γ干扰素释放试验在肺内结核和肺外结核中的诊断价值

目的：探讨IFN-γ释放试验在肺内及肺外结核诊断中的应用价值。方法采用IFN-γ释放试验试剂盒分别检测185例结核病患者（包括119例肺结核和66例肺外结核）、139例其他肺部疾病患者和100例健康对照者结核患病情况,并以病原培养或/和临床诊断为标准,评价IFN-γ法诊断的敏感度和特异性,同时分别与痰菌培养及痰涂片平行检验的结果作比较。结果以临床诊断为准金标准,IFN-γ释放试验检测敏感度为93.51%,特异度为84.52%,检测肺结核和肺外结核的敏感度分别为90.76%和98.49%,两者差异无统计学意义（P〉0.05）;而痰涂片检测肺结核和肺外结核的敏感度分别为11.76%和3.03%;痰培养检测的敏感度也仅为24.37%和3.03%,3种方法对肺结核和肺外结核检测敏感度,差异均有统计学意义（ P均〈0.05）。全部424例标本,经培养阳性有33例,其中鉴定发现2例非结核分枝杆菌感染,均为脓肿分枝杆菌,IFN-γ释放试验检测为阴性,按鉴定确认的31例结核分支杆菌培养阳性结果为金标准,IFN-γ释放试验敏感度为90.32%（95%CI：75.10%~96.65%）。结论IFN-γ释放试验肺内、外结核具有快速方便和较高的敏感度,值得用于临床结核筛查。     

TaqMan-MGB荧光定量PCR快速检测痰结核分枝杆菌异烟肼耐药效果的评价

目的评价通过检测结核分枝杆菌katG基因突变以诊断异烟肼耐药并检测结核分枝杆菌的TaqMan-MGB荧光定量PCR方法。方法收集临床诊断为肺结核病的患者痰样本,用已建立的荧光PCR方法进行检测,与常规痰涂片抗酸染色、分枝杆菌培养和传统比例法药敏试验比较,评价检测结核分枝杆菌敏感性和特异性及异烟肼耐药的特异性和敏感性。统计学分析采用χ2检验,P值<0.05为差异有统计学意义。结果荧光定量PCR共检测186份痰液样本,敏感性为84.47%,特异性为100.00%。在确诊病人的痰液中阳性检出率84.47%,明显高于痰涂片40.99%(χ2=46.44,P<0.01),痰培养40.37%(χ2=47.89,P<0.01),且差异有统计学意义。其中荧光PCR在菌阳痰液中检出率94.44%,涂阴培阴的痰液中检出率75.28%。与常规药敏试验结果比较,培养阳性的61份痰样本的药敏符合率96.72%(59/61),敏感性33.33%(1/3),特异性100.00%(58/58)。结论 TaqMan-MGB荧光定量PCR的方法能特异、灵敏、快速诊断痰液中的结核分枝杆菌及其异烟肼耐药性。     

快速核酸检测系统用于肺结核临床诊疗的效果评价

【目的】通过比较确诊肺结核患者的实验室检测结果,评价快速核酸检测系统(GeneXpert MTB/RIF)在肺结核临床诊疗的应用效果。【方法】纳入2017年7月-2018年6月在上海市普陀区中心医院肺科门诊符合诊断标准的97名肺结核患者,开展痰涂片检查(Ziehl-Neelsen染色)、痰培养(MGIT液体培养)、XpertMTB/RIF检测,痰培养阳性者进行比例法药物敏感性试验和PNB法菌种鉴定,比较各方法的一致性。【结果】GeneXpert结核分枝杆菌检测与细菌学检测相比,灵敏度为93.44%,特异度为55.55%,检测一致率为79.38%。GeneXpert结核分枝杆菌检测与PNB法菌种鉴定的一致率为94.55%。GeneXpert耐利福平检测与比例法药敏比较,灵敏度为66.67%,特异度为98.08%,两方法的一致率为96.36%。【结论】GeneXpert MTB/RIF检测技术可与痰涂片和痰培养同时开展,提高确诊肺结核患者病原学阳性率,并尽早发现耐药肺结核患者。但仍需要进行菌种鉴定和药物敏感性试验,保证患者得到个体化治疗方案,获得最佳治疗效果。     

Problems in the standardization of the polymerase chain reaction for the diagnosis of pulmonary tuberculosis]

INTRODUCTION: The recent increase in the number of tuberculosis cases has called the world's attention once again to a perennial health problem, especially prevalent in developing countries. The time elapsed between the diagnosis and the institution of therapy is an obstacle to tuberculosis control and there is an urgent need for the development of techniques for the disease's rapid diagnosis. To achieve this goal, molecular biology techniques have been exhaustively investigated. This work describes the use of a polymerase chain reaction for rapid diagnosis of tuberculosis in a developing country. The sensitivity and specificity of this technique is compared to standard techniques used in the microbiology laboratory. METHODS: This study was undertaken in Ribeir?o Preto, S. Paulo State, Brazil. Forty-two sputum samples from suspected cases of tuberculosis attending the municipal health care centers were sent to the microbiology laboratory. The samples were processed for the detection of Mycobacterium tuberculosis by acid-fast bacilli determination, culture in Lowenstein-Jensen medium, and by a polymerase chain reaction that amplified a fragment of 123 base pairs of the Mycobacterium tuberculosis genome. RESULTS: Of the forty-two samples studied, one was contaminated and excluded from the study, ten were culture positive, ten were positive for the presence of acid-fast bacilli, and sixteen were polymerase chain reaction positive. The sensitivity and specificity of this technique were 90% and 81%, respectively. CONCLUSIONS: The polymerase chain reaction presented a sensitivity comparable to the culture and the whole procedure took only one day to complete. The results presented here make it a strong candidate for rapid diagnosis of tuberculosis in clinical settings making it possible to begin the specific therapy early in the course of the disease. However, standardization of the technique is necessary, and the correlation with clinical findings is of paramount importance due to the high sensitivity of this technique.     

环介导等温扩增法诊断初诊肺结核病研究

目的:初步评价环介导等温扩增法(Lamp)检测痰中结核分枝杆菌的准确性,探讨临床诊断初诊肺结核病人的应用价值。方法:选取249例初诊疑似肺结核病人的痰标本,同时进行Lamp、直接涂片抗酸染色法和改良罗氏培养基培养法检测。结果:剔除培养失败和非结核杆菌的标本,共检测了739份痰标本,其中直接涂片抗酸染色法检出阳性标本70份,阳性率9.47%;培养检出阳性标本118份,阳性率15.97%;Lamp检出阳性标本99份,阳性率13.40%。以培养法为标准,涂片法的灵敏度57.63%,特异度99.68%,符合率为94.30%;Lamp的灵敏度77.12%,特异度98.71%,符合率为96.61%。结论:Lamp在检测初诊疑似肺结核病人中有较高的灵敏度和特异性,适用于基层结核病工作中初诊肺结核病人的诊断。     

重庆市结核患者耐药结核杆菌embB306分子突变特征研究

目的 了解重庆结核杆菌的embB306突变特征,评价其作为诊断乙胺丁醇(ethambutol,EMB)耐药结核杆菌分子标记物的临床应用价值.方法 采用DNA测序方法分析重庆市结核患者痰中分离的51株EMB耐药结核杆菌和50株EMB敏感株的embB基因突变特征,与传统药敏实验进行诊断学试验评价embB306突变作为EMB耐药株分子标记物的临床应用价值.结果 embB306突变率在51株EMB耐药和50株敏感结核杆菌中分别为75.5%、6%;其中来自复治病例的EMB耐药菌株的embB306突变率是87.5%,高于来自初治病例的48.1%;embB306突变率随菌株耐药数量增加而升高,embB306在EMB耐药的耐多药菌株(Multidrug-Resistant Tuberculosis,MDR-TB)中的突变率高于EMB耐药的非MDR-TB和EMB敏感的MDR-TB的突变率;以embB306突变诊断EMB耐药菌株的诊断学试验评价主要指标为:灵敏度为66.7%;特异度为94.0%;准确度为80.2%;Youden指数为60.7%.结论embB306突变是重庆地区结核杆菌产生EMB耐药性的主要分子机制,且与耐药数量和治疗时间有关,其作为EMB耐药菌株诊断的分子标记物具有一定临床应用价值.     

口岸卫生检疫中开放性肺结核检测模式的探讨

目的探讨适合口岸开放性肺结核的检疫模式。方法对肺部透视、痰涂片(MTB)、痰聚合酶链式反应(PCR)、基因芯片(DNA-Chips)技术、结核菌素皮试(PPD)、抗脂阿拉伯甘露塘抗体 (LAM-IgG)实验、血清可溶性白细胞介素2受体(sIL-2R)实验、TB培养等检测方法的敏感性、特异性及检测周期等因素进行分析对比。结果以上各种检测方法在口岸开放性肺结核的检疫中各有优劣,但没有一种单独的检测方法可满足检疫目的和要求。结论本文建议为提高检疫有效性和明确检测的目的性应采用多种检测方法的不同的渐进式联合检测模式。     

评价显微镜观察药物敏感性技术(MODS)快速检测肺泡灌洗液中结核分枝杆菌对菌阴肺结核的临床诊断价值

目的:探讨肺泡灌洗液中结核分枝杆菌对菌阴肺结核临床诊断过程中显微镜观察药物敏感性技术(MODS)的应用价值。方法:整理2017年3月~2019年3月本院100份肺泡灌洗液标本,对其分别实施MODS、罗氏培养法检测,回顾分析检测结果。结果:与罗氏培养法相比,MODS对结核分枝杆菌的阳性检出时间明显缩短(P<0.05);MODS对结核分枝杆菌的阳性检出率高于罗氏培养法对应的阳性检出率(P<0.05)。结论:肺泡灌洗液中结核分枝杆菌对菌阴肺结核临床诊断期间通过应用MODS技术,可缩短诊断时间,提高阳性检出,保证菌阴性肺结核的诊断准确率。     

荧光定量PCR在结核分枝杆菌检测中的应用价值

目的比较荧光定量聚合酶链反应(FQ-PCR)与抗酸染色、改良罗氏培养法检测结核分枝杆菌的灵敏度和特异性,探讨FQ-PCR检测痰标本中结核分枝杆菌的应用价值。方法对311例临床确诊的肺结核病患者和40例非结核呼吸系统疾病患者的痰标本应用FQ-PCR法、痰涂片抗酸染色法、改良罗氏培养法检测结核分枝杆菌。结果FQ-PCR、抗酸染色法、改良罗氏培养法检测结核分枝杆菌的阳性率分别为47.0%(146/311)、28.3%(88/311)、35.4%(110/311),特异性分别为100.0%、100.0%、95.2%,以FQ-PCR检测的阳性率最高。结论FQ-PCR是一种快速、敏感性高、特异性强的结核分枝杆菌辅助诊断方法,在临床上有重要应用价值。     

RNA恒温扩增实时检测技术检测肺泡灌洗液对痰涂片阴性肺结核的快速诊断价值meta分析

目的 采用meta分析方法评价结核分枝杆菌RNA恒温扩增实时检测技术（simultaneous amplification and testing for Mycobacterium tuberculosis, SAT-TB）检测支气管肺泡灌洗液对痰涂片阴性肺结核的快速诊断价值。 方法 通过对中国知网,万方数据库,维普数据库,Pubmed,Embase,The Cochrane Library,Web of Science等数据库进行检索。检索符合要求的文献,对文献质量进行评价,提取文献中的数据并录入Revman Manager 5.3和Stata 15进行分析。 结果 本研究共计纳入7篇文献,以培养法（Bactec MGIT960）阳性结果作为标准,合并灵敏度80%（95% CI :67%,89%）,合并特异度97%（95% CI :91%,99%）,诊断优势比 DOR =124,95% CI =（27,569）,SROC曲线下AUC面积为0.96（95% CI :0.94,0.97）。以临床诊断（包括培养阳性及培养阴性）作为阳性标准,合并灵敏度为69%（95% CI :29%,93%）,合并特异度为86%（95% CI :65%,95%）。 结论 RNA恒温扩增实时检测技术检测支气管肺泡灌洗液对于痰涂阴性的肺结核患者具有一定的的快速诊断价值,在临床应用上有较好的前景。     

结核感染T细胞酶联免疫斑点试验在肺结核及肺外结核诊断中的价值

目的探讨结核感染T细胞酶联免疫斑点试验(T-SPOT.TB)在肺结核及肺外结核诊断中的价值。方法 T-SPOT.TB检测55例活动性肺结核患者、30例肺外结核患者、14例肺部病变非肺结核者、9例肺外非结核性疾病者和12例健康体检者外周血结核抗原特异性的分泌IFN-γ的T细胞频率。结果肺结核组T-SPOT.TB阳性率明显高于肺部病变非肺结核组和健康对照组,肺外结核组阳性率明显高于肺部病变非肺结核组和健康对照组,差异有统计学意义(P0.01)。T-SPOT.TB检测对菌阳肺结核、菌阴肺结核和肺外结核的诊断敏感度分别为88.57%、80.00%和93.33%,诊断特异度为85.71%、85.71%和88.89%,阳性预测值分别为93.94%、88.89%和96.55%,阴性预测值为75.00%、75.00%和80.00%。菌阴肺结核组与菌阳肺结核组之间比较,T-SPOT.TB诊断效能差异无统计学意义(P0.05);肺外结核组与菌阴肺结核和菌阳肺结核之间比较,T-SPOT.TB诊断效能差异均无统计学意义(P0.05)。结论 T-SPOT.TB诊断结核感染方面具有良好的敏感性和特异性,在肺外结核和菌阴肺结核的诊断与鉴别诊断方面有重要临床应用价值。     

体检和钼靶X线与超声对151例乳腺肿块检查结果比较分析

目的:比较分析体检、钼靶X线与超声诊断乳腺肿块的价值、优势和不足之处。方法:对151例经手术或穿刺明确病理结果的乳腺肿块进行回顾性研究,比较体检、钼靶X线与超声三种检查方法的诊断符合情况。结果:在151例乳腺肿块中,囊肿48例,体检、X线与超声的诊断准确率分别为68.75%、72.92%和93.75%;纤维腺瘤有65例,三者的诊断准确率分别为69.23%、72.31%和95.38%;增生结节27例,三者的诊断准确率分别为48.15%、48.15%和55.56%;乳腺癌11例,三者的诊断准确率为分别72.73%、72.73%和81.82%。结论:体检、钼靶X线与超声是最常用的检查乳腺肿块的方法,在鉴别肿块性质时,三者各有优势和不足之处。但对于良性肿块的诊断,超声更敏感;对于不典型肿块和乳腺癌,主张三者结合综合考虑。     

荧光定量PCR对血及痰培养物中结核分枝杆菌DNA的检测

目的 评价荧光定量PCR(FQ-PCR)检测血及痰培养物中结核分枝杆菌的临床价值.方法 81例临床诊断为结核病的痰涂片阴性患者中,单纯肺结核40例(肺结核组),合并HIV感染的患者41例(合并感染组).患者的血、痰样本分别进行结核分枝杆菌及其L型培养,采用FQ-PCR技术检测血及痰培养液中结核分枝杆菌DNA.结果 肺结核组结核分枝杆菌痰培养FQ-PCR阳性率为54.1%(20/37);血培养阳性率为27.5%(11/40),血培养FQ-PCR阳性率为22.5%(9/40),血培养及其FQ-PCR总阳性率42.5%(17/40).合并感染组10例痰样本培养后2例FQ-PCR阳性;血培养阳性率为7.3%(3/41),血培养FQ-PCR阳性率为17.1%(7/41).合并感染组血培养物FQ-PCR阳性率(17.1%)与肺结核组(22.5%)相比,差异无统计学意义(P>0.05).两组痰与血培养物FQ-PCR总阳性率分别为65.0%(26/40)和22.0%(9/41),差异有统计学意义(χ2=15.305,P<0.01).结论 痰或血培养物FQ-PCR检测有助于提高结核或结核合并艾滋病患者的诊断,同时显著提高了痰涂片阴性肺结核患者的诊断.     

基因芯片技术在雅安市结核分枝杆菌耐药检测中的应用

目的评价基因芯片技术在雅安市结核分枝杆菌耐药检测工作中的应用价值。方法收集2016-06/2018-11雅安市结核病可疑患者痰培养分离株190株进行基因芯片菌种鉴定、耐药检测和比例法药敏试验。以比例法药物敏感试验为金标准,分析基因芯片技术检测结核分枝杆菌对利福平、异烟肼和耐多药诊断的敏感度、特异度、正确指数。采用McNemar检验比较两方法有无差异;采用Kappa检验比较两方法的一致性。两方法有差异的部分样本用基因测序的方法进行比对。结果基因芯片技术进行菌种鉴定,190株分离株中,有186株为结核分枝杆菌,4株为非结核分枝杆菌。以传统比例法药敏试验为金标准,186株结核分枝杆菌对利福平检测的敏感度和特异度为90.0%和96.6%,正确指数为0.866;对异烟肼检测的敏感度和特异度分别为89.5%和99.4%,正确指数为0.889;对耐多药的敏感度和特异度分别75.0%和99.4%,正确指数0.744。两种检测方法比较差异均无统计学意义(McNemar检验,P值分别为0.125、1.000和1.000)。两种检测方法具有较高的一致性(Kappa值分别为0.701、0.910、0.792)。基因芯片技术检测利福平耐药相关rpoB基因突变率最高的位点为531;异烟肼耐药相关katG基因主要突变位点为315。结论基因芯片技术与比例法药敏试验具有较好的一致性,灵敏度高、特异性好,是一种值得推广的耐药结核病诊断方法,在地市级结核病检测中具有重要的应用价值。     

Evaluation of serological tests using A60 antigen for diagnosis of tuberculosis

Identification of acid-fast bacilli (AFB) in sputum or tissue samples is among definite diagnostic methods of tuberculosis. However, this method of diagnosis is restricted by certain limitations. Serologic diagnosis of tuberculosis (TB) has been used for a long time. The aim of this study was to determine the sensitivity and, specificity of Antigen 60 (A60) IgG, IgA, IgM test results in TB diagnosis. Mycobacterial A60-based ELISA was used to measure specific IgA, IgM and IgG antibodies in the sera of 127 adult TB patients (consisted of 74 pulmonary and 53 extra-pulmonary cases), and 95 controls (46 healthy volunteers and 49 patients with various acute or chronic diseases other than tuberculosis). Data from A60 IgG-based ELISA, chest radiography, AFB culture and pathologic evaluation for AFB were obtained .The cutoff value of A60 IgG, IgA and IgM were chosen according to a receiver operating characteristic (ROC) analysis. The sensitivity, specificity and positive likelihood ratio were determined. The mean levels of IgG, IgA and IgM were significantly higher in patients with pulmonary tuberculosis when compared with control groups. Sensitivity of IgG test was 54.3 %, while the specificity was 84.2%. The IgA test showed a sensitivity of 70.1% with a specificity of 80 %. Combination of the IgG and IgA tests showed a total sensitivity of 45.7 % and a specificity of 94.7% and the positive likelihood ratio of 8.62. Chosen cutoff values of IgG, IgA, and IgM sets were 285,265 and 0.9 ELISA units respectively. Our study results showed a good specificity (94.7%) and a reasonable positive likelihood ratio (8.62) of the test when combined IgA and IgG with new cutoff points were considered on diagnosis of tuberculosis in adult patients. Combined use of both IgG and IgA tests results allows an increased accuracy in diagnostic of tuberculosis.     

GenoType MTBDRplus VER 2.0试剂盒检测痰标本中结核分枝杆菌复合群的效果评价

目的评价GenoType? MTBDRplus VER 2.0试剂盒检测痰标本中结核分枝杆菌复合群(MTBC)的检测效能。方法对177例疑似肺结核患者的痰标本同时进行涂片,MGIT液体培养和GenoType? MTBDRplusVER 2.0检测。对培养阳性菌株进行菌种鉴定。使用SPSS 22.0统计软件对结果进行统计学分析,计数资料的一致性比较使用Kappa检验。K值≥0.75认为一致性很好。对MTBC检测的效能比较分析用敏感度、特异度进行效果评估。结果以MGIT液体培养结果为判断标准,GenoType? MTBDRplusVER 2.0检测痰标本中MTBC的敏感度为91.67%,特异度为95.58%,阳性预测值为91.67%,阴性预测值为95.58%,总符合率为94.22%。对两种方法进行一致性Kappa检验,K=0.872(P<0.001)。结论GenoType? MTBDRplus VER 2.0试剂盒检测痰标本中MTBC具有较高的敏感度和特异度,对于结核病的早期诊断有很好的应用价值。     

以多表位融合抗原检测结核分枝杆菌抗体、TB-SA抗体检测和噬菌体裂解法快速检测在结核病诊断中应用价值研究

目的:评价以融合抗原检测结核分枝杆菌抗体、TB-SA抗体检测和噬菌体裂解法快速检测在结核病诊断中应用价值。方法:选择结核组病例134例与非结核组265例按说明书方法进行检测。结果:噬菌体裂解法灵敏度72.39%,特异度91.7%;TB-SA抗体灵敏度75.37%,特异度77.7%;融合抗原检测抗体灵敏度85.8%,特异度92.1%。结论:融合抗原检测抗体与噬菌体裂解法和TB-SA抗体试验相比敏感度更高,值得在各级医疗单位使用。     

应用改良抗酸染色法诊断胸腔炎患者结核杆菌感染的效果分析

目的 分析改良抗酸染色法对结核性胸膜炎患者胸腔积液中结核分歧杆菌的诊断价值。方法 回顾性分析2018年1—12月某医院收治的127例胸腔积液(恶性胸腔积液患者30例,结核性胸腔积液患者97例)患者临床资料,入选者标本均同时采用传统Ziehl-Neelsen抗酸染色、改良抗酸染色法检查。观察比较两种方法的检查结果,以结核菌培养结果作为金标准方法,评价方法的敏感度、特异度、准确率,计算约登指数。结果 传统抗酸染色法敏感度、特异度、准确率分别为:54.64%、83.33%、61.42%,改良抗酸染色法敏感度为80.41%、特异度为96.67%、准确率为84.25%,二者差异有统计学意义(P<0.05);改良抗酸染色法约登指数(0.77)高于传统抗酸染色法(0.38)。结论 改良抗酸染色法有助于提升结核分歧杆菌检出率,较适于为结核性胸膜炎的诊断实验。     

Evaluation and Comparison of Molecular and Conventional Diagnostic Tests for Detecting Tuberculosis in Korea, 2013

ObjectivesA fast and accurate diagnosis is necessary to control and eliminate tuberculosis (TB). In Korea, TB continues to be a serious public health problem. In this study, diagnostic tests on clinical samples from patients suspected to have TB were performed and the sensitivity and specificity of the various techniques were compared. The main objective of the study was to compare various diagnostic tests and evaluate their sensitivity and specificity for detecting tuberculosis.MethodsFrom January 2013 to December 2013, 170,240 clinical samples from patients suspected to have TB were tested with smear microscopy, acid-fast bacilli culture, and real-time polymerase chain reaction (PCR). The test results were compared and data were analyzed.ResultsA total of 8216 cultures tested positive for TB (positive detection rate, 4.8%). The contamination rate in the culture was 0.6% and the isolation rate of nontuberculous mycobacteria was 1.0%. The sensitivity and specificity of smear microscopy were 56.8% and 99.6%, respectively. The concordance rate between the solid and liquid cultures was 92.8%. Mycobacterium isolates were not detected in 0.4% of the cases in the liquid culture, whereas no Mycobacterium isolates were detected in 6.8% of the cases in the solid culture. The sensitivity and specificity of real-time PCR for the solid culture were 97.2% and 72.4%, respectively, whereas the corresponding data for the liquid culture were 93.5% and 97.2%.ConclusionThe study results can be used to improve existing TB diagnosis procedure as well as for comparing the effectiveness of the assay tests used for detecting Mycobacterium tuberculosis isolates.     

结核分枝杆菌7种表位抗原联合检测技术在聚集性疫情处理中的应用

目的:探讨分析在大规模结核疫情筛查中,结核分枝杆菌7种表位抗原联合检测技术的应用价值。方法:使用结核分枝杆菌抗体IgG诊断试剂盒对61份标本进行检测。对12例结核病患者1、9例密切接触者和30例可能有接触者同时进行结核菌素纯蛋白衍生物(PPD)试验、痰抗酸杆菌涂片和培养。结果:结核分枝杆菌抗体(IgG)诊断试剂盒对确认的12例肺结核患者的检测敏感度100%(12/12),健康组30例检测特异性为100%(30/30),19例密切接触者1例抗体阳性。结论:结核分枝杆菌抗体IgG诊断试剂盒具有较好的敏感度和特异度,是大规模筛查和辅助诊断结核病的有效手段。