Xpert MTB/RIF在骨关节结核患者快速诊断中的应用

目的:探讨Xpert MTB/RIF在骨关节结核患者快速诊断中的应用价值。方法:2014年2月~2014年11月使用Xpert MTB/RIF对49例骨关节结核患者及32例非结核性骨关节病患者的脓液标本进行检测,以临床诊断为金标准,计算Xpert MTB/RIF检测结核分枝杆菌的敏感性、特异性、阳性预测值、阴性预测值、一致率;同时对脓液标本行抗酸染色及结核分枝杆菌快速培养(BACTECT MGIT960),比较Xpert MTB/RIF与抗酸染色、结核分枝杆菌快速培养在敏感性及特异性上的差异;综合上述两方面因素评价Xpert MTB/RIF在骨关节结核患者快速诊断中的应用价值。结果:Xpert MTB/RIF检测单个脓液标本的时间为2.3±0.2h。49例骨关节结核患者脓液标本中,46例Xpert MTB/RIF检测结核分枝杆菌阳性,3例Xpert MTB/RIF阴性;32例非结核性骨关节病患者脓液标本中,1例Xpert MTB/RIF检测结核分枝杆菌阳性,31例Xpert MTB/RIF阴性;以临床诊断为金标准,Xpert MTB/RIF检测结核分枝杆菌的敏感性为93.87%、特异性为96.87%、阳性预测值为97.87%、阴性预测值为91.17%,一致率为95.06%。在46例Xpert MTB/RIF结核分枝杆菌阳性的骨关节结核患者中,10例存在利福平耐药突变基因,耐药突变率为21.73%。49例骨关节结核患者脓液标本中,抗酸染色阳性8例,阴性41例,敏感性为17.39%;结核分枝杆菌快速培养阳性患者11例,阴性38例,敏感性为23.91%;32例非结核性骨关节病患者脓液标本抗酸染色及结核分枝杆菌快速培养均为阴性,其特异性均为100%。Xpert MTB/RIF检测结核分枝杆菌的的敏感性优于抗酸染色及结核分枝杆菌快速培养(P0.05),特异性与抗酸染色及结核分枝杆菌快速培养无明显差异(P0.05)。结论:Xpert MTB/RIF在骨关节结核患者的快速诊断中具有较高的诊断效能,其耗时短,敏感性高,特异性强,与抗酸染色及结核分枝杆菌快速培养比较具有明显优势。     

Xpert MTB/RIF在骨关节结核患者

【摘要】 目的：探讨Xpert MTB/RIF在骨关节结核患者快速诊断中的应用价值。方法：2014年2月~2014年11月使用Xpert MTB/RIF对49例骨关节结核患者及32例非结核性骨关节病患者的脓液标本进行检测,以临床诊断为金标准,计算Xpert MTB/RIF检测结核分枝杆菌的敏感性、特异性、阳性预测值、阴性预测值、一致率;同时对脓液标本行抗酸染色及结核分枝杆菌快速培养（BACTECT MGIT 960）,比较Xpert MTB/RIF与抗酸染色、结核分枝杆菌快速培养在敏感性及特异性上的差异;综合上述两方面因素评价Xpert MTB/RIF在骨关节结核患者快速诊断中的应用价值。结果：Xpert MTB/RIF检测单个脓液标本的时间为2.3±0.2h。49例骨关节结核患者脓液标本中,46例Xpert MTB/RIF检测结核分枝杆菌阳性,3例Xpert MTB/RIF阴性;32例非结核性骨关节病患者脓液标本中,1例Xpert MTB/RIF检测结核分枝杆菌阳性,31例Xpert MTB/RIF阴性;以临床诊断为金标准,Xpert MTB/RIF检测结核分枝杆菌的敏感性为93.87%、特异性为96.87%、阳性预测值为97.87%、阴性预测值为91.17%,一致率为95.06%。在46例Xpert MTB/RIF结核分枝杆菌阳性的骨关节结核患者中,10例存在利福平耐药突变基因,耐药突变率为21.73%。49例骨关节结核患者脓液标本中,抗酸染色阳性8例,阴性41例,敏感性为17.39%;结核分枝杆菌快速培养阳性患者11例,阴性38例,敏感性为23.91%;32例非结核性骨关节病患者脓液标本抗酸染色及结核分枝杆菌快速培养均为阴性,其特异性均为100%。Xpert MTB/RIF检测结核分枝杆菌的的敏感性优于抗酸染色及结核分枝杆菌快速培养（P<0.05）,特异性与抗酸染色及结核分枝杆菌快速培养无明显差异（P>0.05）。结论：Xpert MTB/RIF在骨关节结核患者的快速诊断中具有较高的诊断效能,其耗时短,敏感性高,特异性强,与抗酸染色及结核分枝杆菌快速培养比较具有明显优势。     

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。     

结核分枝杆菌/利福平耐药实时荧光定量核酸扩增检测技术在脊柱结核检测中的应用

目的 探讨结核分枝杆菌/利福平耐药实时荧光定量核酸扩增检测技术(GeneXpert Mycobacterium tuberculosis/Ri-fampin,Xpert MTB/RIF)在不同类型脊柱结核组织标本检测中的作用.方法 收集127例脊柱结核患者的301份标本(脓液108份、干酪样标本94份、肉芽组织99份)进行细菌培养和Xpert MTB/RIF检测,计算阳性率.结果 127例中,培养阳性60例,阳性率47.24％;301份标本中培养阳性138份,阳性率45.85％.脓液标本和干酪样标本培养阳性率均高于肉芽组织(P<0.05).Xpert MTB/RIF检测脓液、干酪样标本和总标本阳性率高于细菌培养(P<0.05);60例细菌培养阳性的患者中,Xpert MTB/RIF法的敏感度为96.67％;利福平耐药27例,耐药率为45％,Xpert MTB/RIF检测利福平的敏感度为96.30％,特异性为94.29％;67例细菌培养阴性的患者中,36例Xpert MTB/RIF法检测阳性,敏感度为53.73％.结论 Xpert MTB/RIF在脊柱结核标本检验中有较高的敏感性,收集脊柱结核患者的各种标本尤其是脓液标本,使用Xpert MTB/RIF可方便、快速、准确地诊断结核感染和耐药,值得临床推广应用.     

GeneXpert法检测结核分枝杆菌及其对利福平耐药性的研究

目的探讨GeneXpert MTB/RIF(简称Xpert)法检测结核分枝杆菌及其利福平耐药性的可行性。方法选取2013年3月至6月在本院就诊的413例疑似肺结核病患者,对其痰标本分别进行抗酸染色镜检、培养、比例法体外药敏试验和Xpert法检测,并以传统的罗氏固体培养和比例法药敏试验为金标准,对Xpert法检测结果进行分析。结果以培养法为金标准,Xpert法检测197例涂阴患者的敏感性为55.2%,特异性为93.5%,与培养法检测结果差异无统计学意义(χ2=0.04,P=0.841);Xpert检测214例涂阳患者的敏感性97.5%,特异性54.5%,与培养法检测结果差异也无统计学意义(χ2=0.10,P=0.752)。以比例法药敏为金标准,对214例培养阳性的患者,Xpert法检测利福平耐药的敏感性为95.3%,特异性为99.4%,与比例法检测结果差异无统计学意义(χ2=0.00,P=1.000)。结论Xpert法适用于结核分枝杆菌及耐药结核分枝杆菌的快速筛查。     

结核分枝杆菌相关γ-干扰素定量检测在脊柱结核诊断中的应用

目的探讨结核分枝杆菌相关γ-干扰素定量检测(TB-IGRA)在脊柱结核诊断中的应用价值。方法分析44例疑似脊柱结核病例资料,结合最终临床诊断,分为脊柱结核组和非脊柱结核组,计算TB-IGRA、抗结核抗体(TB-Ab)检测、利福平耐药实时荧光定量核酸扩增检测(Xpert MTB/RIF)及病理学检测4种方法诊断脊柱结核的灵敏度、特异度、阳性预测值和阴性预测值,采用kappa一致性检验评估TB-IGRA与TB-Ab检测、Xpert MTB/RIF及病理学检测结果的一致性。结果纳入的44例患者中,确诊或高度怀疑脊柱结核病28例,确诊非脊柱结核16例。TB-IGRA的灵敏度、特异度、阳性预测值和阴性预测值分别为85.7%、68.8%、82.8%和73.3%;TB-Ab检测的灵敏度、特异度、阳性预测值和阴性预测值分别为53.6%、87.5%、88.2%和51.9%;Xpert MTB/RIF的灵敏度、特异度、阳性预测值和阴性预测值分别为68.8%、71.4%、84.6%和50.0%;病理学检测的灵敏度、特异度、阳性预测值和阴性预测值分别为92.9%、100%、100%和88.9%。TB-IGRA与TB-Ab诊断结果一致性较差(κ=0.25),与Xpert MTB/RIF及病理学诊断结果一致性较高(κ=0.68、0.71)。结论TB-IGRA作为脊柱结核的辅助诊断手段,简便、快速,灵敏度和特异度较高,尤其适用于不方便获取病灶组织的患者。     

荧光定量PCR技术检测肺外结核患者样本临床应用

目的评估赛沛分子检测技术（结核分枝杆菌和利福平耐药基因）（GeneXpert MTB/RIF）检测心包和胸腔积液样本中结核分枝杆菌的有效性。 方法收集2015年1月至2017年6月天门市第一人民人民医院和汉川市人民医院收治的286例结核病疑似患者的临床资料,分别收集158例胸腔积液和128例心包积液样品。每个样品均经过抗酸染色涂片镜检、罗氏培养基结核分枝杆菌培养（LJ培养）和GeneXpert MTB/RIF测定。使用结核分枝杆菌罗氏培养作为金标准,评估GeneXpert MTB/RIF技术检测MTB的有效性。 结果在286例积液样本中,MTB通过LJ培养阳性者51例（17.8%）,GeneXpert MTB/RIF测定阳性者43例（15%）,抗酸染色镜检阳性者11例（3.8%）。GeneXpert技术灵敏度、特异性、阳性预测值和阴性预测值分别为84.3%、100%、100%和96.7%,抗酸染色方法的灵敏度、特异性、阳性预测值和阴性预测值分别为18.3%、99.1%、81.8%和85.4%。GeneXpert技术检查心包积液中MTB的灵敏度可达90%。GeneXpert MTB/RIF和抗酸染色镜检鉴定两种样本中结核分枝杆菌有效性的差异具有统计学意义（χ² = 233.199、33.715、P均< 0.001）。 结论GeneXpert MTB/RIF技术对于检测胸腔和心包积液中的MTB具有高灵敏度和高特异性。     

核酸检测指导个体化耐药脊柱结核治疗

[目的]评价Xpert MTB/RIF指导的个体化化疗治疗耐利福平/耐多药脊柱结核的初步临床疗效.[方法]回顾分析2017年6月～2018年6月120例接受手术治疗的脊柱结核患者,术后参照既往抗结核化疗史及Xpert MTB/RIF检测结果,制定个体化化疗方案,待表型药敏结果回示后根据药敏结果再次调整化疗方案.[结果]...     

结核分枝杆菌快速培养和常规药敏试验在脊柱结核治疗中的应用

目的 探讨BACT/ALERT 3D系统快速培养和绝对浓度法药敏试验对指导脊柱结核个体化化疗的应用价值,分析研究脊柱结核耐药情况.方法 根据临床表现、影像学表现、病理检查,50例患者诊断为脊柱结核,并接受手术治疗.收集术中所取脓液、干酪样组织.低温避光保存,8 h内送检,常规处理后接种液体培养基,使用BACT/ALERT 3D系统进行分枝杆菌快速培养.培养阳性者接种PNB和TCH培养基进行菌种鉴定,并将细菌接种至含药改良罗氏培养基,按绝对浓度法进行11种常用一线和二线抗结核药物药敏试验.结果 50例标本培养阳性21例(42%),人型结核杆菌19例,牛型结核杆菌2例.结核杆菌培养和药敏试验平均耗时41 d(28～58 d).其中耐药11例(52.4%),异烟肼耐药4例(19.0%),利福平和乙胺丁醇各1例(4.8%),链霉素3例(14.3%),力克肺疾2例(9.5%),左氧氟沙星8例(38.1%).结论 结核分枝杆菌快速培养和常规药敏试验准确度高,费用低,可检测常用一线和二线药物的敏感性,适用于指导脊柱结核个体化化疗方案的制定.异烟肼、利福平、吡嗪酰胺、乙胺丁醇或(和)链霉素联合用药方案对多数初治脊柱结核患者有效.     

二代测序技术应用于脑脊液检测在结核性脑膜炎中的早期诊断价值

目的评价二代测序技术应用于脑脊液检测在结核性脑膜炎（TBM）患者中的早期诊断价值。 方法前瞻性纳入2018年2月2日至2018年8月2日于山东省胸科医院就诊的临床怀疑TBM的患者共50例,并跟踪随访其诊疗结局。送检脑脊液标本均进行二代测序,测序所得原始序列与病原微生物数据库进行对比得到最终结果。二代测序结果以检测到结核分枝杆菌复合群唯一比对序列为阳性,未检测到唯一比对序列为阴性。以符合脑脊液结核分枝杆菌培养阳性、涂片阳性、Xpert MTB/RIF检测阳性及结核分枝杆菌核酸检测阳性等4项中至少1项即为确诊TBM患者;临床可疑TBM且抗结核治疗有效为临床诊断患者;有其他病原学依据或临床排除TBM者为非TBM患者。分析二代测序在TBM早期诊断中的敏感性和特异度。 结果确诊为TBM患者22例中Xpert MTB/RIF检测阳性13例,培养阳性6例,结核分枝杆菌核酸PCR检测阳性5例,临床诊断为TBM患者12例,非TBM患者16例。在确诊及临床诊断患者中,二代测序技术检测到结核分枝杆菌复合群系列20例,敏感性为58.8%（20/34）,特异度为100%（16/16）。在确诊患者中,二代测序的敏感性为63.6%（14/22）;在同步进行结核分枝杆菌培养、Xpert MTB/RIF检测与二代测序的50例标本中,以临床诊断为标准,3种方法的特异度均为100%（16/16）;传统方法、Xpert MTB/RIF检测及二代测序的敏感性分别为29.4%（10/34）、38.2（13/24）和58.8（20/34）,前两种检测方法与二代测序敏感性差异均有统计学意义（McNemar检验：χ² = 8.333、P = 0.013,χ² = 8.333、P = 0.065）。传统方法与二代测序联合检测的敏感性高达82.4%（28/34）。 结论二代测序技术能够较快速地检测脑脊液中的结核分枝杆菌复合群,且其敏感性和特异度均较高,可作为TBM的早期诊断指标。二代测序联合传统检测方法可提高检出率。     

Gene Xpert/MTB RIF assay for spinal tuberculosis- sensitivity,specificity and clinical utility

BackgroundXpert MTB/RIF assay is a rapid automated molecular test with excellent reported sensitivity, specificity for diagnosis of pulmonary and extrapulmonary Mycobacterium tuberculosis (MTB) infections. However, the clinical utility and accuracy in STB is not well established. A study was conducted to report on the sensitivity, specificity and clinical utility of the Xpert MTB/RIF assay in spinal tuberculosis (STB).MethodsA retrospective review of medical records was performed for 136 patients that underwent spinal biopsy for suspected spondylodiscitis. Reports for acid fast bacilli (AFB) smear, gram stain, pyogenic culture, MTB culture, histopathology, Xpert MTB/RIF assay, and drug sensitivity testing were reviewed. ‘Reference standard for diagnosis of STB’ was based on positive histopathology and/or MTB culture evidence and was considered as MTB positive. Any samples returning a positive pyogenic or fungal culture were considered as MTB negative. The sensitivity, specificity for Xpert MTB/RIF was assessed against the reference standard.ResultsA total of 125 patients were considered for final analysis, 86 patients met the criteria for ‘Reference standard for diagnosis of TB spine’ (MTB positive). This includes nine patients that were MGIT culture only positive; 45 that were histopathology only positive and 32 were both culture and histopathology positive. There were 39 culture proven (pyogenic-37 and fungal-2) patients included in MTB negative group. The 86 MTB positive patients, included 53 (61.6%) tissue samples and 33 (38.4%) pus samples. The overall analysis showed a 65.1% sensitivity, 100% specificity, 100% PPV and 56.5% NPV for the Xpert MTB/RIF.ConclusionsGene Xpert MTB/RIF showed excellent specificity and was accurate in the identification of drug resistance. The sensitivity was 65% and sampling techniques using pus samples rather than tissue samples could be a possible reason for lower sensitivity.     

The role of Xpert MTB/RIF assay in the diagnosis of tubercular spondylodiscitis

Purpose

This study aims to assess the accuracy of the Xpert MTB/RIF assay in the diagnosis of tubercular spondylodiscitis and to identify its role in detecting Rifampicin resistance in patients with infective spondylodiscitis.

Methods

A retrospective study including 348 patients suspected to have infective spondylodiscitis was done. Tissue/pus samples obtained were sent for culture, histopathology and Xpert MTB/RIF assay. All patients who were confirmed to have tubercular spondylodiscitis and those patients who were suspected on clinico-radiological basis were also treated with anti-tuberculous chemotherapy for a period of 9 months. The efficacy of the Xpert MTB/RIF assay was assessed in terms of sensitivity and specificity when compared to culture, histopathology, and Composite reference standard (CRS).

Results

During this study period of 24 months, a total of 348 patients were treated for infective spondylodiscitis. 254 patients were treated for tuberculosis following a smear positivity, culture positivity, and histopathology report or empirically based on clinico-radiological findings. The sensitivity and specificity of the Xpert MTB/RIF assay when compared to culture were 88.4 and 63.7%, respectively. When compared to both culture and histopathology reports it was 80.9 and 80.6%. The sensitivity and specificity of the Xpert MTB/RIF assay when compared to composite reference standard were 71.2 and 100%, respectively. The sensitivity of the assay to detect Rifampicin resistance was 100%. The prevalence of Rifampicin resistance was 5.1%.

Conclusion

This study recommends Xpert MTB/RIF assay for early detection of Mycobacterium tubercular spondylodiscitis and Rifampicin resistance.

     

Spectrum of Drug Resistance in Musculoskeletal Tuberculosis

BackgroundVery few studies report resistance pattern exclusively in musculoskeletal tuberculosis (MSK-TB).MethodsThis study of 100 pus samples from patients of MSK-TB with active disease in whom Mycobacterium tuberculosis (MTB) was detected by cartridge-based nucleic acid amplification test (CBNAAT), revealed the pattern of resistance among newly diagnosed and previously treated cases. Liquid culture and drug susceptibility testing (DST) using MGIT 960 was done for 11 anti-tubercular drugs.ResultsAmong these 100 cases; 22% were AFB positive; MGIT 960 detected MTB in 58.33% (35/60) new cases and 30.0% (12/40) previously treated cases. Five new and 10 previously treated cases had drug resistance and 12 were detected rifampicin resistance (Rif-R) by CBNAAT. Among new cases MGIT-DST detected mono-INH resistant in 2.86% (1/35), mono-STR resistant in 2.86% (1/35), MDR-TB in 5.7% (2/35) and pre-XDR in 2.9%(1/35).Among previously treated cases Rif-R was found in 10% (4/40) where MTB was not detected by MGIT and MGIT-DST detected mono-INH resistant in 8.33% (1/12); MDR-TB in 8.33% (1/12) and pre-XDR in 33.3%. There were no cases of XDR-TB.ConclusionHigh disease burden of various type drug resistance were seen more commonly in previously treated cases and was not uncommon in new cases of MSK-TB. Both CBNAAT and DST are essential for detecting resistance pattern in MSK-TB.     

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。     

基于85B mRNA的逆转录-实时定量聚合酶链反应对肺结核分枝杆菌感染的诊断价值

目的探讨痰液样本中检测结核分枝杆菌（MTB）85B mRNA的逆转录-实时定量聚合酶链反应（RT-qPCR）对MTB感染的诊断价值。 方法分别纳入2016年1月至2016年12月陕西省结核病防治院经痰培养法（BACTEC MGIT 960系统）确诊为MTB阳性患者60例为试验组,60例无MTB感染的健康人为对照组。提取痰液样本核酸,分别采用TaqMan法检测MTB基因中编码16S rRNA的DNA序列和RT-qPCR法检测MTB的85B mRNA,比较两种方法的诊断效能（敏感性、特异性、阳性预测值、阴性预测值和准确性）。 结果TaqMan法诊断MTB感染的敏感性、特异性、阳性预测值、阴性预测值和准确性分别为93.3%、83.3%、84.9%、92.6%和88.3%;RT-qPCR法分别为98.3%、95.0%、95.2%、98.3%和96.7%。通过二元Logistics回归分析证实基于85B mRNA的RT-qPCR法对MTB感染鉴别能力更强（OR = 19.924,95%CI：5.364～73.012,P < 0.001）。 结论基于结核分枝杆菌85B mRNA的RT-qPCR方法能够快速、有效诊断MTB感染。     

Ileocaecal and transverse colonic tuberculosis mimicking colonic malignancy – A case report

IntroductionGastrointestinal tuberculosis is common in the developing world especially in the lower socioeconomic groups. In elderly, it may mimic malignancy.Case presentationA 46-year-old female presented with a 6 month history of diffuse pain in abdomen with low grade fever and loss of weight and appetite. Clinically, differential of malignancy of the large bowel was considered. The computerized tomography(CT) scan of the abdomen revealed a diffuse concentric long segmental thickening of terminal ileum, ileo ceacal junction, ascending colon and narrowing of the transverse colonic end of the splenic flexure suggesting an infective etiology. Colonoscopy showed an ulcero-nodular lesion at the splenic flexure raising the possibility of colonic cancer and thickening of ascending colon and caecum. Colonoscopic biopsy from both sites, on histopathology, showed a moderate mixed inflammation and occasional lymphoid collection and crypt abscesses in the lamina propria giving a differential of tuberculosis or Crohn‘s disease. Biopsy smear showed occasional acid fast bacilli(AFBs) and the gene Xpert detected mycobacterium tuberculosis(MTB). The patient was started on anti Koch’s therapy(AKT).DiscussionIn this case the differential diagnosis was malignancy of the colon, inflammatory bowel disease and tuberculosis as all these conditions may have similar clinical profile and radiological findings. Tuberculosis of bowel was considered as the most probable diagnosis due to the CT findings. But the colonoscopy suggested malignant etiology.ConclusionPossibility of tuberculosis should be kept in mind while dealing with synchronous lesions in large intestine.