Xpert MTB/RIF系统在盆腔结核诊断中的应用价值分析

目的探讨Xpert MTB/RIF系统检测盆腔积液中结核分枝杆菌对于盆腔结核的诊断价值。方法采集30例临床确诊为盆腔结核患者及30例盆腔恶性肿瘤患者(对照组)的盆腔积液,对积液同时采用离心涂片法、荧光定量PCR法和Xpert MTB/RIF系统进行结核分枝杆菌检测,比较三种方法的敏感性及特异性。结果离心涂片法、荧光定量PCR法和Xpert MTB/RIF系统检测结核分枝杆菌的敏感性分别为6.67%(2/30)、53.33%(16/30)和73.33%(22/30),三种方法间差异均有统计学意义(P均<0.05)。三种方法特异度分别为100.0%,96.7%,100.0%,差异无统计学意义。结论采用Xpert MTB/RIF系统对盆腔积液进行结核分枝杆菌检测敏感性高,特异度好,具有较高的临床应用价值。     

全自动半巢式实时荧光定量PCR技术检测 BALF诊断肺结核的价值

目的 探讨全自动半巢式实时荧光定量PCR技术（GeneXpert MTB/RIF）检测肺胞灌洗液（BALF）诊断肺结核的价值。方法 采集2016年6月~2018年6月滨州市结核病防治院已经确诊的住院菌阴肺结核269例设为实验组,另收集其他肺部疾病患者87例设为对照组。利用支气管镜术收集肺胞灌洗液;肺胞灌洗液采用涂片法、培养法和GeneXpert MTB/RIF法3种方法检测结核分枝杆菌,分别对其阳性率及检测菌量进行比较。结果 BALF经涂片、培养和GeneXpert MTB/RIF3种方法检测,敏感度分别为23.05%、33.83%和62.08%,特异度分别为97.70%、97.70%和100.00%。GeneXpert MTB/RIF法分别与涂片法和培养法结果比较,差异均有统计学意义（P＜0.05）;涂片法与培养法结果比较,差异有统计学意义（P＜0.05）;对照组涂片和培养均出现2例阳性者,而GeneXpert MTB/RIF为阴性;涂片和培养相同的2例阳性者,经基因芯片检测确定为NTM。269例菌阴肺结核BALF涂片阴性者207例,207例涂片阴性者经GeneXpert MTB/RIF法检测呈阳性者105例,其中GenXpert MTB/RIF 法“极低”量级50例,占47.62%。269例菌阴肺结核BALF培养阴性者178例,178例涂片阴性者经GeneXpert MTB/RIF法检测呈阳性者76例,其中GenXpert MTB/RIF 法“极低”量级37例,占48.68%。结论 GeneXpert MTB/RIF技术结核分枝杆菌阳性检出率高于涂片和培养法,对早期诊断肺结核具有一定的临床应用价值,且操作简单、快速,值得推广。     

Xpert Mtb/RIF检测与BACTEC MGIT960结核培养药敏联合检测在结核病诊治中的应用分析

目的评估Xpert Mtb/RIF检测与BACTEC MGIT 960结核培养药敏联合检测在结核病诊治中的应用价值。方法对临床结核病患者的标本进行Xpert Mtb/RIF检测,同时进行BACTEC MGIT 960结核培养、鉴定、药敏试验。以BACTEC MGIT 960结核培养及药敏试验为金标准,分析Xpert Mtb/RIF检测的敏感性和特异性。结果以BACTEC MGIT 960结核培养及药敏试验为金标准,Xpert Mtb/RIF检测结核分枝杆菌的敏感性为87.7%(100/114),Xpert Mtb/RIF检测利福平耐药的敏感性为90%(45/50)、特异性为91.3%(53/58)。结论 Xpert Mtb/RIF检测敏感性及特异性高, BACTEC MGIT 960结核培养及药敏试验联合应用有助于及早进行有效诊治。     

不同荧光定量PCR试剂盒在结核分枝杆菌检测中的应用

目的比较两种不同荧光定量PCR试剂盒在福尔马林固定石蜡包埋(formalin fixed and paraffin embedded,FFPE)组织结核分枝杆菌(mycobacterium tuberculosis,MTB)核酸检测中的应用。方法选取40例经临床确诊结核病患者的FFPE组织标本,运用凯杰MTB核酸PCR检测试剂盒、晶芯分枝杆菌核酸PCR检测试剂盒及抗酸染色法进行检测,分析不同方法的MTB检出率。结果凯杰PCR法、晶芯PCR法及抗酸染色法的MTB检出率分别为80%(32/40)、37. 5%(15/40)、50%(20/40),各组之间差异有统计学意义(P 0. 01);凯杰PCR法的MTB检出率明显高于晶芯PCR法和抗酸染色法(P 0. 01)。结论凯杰PCR试剂盒检测FFPE组织中MTB的灵敏度高于晶芯PCR试剂盒,但晶芯PCR试剂盒能够同时检测并区分MTB和非结核分枝杆菌。     

Xpert结核分枝杆菌/利福平试验对结核病及耐多药结核病诊断价值的Meta分析

目的 评估Xpert结核分枝杆菌/利福平（MTB/RIF）试验对结核病的诊断价值.方法 检索PubMed、Medline、中国知网、万方数据库等,收集Xpert MTB/RIF试验对结核病诊断价值的文献,检索起止时间均为建库至2012年6月.2名研究者独立进行资料提取和文献质量评估.采用Meta-Disc 1.4软件进行Meta分析.结果 共纳入26篇文献,其中2篇文献涉及儿童病例,包含了13 270例来自临床患者的检测标本.Meta分析结果显示,Xpert MTB/RIF试验诊断结核病的汇总敏感度为87%（95%CI：86%～88%）、特异度为97%（95%CI：97%～97%）.按照结核病的类型和患者年龄进行亚组分析,Xpert MTB/RIF试验诊断肺结核的敏感度高于肺外结核病,90%（95%CI：89%～91%） vs 76%（95%CI：72%～79%）;诊断涂阴菌阳性和涂阳菌阳性结核病的敏感度分别为74%（95%CI：71%～76%）和99%（95%CI：98%～99%）;对儿童肺结核的诊断敏感度比成人肺结核低,74%（95%CI：65%～83%） vs 90%（95%CI：89%～92%）.Xpert MTB/RIF试验诊断耐多药结核病的敏感度为96%（95%CI：94%～97%）,特异度为98%（95%CI：98%～99%）.结论 Xpert MTB/RIF试验诊断结核病的价值较高,尤其是成人结核病及耐多药结核病.Xpert MTB/RIF试验在儿童结核病中的诊断价值由于纳入文献较少,尚待进一步研究.     

Xpert MTB/RIF、AFB与IGRA对结核病诊断价值的比较

目的 分析结核分枝杆菌rpoB基因和突变检测(Xpert MTB/RIF)、涂片抗酸染色镜检(ACID-fast bacilli smear microscopy, AFB)与γ干扰素释放实验(interferon-gamma release assay, IGRA)3种实验方法诊断结核病的价值及差异。方法收集北京同仁医院2019年1月至2021年3月同时检测Xpert MTB/RIF、AFB以及IGRA的患者1 400例,以中国国家行业标准WS288-2017作为结核病诊断标准,评价3种检测方法及差异。结果 1 400例患者中确诊结核病132例,其中肺结核116例,肺外结核16例。Xpert MTB/RIF阳性107例,灵敏度72.7%,特异性为100%;AFB阳性55例,灵敏度为33.3%,特异性为99.1%;IGRA阳性316例,灵敏度为89.4%,特异性为45.3%。结论 Xpert MTB/RIF特异性最高,灵敏度较高;IGRA灵敏度最高,特异性较低;AFB灵敏度较低,特异性较高,3种方法各有特点,联合使用有助于临床结核病的诊断。     

荧光PCR法检测石蜡标本中结核分枝杆菌的流程改进

<正>结核分枝杆菌的检测对于有肉芽肿性炎特征组织病变的病理诊断具有重要的参考意义。目前,病理科常用的检测方法有基于细菌学的抗酸染色、金胺O荧光染色,基于分子病理学的PCR、Xpert MTB/RIF检测^[1-2]等。其中荧光PCR技术具有敏感性高、特异性好、速度快,且价格实惠的特点,被广泛应用于大中型医院病理科福尔马林固定石蜡包埋（formalin-fixed paraffin-embedded,FFPE）组织的结核分枝杆菌的检测中^[3-6]。由于石蜡组织标本中的病菌分布不稳定,     

三种方法在盆腔结核诊断中的价值分析

目的 评价盆腔积液改良抗酸染色法、离心涂片法和外周血结核分枝杆菌感染T细胞斑点试验(T-SPOT.TB)在盆腔结核诊断中的价值.方法 回顾性分析确诊的35例盆腔结核患者和35例盆腔恶性肿瘤患者盆腔积液(对照组)的改良抗酸染色法和离心涂片法以及血液T-SPOT.TB检测结果.比较三种方法的灵敏度及特异度.结果 改良抗酸染色法敏感性为31.43％ (11/35),特异性100％;离心涂片法敏感性5.71％ (2/35),特异性100％;T-SPOT.TB敏感性为48.57％(17/35),特异性94.29％,改良抗酸染色法和T-SPOT.TB两种方法联合检测的阳性率为57.14％ (20/35).改良抗酸染色法与离心涂片法及T-SPOT.TB比较,其阳性率较高,差异均有统计学意义(P＜0.05),改良法与T-SPOT.TB联合检测阳性率较改良法高(P＜0.05).结论 改良抗酸染色法和T-SPOT.TB检测阳性率高,联合检测诊断效果更好,两种技术对盆腔结核有重要的辅助诊断价值.     

改良抗酸染色法诊断盆腔结核意义分析

目的 建立改良抗酸染色法并分析其在盆腔结核中的诊断价值.方法 对48例盆腔结核患者盆腔积液用离心涂片法、改良抗酸染色法和BACTEC MGIT 960培养法进行检测,并对结果进行分析.结果 离心涂片法、改良抗酸染色法和BACTEC MGIT 960培养法阳性率分别为2.08％ (1/48)、39.58％ (19/48)和37.50％ (18/48),改良抗酸法分别与离心涂片法及BACTEC MGIT 960培养法比较,改良抗酸染色法的阳性检出率最高,其与离心涂片法差异有统计学意义(P＜0.001),但与BACTEC MGIT 960培养法差异无统计学意义(P＞0.05).结论 改良抗酸染色法较离心涂片法阳性率明显提高,虽然和BACTEC MGIT 960培养法阳性率无明显差异,但是改良抗酸染色法可以快速诊断盆腔结核,对于临床工作有重要意义.     

恒温扩增检测结核分枝杆菌特异性IS6110片段在肺结核检测中的应用

目的 比较恒温扩增技术检测、抗酸杆菌涂片、结核分枝杆菌培养在肺结核患者检测中的应用.方法 收集2016年1月1日至12月31日我院呼吸科肺结核患者清晨痰液,将标本分为3份,分别进行痰涂片抗酸染色,结核菌培养及恒温扩增检测结核分枝杆菌特异性IS6110片段,统计分析3种方法的检测结果.结果 56例痰标本,恒温扩增检测结核分枝杆菌特异性IS6110片段阳性33例,阳性率为58.93％,抗酸染色阳性15例,阳性率为26.79％,恒温扩增检测结核分枝杆菌特异性IS6110片段阳性率高于抗酸染色方法;结核分枝杆菌培养阳性为35例,阳性率62.50％;恒温扩增检测结核分枝杆菌特异性IS6110片段和结核分枝杆菌培养两种方法检测Kappa系数为0.776,检测一致性较好.结论 恒温扩增检测结核分枝杆菌特异性IS6110片段是一种简便、高灵敏度的检出结核分枝杆菌的方法.     

Diagnostic Accuracy of Xpert MTB/RIF Assay in Extrapulmonary Tuberculosis

Introduction: The WHO endorsed Xpert Mycobacterium tuberculosis/rifampicin (MTB/RIF) assay, has been evaluated for pulmonary TB in a number of studies but very few have investigated it for extrapulmonary specimens. The present study evaluates the performance of Xpert MTB/RIF assay in the diagnosis of extrapulmonary TB (EPTB). Aim and Objectives: The aim of the study is to determine sensitivity and specificity of Xpert MTB/RIF assay for diagnosis of EPTB and RIF resistance in comparison to culture on Lowenstein–Jensen (LJ) medium and proportion method (PM), respectively. Materials and Methods: A total of 738 specimens from clinically suspected cases of EPTB were subjected to Ziehl–Neelsen staining, Xpert MTB/RIF assay and culture on LJ medium. PM was done on MTB isolates. Results: The sensitivity, specificity of Xpert MTB/RIF assay for diagnosis of EPTB were 84.91% (95% confidence interval [CI] 72.41%–93.25%) and 86.72% (95% CI 83.94%–89.17%) and for RIF resistance detection were 60.00% (95% CI 32.29%–83.66%) and 94.74% (95% CI 73.97%–99.87%), respectively. Among culture-positive cases, the sensitivity of Xpert MTB/RIF assay was 94.12% in smear positive and 80.56% in smear-negative cases. Xpert MTB/RIF showed maximum sensitivity of MTB detection from lymph node specimens (100% [95% CI 54.07%–100.00%]) and other body fluids (100% [95% CI 15.81%–100.00%]). Conclusion: The present study establishes Xpert MTB/RIF assay as a promising tool in the rapid diagnosis of EPTB and detection of RIF resistance.     

Xpert MTB/RIF assay for diagnosis of pleural tuberculosis

We prospectively investigated the diagnostic utility of the Xpert MTB/RIF (Mycobacterium tuberculosis/rifampin [RIF] resistance) assay in 20 cases with confirmed tuberculous pleural effusion. The sensitivity and specificity of the Xpert assay in pleural fluid were 25% and 100%, respectively. All cases positive by the Xpert assay were also positive by pleural fluid culture.     

Comparison of the Xpert MTB/RIF test with an IS6110-TaqMan real-time PCR assay for direct detection of Mycobacterium tuberculosis in respiratory and nonrespiratory specimens

The sensitivities of the Xpert MTB/RIF test and an in-house IS6110-based real-time PCR using TaqMan probes (IS6110-TaqMan assay) for the detection of Mycobacterium tuberculosis complex (MTBC) DNA were compared by use of 117 clinical specimens (97 culture positive and 20 culture negative for MTBC) that were frozen in sediment. The 97 clinical specimens included 60 respiratory and 37 nonrespiratory specimens distributed into 36 smear-positive and 61 smear-negative specimens. Among the 97 culture-positive specimens, 4 had rifampin-resistant isolates. Both methods were highly specific and exhibited excellent sensitivity (100%) with smear-positive specimens. The sensitivity of the Xpert MTB/RIF test with the whole smear-negative specimens was more reduced than that of the IS6110-TaqMan assay (48 versus 69%, P = 0.005). Both methods exhibited similar sensitivities with smear-negative respiratory specimens, but the Xpert MTB/RIF test had lower sensitivity with smear-negative nonrespiratory specimens than the IS6110-TaqMan assay (37 versus 71%, P = 0.013). Finally, the sensitivities of the Xpert MTB/RIF test and the IS6110-TaqMan assay were 79% and 84%, respectively, with respiratory specimens and 53% and 78%, respectively (P = 0.013), with nonrespiratory specimens. The Xpert MTB/RIF test correctly detected the rifampin resistance in smear-positive specimens but not in the one smear-negative specimen. The Xpert MTB/RIF test is a simple rapid method well adapted to a routine laboratory that appeared to be as sensitive as the IS6110-TaqMan assay with respiratory specimens but less sensitive with paucibacillary specimens, such as smear-negative nonrespiratory specimens.     

Improved Case Detection Using Xpert Mycobacterium tuberculosis/Rifampicin Assay in Skeletal Tuberculosis

Background: In India, musculoskeletal tuberculosis (TB) accounts for 10%–25% of extrapulmonary TB. Data on drug-resistant skeletal TB are lacking. At present, the diagnosis is based mainly on radiological techniques. Laboratory confirmation of skeletal TB is delayed as 6–8 weeks are required for culture results. Xpert Mycobacterium tuberculosis/rifampicin (MTB/RIF) assay is a fully automated test which simultaneously detects MTB and RIF resistance within 3 h. Hence, this study was done to compare the yield of case detection using Xpert assay in comparison with culture in specimens received from clinically suspected skeletal TB cases. Methods: Retrospective analysis of microscopy, culture and Xpert assay results was carried out on specimens received in laboratory from skeletal TB cases from January 2016 to December 2017. Results: Of the 201 patients analysed, majority of the specimens were obtained from the spine (55.72%). MTB was detected in 48.68% of tissue and 24% of pus specimens. Xpert assay was detected MTB in 67 (33.33%) specimens of which 53 (47.32%) were from the spine. Culture was detected MTB in 66 (32.83%) specimens. Xpert assay was detected two specimens more than culture. One specimen was positive by only culture. RIF-resistant MTB was detected in 10 (14.92%) specimens by Xpert assay. Conclusion: The spine is the most common site involved. Tissue specimen is better for early diagnosis. High RIF resistance in skeletal TB is an alarming situation. Ability of Xpert MTB/RIF assay for rapid and simultaneous detection of MTB and RIF resistance in comparison with culture makes it a useful diagnostic tool in skeletal TB.     

Comparison of two nucleic acid amplification assays, the Xpert MTB/RIF assay and the amplified Mycobacterium Tuberculosis Direct assay, for detection of Mycobacterium tuberculosis in respiratory and nonrespiratory specimens

We compared the performance of the Xpert MTB/RIF assay, a new real-time tuberculosis (TB) PCR test, with that of the Amplified Mycobacterium Tuberculosis Direct (MTD) assay using 162 respiratory and nonrespiratory specimens. Based on culture as the gold standard, the overall sensitivity and specificity for all sample types for the Xpert MTB/RIF assay were 90.9 and 89%, respectively, while for the MTD assay, the overall sensitivity and specificity were 97.3 and 87.1%, respectively. A higher proportion of total equivocal results were obtained for the MTD assay, at 10.5% (17/162), while the Xpert MTB/RIF assay generated 5.5% (9/162) of invalid reads.     

Evaluation of the Xpert MTB/RIF test accuracy for diagnosis of tuberculosis in areas with a moderate tuberculosis burden

The Xpert MTB/RIF assay (Xpert) is a molecular assay used for direct detection of Mycobacterium tuberculosis (MTB) in clinical specimens. In this study, we aimed to assess the accuracy of the Xpert assay for the diagnosis of tuberculosis (TB) in TB suspected patients from the northern region of Iran. The obtained results were compared with the culture method. The sputum specimens were examined using the Xpert assay, smear microscopy, and solid culture media as a reference diagnostic tool. Among 293 presumptive TB cases, 92 (31.4%) were positive according to the culture method. The Xpert method detected 88 (95.7%) cases that were positive according to the culture method, compared with 78 (84.8%) positive cases according to smear microscopy. The overall sensitivity and specificity of the Xpert method for TB diagnosis were 95.7% and 99%, respectively. Also, the sensitivity and specificity for smear microscopy were 84.8% and 97.5%, respectively. The Xpert assay showed high overall sensitivity and specificity; thus, it can be effectively used for the early and accurate diagnosis of MTB in TB endemic areas. In addition, the agreement between semi‐quantitative results of Xpert and smear microscopy assays could be helpful in evaluating transmission potential in TB patients.     

GeneXpertMTB/RIF Assay对肺部结核的临床诊断及治疗价值

目的:探究GeneXpertMTB/RIF Assay对临床诊断及治疗肺部结核的应用价值.方法:收集2019年1月至2019年12月在甘孜藏族自治州人民医院住院的肺结核患者的痰液标本148份,同时选取50份非结核肺部感染患者的痰液标本作为对照.分别采用涂片抗酸杆菌检查、MGIT960检测MTB及药敏法和GeneXper...     

Dried culture spots for Xpert MTB/RIF external quality assessment: results of a phase 1 pilot study in South Africa

Implementation of Xpert MTB/RIF requires quality assessment. A pilot program using dried culture spots (DCSs) of inactivated Mycobacterium tuberculosis is described. Of 274 DCS results received, 2.19% generated errors; the remainder yielded 100% correct Mycobacterium tuberculosis detection. The probe A cycle threshold (C(T)) variability of three DCS batches was ≤ 3.47. The study of longer-term DCS stability is ongoing.     

Comparison of the Xpert MTB/RIF and Cobas TaqMan MTB Assays for Detection of Mycobacterium tuberculosis in Respiratory Specimens

The Xpert MTB/RIF assay (Cepheid, Sunnyvale, CA) is a fully automated, cartridge-based real-time PCR assay designed to detect Mycobacterium tuberculosis and rifampin resistance within 2 h. The performance of the Xpert assay has been evaluated in various clinical settings. However, there are few data comparing the Xpert assay to the Cobas TaqMan MTB test (Roche Diagnostics, Basel, Switzerland), one of the most widely utilized molecular assays for M. tuberculosis detection. In this prospective study, 320 consecutive respiratory specimens were processed simultaneously using acid-fast bacillus (AFB) staining, mycobacterial cultures with both solid and liquid media, and the Cobas and Xpert assays. The Xpert assay was performed with direct respiratory specimens, while the Cobas assay was done with decontaminated concentrated specimens. Based on the culture as a reference method, the overall sensitivities of the Cobas and Xpert assays were 71.4% and 67.9%, respectively. When AFB smear results were taken into consideration, the sensitivities of the Cobas assay for smear-positive and -negative specimens were 87% and 54%, while those of the Xpert assay were 67% and 69%, respectively. The Cobas assay showed 100% specificity and 100% positive predictive value (PPV) regardless of smear results, while the Xpert assay showed 100% specificity and 100% PPV for smear-positive specimens but 98% specificity and 60% PPV for smear-negative specimens. In conclusion, the Xpert assay showed performance that was slightly inferior to that of the Cobas assay but seems useful for the rapid detection of M. tuberculosis, considering that it was performed without laborious and time-consuming decontamination and concentration procedures.