MGIT 960系统药敏试验所需时间平均10.0 d（1183/118）,L-J固体培养法所需时间平均28.5 d（3360/118）,两者相差18.5 d。MGIT 960与L-J培养法的SLD Cm、Km、Ofx和Eto敏感及耐药符合率和一致性检验Kappa值分别为88.1%（104/118）和0.520;95.8%（113/118）和0.815;91.5%（108/118）和0.824;71.2%（84/118）和0.375。 结论 MGIT 960系统液体培养法常用SLD药敏试验与金标准（L-J培养）比较除Eto外符合率高、一致性好;该方法检测快速、准确,具有很强的实用性,可作为结核分枝杆菌快速培养和药敏试验的新方法。     

耐药基因芯片对初治菌阳肺结核治疗指导价值研究

目的研究结核分枝杆菌(Mtb)耐药基因芯片法检测对初治菌阳肺结核治疗指导价值。方法应用Mtb耐药基因芯片检测100例初治肺结核感染菌株对利福平(RFP)、异烟肼(INH)、链霉素(SM)、乙胺丁醇(EMB)的耐药性,并与BACTEC MGIT960系统检测结果比对,经鉴别罗氏培养基(PNB)除外非结核分枝杆菌(NTM)结果 Mtb耐药基因芯片法检测耐RFP、INH、SM、EMB的敏感性和特异性分别为64.29%、75.00%、52.94%、62.5.%;93.02%、82.5%、93.98%、97.83%。结论结核分枝杆菌耐药基因芯片对于5种基因相关突变所致耐药性的敏感性略低,特异性较高,对初治菌阳肺结核临床治疗有一定参考价值。     

589株结核分枝杆菌对10种药物的耐药性分析

目的了解深圳市第三人民医院结核病患者结核分枝杆菌对10种药物的耐药情况。方法用MB/Bact 240分枝杆菌培养仪和改良罗氏管对患者的多种标本进行分枝杆菌分离培养鉴定,对分离到的结核分枝杆菌采用绝对浓度法对10种抗结核药物,利福平（RFP）、异烟肼（INH）、乙胺丁醇（EMB）、链霉素（SM）,利福喷丁（RFT）、丙硫异烟肼（TH）、氧氟沙星（OFL）、卷曲霉素（CPM）、卡那霉素（KM）和对氨基水扬酸（PAS）进行药敏试验。结果 1722例患者的标本结核分枝杆菌培养阳性589株,阳性率为34.2%（589/1722）,589株结核分枝杆菌对10种药总耐药率38%（224/589）,单耐药率最高为SM（17.1%）,最低为PAS（3.4%）;55株MDR-TB对另外8种药物耐药率最高为RFT（94.5%）,最低为KM（20%）和PAS（20%）,对10种药物全耐者有3株（0.5%）。结论结核菌耐药情况仍然十分严重,应加强抗结核药物的耐药性监测;根据药敏试验结果选择科学有效的化疗方案。     

骨关节结核各类标本进行结核分枝杆菌培养与PCR检测的阳性率结果分析

目的 比较骨关节结核病灶中脓液、干酪、肉芽及死骨组织的结核分枝杆菌培养阳性率,及结核分枝杆菌BACTEC MGIT 960（简称“MGIT 960”）培养、改良罗氏培养基培养、PCR检测的联合阳性率.方法 2011年6月至2012年5月首都医科大学附属北京胸科医院收治的经病理明确诊断为骨关节结核的患者50例,通过手术获取病灶中的脓液（50份）、干酪（42份）、肉芽（48份）及死骨组织标本（43份）,分别进行结核分枝杆菌MGIT 960培养及改良罗氏培养基培养,比较4种组织标本的培养阳性率,计算2种培养方法的联合阳性率;对脓液标本单独通过PCR方法进行了阳性率检测;计算脓液标本3种方法的联合阳性率.结果 4种标本MGIT 960培养的阳性率依次为：死骨组织（41.9％,18/43）＞脓液（40.0％,20/50）＞肉芽（33.3％,16/48）=干酪（33.3 ％,14/42）;4种标本改良罗氏培养阳性率依次为：肉芽（20.8％,10/48）＞脓液（20.0％,10/50）＞死骨组织（16.3 ％,7/43）＞干酪（14.3％,6/42）;脓液的PCR检测阳性率为48.0％（24/50）.MGIT 960和改良罗氏培养2种结核分枝杆菌培养方法的联合阳性率为50.0％（25/50）,而PCR检测、MGIT960和改良罗氏培养3种方法的联合阳性率为68.0（34/50）.结论 骨关节结核患者,应尽可能选择多种标本以提高培养阳性率.MGIT 960培养可以作为首选的细菌学诊断方法,但如果再联合使用改良罗氏培养和PCR扩增技术,能够进一步提高检出阳性率.     

MGIT960与罗氏培养法在结核分枝杆菌培养及药敏试验中的比对分析

目的 比对全自动Bactec MGIT960（MGIT960）系统与改良罗氏（L-J）培养方法 2者在结核分枝杆菌培养、鉴定及药敏试验中的优越性。 方法 按国家疾病预防控制中心（CDC）-碧迪项目辽宁工作方案要求和国际标准WS288-2009《肺结核诊断标准》纳入临床病例。本实验共纳入病例435例,其中男性319例,女性116例,平均年龄52.3岁（10～86岁）。上述每份病例标本均分别进行痰液涂片抗酸染色（萋-尼法）镜检,L-J固体培养和MGIT960系统液体培养。培养阳性者分别进行一线药物异烟肼（INH）、利福平（RIF）、链霉素（STR）和乙胺丁醇（EMB）的L-J法药敏试验（比例法）和MGIT960药敏试验;分别计算出抗酸染色镜检与MGIT960和L-J培养的符合率; L-J培养和MGIT960培养阳性检出率;L-J和MGIT960两种培养方法药敏试验所需的培养时间及符合率等。 结果MGIT960培养法和L-J培养法与痰涂片镜检的阳性符合率分别为90.8%（187/206）和89.3%（184/206）;MGIT960与L-J结果符合率92.8%（219/236）;MGIT960和L-J平均培养时间分别为9.5 d和31.5 d;MGIT960和L-J一线药物INH、RIF、STR和EMB敏感性试验的符合率分别为92.7%、95.9%、91.8%和97.3%。 结论 全自动MGIT960系统培养方法的敏感度和特异度高于L-J培养法,报告时间平均缩短至9.5-d,该系统是目前最为理想的快速结核分枝杆菌培养、鉴定和药敏试验检测系统。     

三种实验室技术在涂阳患者中诊断肺结核及其耐药性的效能分析

目的 探讨BACTEC MGIT 960分枝杆菌液体培养(简称“MGIT 960培养”)、GeneXpert MTB/RIF(简称“GeneXpert”)和荧光PCR熔解曲线技术在涂阳肺结核患者中的检测效能和应用意义。方法 回顾性分析2016年10月至2018年2月沈阳市胸科医院收治的所有涂片阳性患者411例,经结核分枝杆菌抗原检测(MPB64检测)阳性并符合临床诊断标准,确诊肺结核患者392例,MPB64检测阴性且经基因芯片菌种鉴定为非结核分枝杆菌感染患者19例,比较MGIT 960培养、GeneXpert和荧光PCR熔解曲线技术对涂阳肺结核患者的检测效能;以比例法药物敏感性试验(简称“药敏试验”)为参考标准,分别比较三种方法对耐药表型和耐药基因的检测效能。结果 在临床确诊的392例涂阳肺结核患者中,MGIT 960培养、GeneXpert和荧光PCR熔解曲线技术对痰标本检测的敏感度分别为96.17%(377/392)、99.74%(391/392)和92.09%(361/392),特异度均为100.00%(19/19)。以比例法药敏试验为参考标准,GeneXpert和荧光PCR熔解曲线技术对利福平(RFP)耐药检测的敏感度相同,均为94.44%(119/126);荧光PCR熔解曲线技术在异烟肼(INH)、左氧氟沙星(Lfx)和阿米卡星(Am)耐药检测上的敏感度分别为74.84%(116/155)、83.33%(75/90)、44.00%(22/50);MGIT 960培养对RFP、INH、Lfx和Am耐药检测的敏感度分别为96.03%(121/126)、98.06%(152/155)、100.00%(50/50)、96.67%(87/90)。结论 GeneXpert法简便、快捷、安全,能够快速诊断肺结核及检测RFP是否耐药。荧光PCR熔解曲线技术可检测4种药物耐药情况,检测时间短,临床快速诊断耐多药肺结核/广泛耐药结核病,对指导临床用药具有重要意义。MGIT 960培养与比例法药敏试验耐药检测的一致性极高,是目前快速诊断结核病及一线药耐药检测的常用方法,可以缩短表型药敏试验时间。     

BACTEC-MGIT960快速培养药敏对肺结核诊治的应用和评价

目的 BACTEC—MGIT960快速培养药敏在肺结核诊治中的应用评价。方法 采用BACTEC—MGIT960进行快速培养药敏，并与改良罗氏培养基和改良7H9培养基进行比较。结果 BACTEC—MGIT960培养报告时间仅8．5d，明显短于改良7H9培养基(20．6d)和改良罗氏培养基(30．8d)。BACTEC—MGIT960药物敏感试验结果与其它检测方法相比，符合率在90％以上，平均报告时间仅6．5d，明显短于改良7H9培养基(17d)和改良罗氏培养基(26．5d)。结论 BACTEC—MGIT960快速培养药敏有助于肺结核的诊断和治疗，是一种快速、有效、可靠的方法。     

BACTEC MGIT-960临床应用与结果分析

目的探讨BACTEC MGIT-960全自动分枝杆菌检测技术在结核病中的应用价值。方法用BACTEC MGIT-960对不同种类的标本进行分离培养及药敏试验．并与改良罗氏培养基法对比。结果BACTEC MGIT-960全自动分枝杆菌检测技术初代分离阳性时问平均为8．9天．药敏平均时间为7．1天。2152份各类标本总阳性率为29．23％．其中痰标本阳性率为31．22％。617株结核分枝杆菌的总耐药率为52．19％，其中初始耐药率为41．77％，获得性耐药率为74．24％，耐多药率(MDR)为17．02％。结论BACTEC MGIT-960全自动分枝杆菌检测技术是目前一种比较理想的快速检测结核菌的方法。     

Time to detection of Mycobacterium tuberculosis in BACTEC systems as a viable alternative to colony counting

OBJECTIVE: To investigate whether time to detection (TTD) of Mycobacterium tuberculosis in BACTEC Mycobacteria Growth Indicator Tube (MGIT) 960 and BACTEC 460 TB systems can be used as an alternative to colony-forming unit (cfu) counting. DESIGN: A single sputum sample recovered from each of 22 patients with tuberculosis (TB) was cultured on Middlebrook 7H11 agar and in BACTEC MGIT 960 and BACTEC 460 to investigate the relationship between cfu/ml and TTD. The relationship between TTD and treatment response was investigated by culturing a single sputum sample from each of 125 patients with TB in the BACTEC 460 system and comparing TTD values with their treatment response. RESULTS: An inverse correlation between TTD and bacterial number, as assessed by inoculum size and cfu/ml, was observed. For the 125 patients followed up during treatment, TTD values at diagnosis correlated with smear conversion rates at 2 months and treatment outcomes. Drug resistance of the infecting strain was associated with decreased killing, as indicated by the delayed increase in TTD during the first few days of treatment. CONCLUSION: The TTD of M. tuberculosis in BACTEC MGIT 960 and BACTEC 460 TB systems is a viable alternative to colony counting. TTD in liquid culture will facilitate mycobacterial quantification, especially in the evaluation of early bactericidal activity.     

Should chemotherapy for pulmonary tuberculosis be started after the completion of pretreatment sputum examination?

PURPOSE: Investigation on the influence of early start of chemotherapy on the results of sputum smear and culture (Ogawa medium and BACTEC MGIT960). OBJECT AND METHOD: Retrospective study for tubercle bacilli positive pulmonary tuberculosis cases in our hospital. RESULT: Starting chemotherapy before the completion of pretreatment sputum examination was found to have no adverse effect on smear positive rate and smear positive grade (246 cases), culture contamination rate on Ogawa medium and BACTEC MGIT960 (1128 sputums), culture positive rate and culture positive grade/time to confirm positive on Ogawa medium (245 cases), and culture positive rate on BACTEC MGIT960 (239 cases). Time to confirm positive on BACTEC MGIT960 was slightly, but significantly prolonged (1.16 days delay in cases with 1-3 days chemotherapy [228 cases], and 4.01 days delay in cases with 4-6 days chemotherapy [59 cases]). CONCLUSION: Unfluence of early start of chemotherapy on the results of sputum smear and culture is minor. In most of bacilli positive pulmonary tuberculosis cases chemotherapy can be started before the completion of pretreatment sputum examination.     

Characterization of mycobacterium isolates from pulmomary tuberculosis suspected cases visiting Tuberculosis Reference Laboratory at Ethiopian Health and Nutrition Research Institute,Addis Ababa Ethiopia:a cross sectional study

Objective:To characterize mycobaclerium isolates from pulmomary tuberculosis suspected cases visiting National Tuberculosis Reference Laboratory at Ethiopian Health and Nutrition Research Institute,for diagnosis of pulmonary tuberculosis from January 4 to February 22.2010 with total samples of 263.Methods:Sputum specimens were collected and processed:the deposits were cultured.Por culturing Lowenstein Jensen medium(LJ) and Mycobacteria Growth Indicator Tube(BACTEC MGIT 960) were used.Capilia Neo was used for detecting NTM isolates from isolates of BACTEC MGIT960.In Armauer Hansen Research Institute,Addis Ababa Ethiopia,Deletion typing PCR method for species identification(from confirmed Mycobacterium tuberculosis complex(MTBC) isolates by Capilia Neo) uas done.Results:Out of 263 enrolled in the study.124 and 117 ol them were positive for mycobaeterium growth by BACTEC MGIT 960 and 1.1 culture method,respectively.From BACTEC MGIT 960 positive media of 124 isolates.117 were randomly taken to perform Capilia TB Neo lest.From these 7(6%) of them were found to be NTM and 110(94%) were MTBC.From these 110 MTBC isolates,81 of them were randomly taken and run by the deletion typing RD9 PCR method of molecular technique.Out of these 78(96.3%) were found to be species of Mycobacterium tuberculosis and 3(3.7%) were found to be not in the MTBC.Regarding the types of methods of culture media.Mycobacteria Growth Indicator Tube(BACTEC MGIT 960) method was found to have excellent agreement(with kappa value ol 0.78) with the routine method of LJ.Conclusions:Pulmonary tuberculosis suspected cases visiting the National Tuberculosis Reference Laboratory at EHNR1 that were confirmed to be pulmonary tuberculosis are caused by the species of Mycobacterium tuberculosis.hence treatment regimen including pyrazinamide can be applied to the patients as the first choice in the study area in Addis Ababa.Ethiopia.There is indication of the presence of NTM in patients visiting the tuberculosis reference laboratory and this is important because NTM is known lo cause pulmonary disease similar with sign and symptom ol pulmonary tuberculosis but different in treatment.BACTEC MGIT 960 has excellent agreement with LJ media but it has high tendency of having high contamination rale unless a better decontamination method is designed.     

Comparison of the BACTEC MGIT 960 with L?wenstein-Jensen medium for recovery of mycobacteria from clinical specimens.

SETTING: Taiwan Provincial Chronic Disease Control Bureau. OBJECTIVE: To evaluate the rate of recovery and the mean time to detection (TTD) of mycobacteria in clinical specimens with two culture systems, the BACTEC MGIT 960 and L?wenstein-Jensen (LJ) medium. DESIGN: We studied 365 specimens, collected from 166 patients. Specimens were processed with standard N-acetyl-L-cysteine (NALC)-NaOH method, then inoculated onto BACTEC MGIT 960 and onto LJ slants. RESULTS: A total of 124 mycobacterial isolates (114 Mycobacterium tuberculosis and 10 non-tuberculous mycobacteria) were detected. The recovery rates were 94% (117/124) with BACTEC MGIT 960 and 75.8% (94/124) with LJ. The rates of contamination for each of the systems were 5.5% with BACTEC MGIT 960 and 4.1% with LJ. The TTDs for mycobacteria were 10.7 days with BACTEC MGIT 960 and 30.6 days with LJ. Excluding the non-tuberculous mycobacteria, the TTDs for M. tuberculosis were 11.1 days with BACTEC MGIT 960 and 30.7 days with LJ. The difference in TTD between smear-positive and smear-negative specimens for either mycobacteria (10.0 vs 12.6 days; P = 0.06) or M. tuberculosis (10.1 vs 12.7 days; P = 0.06) with BACTEC MGIT 960 was not statistically significant. CONCLUSION: The BACTEC MGIT 960 system can expedite the recovery of mycobacteria in culture. Combined with conventional solid medium, it also increases the overall recovery of mycobacteria in culture.     

Recovery rates of mycobacterium from suspected extra-pulmonary tuberculosis patients using liquid culture at a tertiary referral centre of India

Tuberculosis still remains a serious public health problem in developing countries. Rapid isolation of mycobacteria is critical for accurate diagnosis and management of tuberculosis. In the present study BACTEC MGIT 960 system was evaluated against Lowenstein Jensen (LJ) medium for isolation of mycobacteria from different extra-pulmonary specimens (N = 371). The samples were processed using NaOH-NALC method and inoculated in BACTEC MGIT and on LJ medium. The BACTEC MGIT 960 system detected 93 (25.06%) samples positive for acid fast bacilli and by LJ only 38 samples (10.24%) was positive. Furthermore, total 99 (26.68%) samples were detected positive by both the culture methods. The mean turnaround time to detection of mycobacteria by MGIT 960 were significantly less (12.4 days) as compared with LJ (22.76 days). In conclusion, BACTEC MGIT 960 system is more sensitive and rapid culture system for isolation of mycobacteria. However LJ culture method also suggested to further increase the detection rate of EPTB cases.