痰与支气管肺泡灌洗液结核分枝杆菌FQ-PCR检测对涂阴肺结核诊断价值的比较

目的 探讨呼吸道痰标本和支气管肺泡灌洗液(BALF)Mtb-DNA检测对涂阴肺结核的诊断价值。方法 选择福州肺科医院2009年7月至2009年12月胸部影像学疑为肺结核但至少3份痰涂片镜检Mtb阴性,或胸部影像学无法排除肺结核需进一步检查的患者共51例,所有入选患者初诊根据临床症状、体征、胸部影像学结果分为疑似结核组（24例）和待排结核组（27例）。均行抗结核抗体、红细胞沉降率检测及PPD试验,应用实时荧光定量聚合酶链反应（FQ-PCR）定性及定量检测两组患者痰和BALF中的Mtb-DNA。结果 疑似结核组和待排结核组各有22例和2例确诊肺结核,确诊的24例中痰FQ-PCR敏感度为45.8%（11/24）,特异度为96.3%（26/27）,Youden指数42.1%;BALF FQ-PCR的敏感度为75.0%（18/24）,特异度96.3%（26/27）,Youden指数71.3%。结论 FQ-PCR检测BALF Mtb较痰更为敏感,对影像学疑诊但痰涂片阴性的结核患者有很高的实用价值,尤其适用于无痰的患者。     

实时荧光核酸恒温扩增技术在肺结核诊断中的价值分析

目的评价实时荧光核酸恒温扩增技术(SAT-TB)在肺结核诊断中的价值。方法纳入1121例非相同患者痰标本,其中26例为非结核分枝杆菌。570例肺结核为观察组,525例其他肺部疾病患者为对照组。对治疗前的痰标本分别进行抗酸杆菌涂片、培养及鉴定和SAT法检测。阳性检出率的比较使用卡方检验。结果临床诊断肺结核570例,痰培养结核菌阳性率46.5%(265/570),SAT法检测阳性率46.7%(266/570),痰涂片阳性率34.4%(196/570),SAT与涂片法阳性率的差异有统计学意义(χ2=17.83,P0.01)。在涂阴肺结核中,SAT阳性率达19.5%。以培养阳性作为金标准,SAT法诊断结核病的灵敏度为89.4%,特异度96.3%,阳性似然比为24.2,阴性似然比为0.11,一致率是94.6%,阳性预测值是88.4%,阴性预测值为96.6%。在非结核分枝杆菌肺病中SAT检测阳性率为零。结论 SAT-TB法在肺结核的早期诊断中是一种快速、准确、敏感的方法,值得推广。     

结核感染T细胞斑点试验、Xpert Mtb/RIF检测技术在老年肺结核中的诊断价值

目的评价结核感染T细胞斑点试验(T-SPOT)、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(Xpert Mtb/RIF)在老年肺结核中的诊断价值。方法纳入2016年9月至2016年12月在广州市胸科医院住院的老年可疑肺结核患者81例。采集痰标本同时送检抗酸染色涂片、金标准培养并行金标准比例法传统药敏试验、荧光定量PCR和Xpert Mtb/RIF检测,并抽血送检血T-SPOT。以临床最终诊断结果为金标准,分析各种方法诊断老年肺结核的敏感度、特异度。结果 81例老年可疑肺结核患者中,最终诊断肺结核患者44例,非结核肺病患者37例。以临床最终诊断结果为金标准~([10]),血T-SPOT、痰Xpert MTB/RIF、痰荧光定量PCR、痰涂片和痰培养诊断老年肺结核的灵敏度分别为93. 18%、45. 45%、42. 22%、22. 73%和50. 00%;特异度分别为63. 89%、100%、94. 59%、83. 78%和81. 08%。结论血T-SPOT在诊断老年肺结核中有较高的灵敏度,但特异度稍差;痰Xpret MTB/RIF较痰涂片灵敏度提高,特异度达100%。     

四种结核分枝杆菌检测方法的临床应用评价

目的评估涂片、培养、PCR和增菌PCR检测结核分枝杆菌临床应用价值。方法对124例临床确诊的肺结核、可疑结核患者和非结核病人痰标本的涂片、培养、PCR和增菌PCR四种方法的检测结果进行比较。结果涂片、培养、PCR和4及7d的增菌PCR检测31例临床确诊的肺结核病人痰标本阳性率分别为22.5%、32.2%、54.8%、64.5%和87.1%;检测59例临床可疑肺结核病人痰标本阳性率分别为13.6%、18.6%、28.8%、37.3%和52.5%。比较四种方法的阳性检测率有显著性差异(P<0.05)。检测34例非结核病人痰标本,涂片、培养均为阴性,而PCR和增菌PCR均有1例假阳性,假阳性率2.9%。比较PCR与增菌PCR对菌阳和菌阴病人的痰标本阳性检测率,有显著性差异(P<0.05),而两种方法的假阳性率相同。结论增菌PCR检测结核分枝杆菌具有很高的敏感性和特异性,可作为结核病的有效辅助诊断方法之一。     

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诊断艾滋病患者肺结核的临床应用评价

目的:探讨结核分枝杆菌(Mycobacterium tuberculosis,MTB)/利福平耐药实时荧光定量核酸扩增检测(Xpert MTB/RIF)技术诊断艾滋病患者肺结核的应用价值。方法:回顾性分析2017年7月至2019年11月上海市(复旦大学附属)公共卫生临床中心收治的226例疑似肺结核的艾滋病患者的临床资料。分别进行痰涂片荧光染色显微镜下检查、BACTEC MGIT 960液体培养(或罗氏固体培养)和Xpert MTB/RIF检测,并分析在艾滋病患者中Xpert MTB/RIF技术诊断MTB感染和利福平耐药的灵敏度和特异度。结果:226例疑似肺结核患者中,痰培养阳性94例(41.6%),其中MTB阳性51例(54.3%),非结核分枝杆菌(nontuberculosis mycobacteria,NTM)阳性43例(45.7%)。以痰分枝杆菌培养阳性且结核分枝杆菌分泌蛋白64抗原阳性为金标准,Xpert MTB/RIF技术诊断MTB的灵敏度为72.6%[95%可信区间(confidence interval,CI)66.7%~78.4%],特异度为97.1%(95%CI 95.0%~99.3%)。Xpert MTB/RIF技术诊断痰涂片阳性患者MTB的灵敏度为76.7%(95%CI 67.7%~85.8%),特异度为90.0%(95%CI 83.6%~96.5%)。Xpert MTB/RIF技术诊断痰涂片阴性患者MTB的灵敏度为50.0%(95%CI 41.8%~58.2%),特异度为99.3%(95%CI 97.9%~100.0%)。18例患者同时检测利福平表型和基因型耐药,以表型耐药为金标准,Xpert MTB/RIF技术检测利福平基因型耐药的灵敏度为75.0%,特异度为100.0%。结论:艾滋病患者中Xpert MTB/RIF技术诊断肺结核的灵敏度和特异度均较高,且能快速区分MTB和NTM,具有较好的应用价值。     

荧光定量PCR技术在肺结核诊断中的临床应用研究

目的评价荧光定量PCR技术在肺结核病诊断中的应用价值。方法采用荧光定量PCR、金胺O染色荧光镜检和培养法同时检测860例肺结核患者的痰标本,对检测结果进行比较。结果荧光定量PCR检测灵敏度可达10~100 cfu/ml,重复性好,对其他呼吸道病原体检测结果均为阴性。肺结核患者痰标本荧光定量PCR检测阳性率要高于金胺O染色荧光镜检和培养法的阳性率,差异有统计学意义(P0.01)。结论应用荧光定量PCR方法检测结核杆菌,具有特异、灵敏、简便、快速的特点,可作为结核病诊断的方法之一。     

GeneXpert MTB/RIF、T-SPOT检测对老年肺结核的诊断价值

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(GeneXpert MTB/RIF)、结核感染T细胞斑点试验(T-SPOT)在老年肺结核中的诊断价值。方法500例疑似老年肺结核患者痰标本送检GeneXpert MTB/RIF检测,涂片抗酸杆菌染色镜检、分枝杆菌培养、培养阳性菌株比例法药敏试验和菌种鉴定,同时抽血送检血T-SPOT,结核菌素(PPD)试验,与临床诊断结果金标准进行对比,观察各种方法在诊断老年肺结核中的敏感度和特异度。结果最终诊断为肺结核402例,肺部感染53例,慢性阻塞性肺病18例,支气管扩张11例,肺癌9例,间质性肺病7例。血T-SPOT的敏感度为86.6%,特异度为63.3%;PPD试验的敏感度为52.2%,特异度为56.1%;痰GeneXpert MTB/RIF的敏感度为51.7%,特异度为99.0%;痰涂片的敏感度为19.2%,特异度为40.8%;痰培养的敏感度为46.3%,特异度为41.8%。结论老年肺结核诊断方法中,痰GeneXpert MTB/RIF的特异度最高,血T-SPOT的敏感度最高,两者联合检测有利于老年肺结核的早期诊断。     

实时荧光核酸恒温放大检测（SAT）法对肺结核诊断价值的研究

目的 检测实时荧光核酸恒温放大检测（SAT）法对结核病诊断的作用。 方法 收集172例疑似肺结核患者的痰液标本2份/例,1份进行涂涂片金-胺染色,另一份进行罗氏培养基培养与菌种鉴定及SAT法检测结核分枝杆菌,如SAT法和培养结果不符合则使用痰荧光定量PCR法（FQ-PCR)进行复核。 结果 172例入组患者中有156例根据临床及实验室结果确诊为肺结核病。如果以Mtb培养鉴定结果和荧光PCR结果作为判定金标准,SAT检测的敏感度和特异度分别为97.8%（89/91）和97.5%（79/81）。如以临床诊断为标准,SAT诊断结核的敏感度为58.3%（91/156）,特异度为100.0％（16/16）。对于痰菌阳性肺结核患者,SAT 阳性率为84.9%（79/93）;对于痰菌阴性患者,SAT的阳性率为19.0%（12/63）。 结论 SAT法可以作为临床对结核病的辅助诊断方法。     

实时荧光PCR技术在不同类型结核病诊断中的应用价值

目的 分析比较实时荧光PCR与涂片镜检、培养三种方法在结核杆菌检测的差异.方法 对我院疑似结核病患者的临床标本同时进行实时荧光PCR检测、涂片镜检和培养,将三组检测结果进行比较.结果 215例疑似肺结核患者痰标本同时采用实时荧光PCR、涂片镜检和培养三种方法检测,阳性检出率分别为46.5％、32.6％和44.7％.实时荧光PCR法与涂片镜检法的阳性率间比较有统计学意义,实时荧光PCR法与培养法的阳性率同比较无统计学意义.112例疑似其他类型结核病患者的胸腹水、脑脊液、穿刺液和尿液等非痰标本同时采用上述三种方法检测,阳性率分别为18.8％、3.6％和16.1％.统计学意义同上.结论 实时荧光PCR检测技术在临床上对结核病的诊断和鉴别具有较好的应用价值.     

T-SPOT在肺结核中的诊断价值

目的探讨γ干扰素释放结核感染T细胞斑点试验(T-SPOT)在结核病中的诊断价值。方法经临床证实肺结核患者85例,非肺结核患者35例,均给予T-SPOT外周血检测、痰涂片及结核菌素试验(PPD),对三种检查方法进行对比分析。结果 1 T-SPOT检测肺结核灵敏度、特异度、阳性及阴性预测值均明显高于PPD试验(P0.05),但特异度、阳性预测值低于痰涂片检测(P0.05);2 T-SPOT、痰涂片、PPD试验在肺结核中的灵敏度分别为89.41%、63.53%、72.94%(P0.01);3痰涂片检测阳性与阴性肺结核患者T-SPOT阳性率分别为88.89%、90.32%,(P0.05);4 PPD试验强阳性患者T-SPOT阳性率为100%,两者具有高度一致性,明显高于阳性、弱阳性、阴性患者的79.62%、72.72%、86.96%(P0.05)。结论 TSPOT外周血检测对肺结核具有较高的诊断及鉴别诊断价值,减少肺结核的漏诊和误诊,未来可作为肺结核普查方法。     

TaqMan聚合酶链反应技术检测结核分支杆菌DNA及其临？…

目的 探讨ＴａｑＭａｎ聚合酶链反应（ＴａｑＭａｎ－ＰＣＲ）技术在肺结构诊断中的价值。方法 对１６８例活动性肺结核、５７例肺癌患者的痰和外周血及３４－例健康对照外周血,同时应用ＴａｑＭａｎ－ＰＣＲ、ＰＣＲ检测,并与痰涂片法、ＢＡＣＴＥＣ法及改良罗氏培养法结果进行比较。结果 ＴａｑＭａｎ－ＰＣＲ检测痰和外周血总的阳性率分别为５３．０％和６１．３％,显著高于ＰＣＲ、痰涂片法、ＢＡＴＣＴＥＣ法及改良罗氏培     

分子信标荧光定量PCR在肺结核早期诊断及疗效评价中的初步应用

目的探讨分子信标荧光定量PCR在肺结核早期诊断及疗效评价中的应用价值。方法采集2010年3—7月间在彭州市CDC结核病防治所门诊就诊患者的痰标本382份,其中初诊疑似肺结核患者的痰标本305份,肺结核患者随访痰标本77份,用分子信标荧光定量PCR法和抗酸染色法检测Mtb。结果382份痰标本抗酸染色法、分子信标荧光定量PCR法检测总阳性率分别为12.6%(48/382)、42.4%(162/382),分子信标荧光定量PCR法阳性率显著高于抗酸染色法,差异有统计学意义(χ2=74.5,P<0.001)。结论分子信标荧光定量PCR较抗酸染色法具有更高的灵敏度,不仅提高了肺结核早期诊断的检出率,还能间接反映药物疗效,在肺结核防治工作中具有重要意义。     

Yield of gastric lavage and bronchial wash in pulmonary tuberculosis.

OBJECTIVE: To determine the yield of acid-fast bacilli (AFB) in gastric lavage and bronchial washing in adult patients clinically and radiologically suspected of pulmonary tuberculosis but who cannot produce sputum. METHODS: Selected adult patients were admitted to the ward; gastric lavage was done for 3 consecutive days after an overnight fast followed by bronchial wash. Specimens were immediately sent to laboratory for AFB direct smear and culture. RESULTS: The yield of AFB in gastric lavage on direct smear was 16/20 (80%) and 12/20 (60%) in the first and second samples, respectively. When combined, 18/20 (90%) were direct smear positive, while the third sample did not increase the yield. The yield of AFB culture in gastric lavage was 6/20 (30%) in both the first and second samples, while the combined results of the first and second samples were 8/20 (40%). The third sample did not increase the yield. In bronchial wash, AFB direct smear was positive in 18/20 (90%), while culture was positive in 14/20 (70%). CONCLUSION: Gastric lavage and bronchial washing are useful methods for the diagnosis of pulmonary tuberculosis in patients who cannot produce sputum. Two gastric lavage specimens are adequate. On comparison, bronchial wash is superior to gastric lavage in culture, but their yield on direct smear is equal.     

聚合酶链反应对菌阳肺结核治疗的监测

目的探讨痰菌阳性肺结核患者在治疗期及停药２年内痰结核分支杆菌及其ＤＮＡ阴转情况与复发的关系以及聚合酶链反应（ＰＣＲ）对菌阳肺结核患者治疗的监测价值。方法用ＰＣＲ技术、涂片及培养法对８７例菌阳肺结核于治疗期每月检测１次，停药期继续随访２年。结果痰结核分支杆菌ＰＣＲ转阴时间通常比涂片和培养迟１～３个月，痰含菌量越多，ＰＣＲ持续阳性时间越长。８７例中１０例（１１％）ＰＣＲ持续阳性１年以上，其中３例（３０％）分别于停药后８、１２、１６个月时复发，１例ＰＣＲ已转阴病例于１８个月时复发。此４例均有痰菌复阳，胸片示病灶增多而再次接受治疗。结论ＰＣＲ用于临床疗效观察比涂片、培养实用，对估计有可能复发的病例有一定帮助。     

Detection of Mycobacterium tuberculosis in bronchoalveolar lavage from patients with sputum smear-negative pulmonary tuberculosis using a polymerase chain reaction assay

Abstract The objective of this study was to evaluate the utility of a polymerase chain reaction (PCR) assay in detecting Mycobacterium tuberculosis in bronchoalveolar lavage (BAL) specimens of patients suspected of having active pulmonary tuberculosis (TB) but who were sputum smear-negative. Patients undergoing investigation for suspected pulmonary TB at the University Hospital, Kuala Lumpur, and who were sputum smear-negative underwent fibreoptic bronchoscopy and BAL. One portion of each lavage specimen was submitted for smear examination for acid-fast bacilli and mycobacterial culture and the other portion assayed by PCR for the presence of a 562-base pair DNA segment belonging to the insertion sequence IS986, unique to the M. tuberculosis complex. As controls, lavage specimens from patients with other lung lesions were also similarly tested. The PCR assay gave a positivity rate of 80.9% (55 of 68) compared with 8.8% of smear examination and 7.4% of culture for detecting M. tuberculosis in BAL specimens. The assay was positive in two of 45 BAL specimens from 35 control subjects. The PCR assay was more sensitive than smear and culture in detecting M. tuberculosis in BAL specimens of patients with sputum smear-negative pulmonary TB.     

Value of smear and PCR in bronchoalveolar lavage fluid in culture positive pulmonary tuberculosis.

At present, further investigations are needed in patients with suspected pulmonary tuberculosis (TB) and either negative sputum smear or without sputum. The aim of the present study was to analyse the yield of bronchoalveolar lavage fluid (BALF) smear and PCR in patients with confirmed pulmonary TB. Patients with a positive culture for Mycobacterium tuberculosis complex in sputum or BALF were analysed over 5 yrs. In total, 90 out of 230 (39%) patients with culture-positive pulmonary TB had a positive sputum smear, and 120 patients underwent bronchoscopy. BALF smear was positive in 56 (47%), BALF PCR in 93 (78%) patients, and BALF smear and/or PCR was positive in 83%. In total, 71 patients who underwent bronchoscopy and had complete clinical records were further analysed. BALF (smear or Mycobacterium tuberculosis complex-PCR) allowed a rapid diagnosis in 10 (59%) out of 17 patients who had a negative sputum smear, and 49 (91%) out of 54 patients without sputum production. Of these 71 patients, 12 (17%) were only culture positive. Rapid diagnosis of pulmonary TB by smear and/or PCR was made in 190 out of 210 patients (90%) in sputum or BALF. In conclusion, combined use of bronchoalveolar lavage fluid smear and Mycobacterium tuberculosis complex-PCR has a good diagnostic yield in patients with sputum smear-negative tuberculosis or without sputum production.     

痰涂片阳性结核病患者的涂片与培养结果分析

目的 研究痰涂片阳性(简称涂阳)结核病患者不同标本涂片与罗氏培养阳性率的差别,探讨痰涂片和痰培养的标本选择标准.方法 回顾性分析2007年全国结核病耐药性基线调查纳入的结核病涂阳患者中3813例的痰涂片和培养结果,比较即时痰、夜痰和晨痰的涂片和培养阳性率,以及每300个视野中检测出1～2条抗酸杆菌的标本培养阳性率.两两比较采用χ2检验.结果 即时痰、夜间痰和晨痰的涂片阳性率分别为75.2%(2868/3813)、87.9%(3350/3813)和89.4%(3408/3813);同一病例的3份痰涂片均为阳性者占64.5%(2460/3813),2份涂片为阳性者占23.4%(893/3813),仅1份痰涂片为阳性者占12.1%(460/3813);即时痰加晨痰的涂片累积阳性率是3份痰涂片阳性率的96.9%(3695/3813),即时痰、夜间痰和晨痰的培养阳性率分别为97.8%(1654/1691)、96.7%(3006/3107)和97.2%(2473/2543);涂阳病例2份痰比1份痰的培养阳性率仅提高1.7%～2.8%;300个视野中检出1～2条抗酸杆菌的痰标本中90%(98/109)为痰培养阳性.结论 如采集即时痰和晨痰用于初诊者诊断,可将涂片检查3份痰标本改为2份;按照涂阳标本优先培养的原则,初诊涂阳患者培养可以由2份标本改为1份标本;涂片检查结果为每300个视野1～2条抗酸杆菌时具有一定诊断价值.

Abstract：

Objective To study the positive rate of smear and culture in different sputa from smearpositive patients with tuberculosis,and to discuss the criteria of sputum selection for smear and culture.Methods The results of smear and culture of sputum samples from 3813 cases during tubercuiosis drugresistance baseline survey in 2007 were retrospectively analyzed.The positive rates of smear and culture of spot sputum,night sputum and morning sputum were compared,and the culture positive rate of sputum samples with 1-2 acid fast bacilli(AFB)/300 fields was summarized.The χ2 teat was used for comparison.Results The smear positive rate of spot sputum,night sputum,and morning sputum wag 75.2%(2868/3813),87.9%(3350/3813),and 89.4%(3408/3813)respectively.The percentage of cases which were positive for all the 3 sputum samples was 64.5%(2460/3813),while that for 2 samples and 1 sample was 23.4%(893/3813)and 12.1%(460/3813)respectively.The cumulative smear positive rate of spot sputum plus morning sputum reached 96.9%(3695/3813)of the rate combined from the 3 sputum samples.The culture positive rate of spot sputum,night sputum,and morning sputum was 97.8%(1654/1691),96.7%(3006/3107)and 97.2%(2473/2543)respectively.For the smear positive cases,the positive rate of 2 sputum culture increased only 1.7%-2.8%compared with that of 1 sputum culture.The culture positive rate of samples with a smear result of 1-2 AFB/300 fields was 90%(98/109).Conclusions When spot sputum and morning sputum are collected for the diagnosis,the humber needed for smear can be decreased from 3 to 2.For smear-positive cases,if sputum is selected for culture according to the smear result,the number needed for culture can be decreased from 2 to 1.A smear result of 1-2 AFB/300 fields is of considerable diagnostic value for tuberculosis.     

Diagnostic value of bronchoscopy in diagnosis of pulmonary tuberculosis: bronchial aspirate, bronchial washing and transbronchial lung biopsy

The diagnosis of pulmonary tuberculosis is confirmed by the detection of Mycobacterium tuberculosis in sputum. Bronchoscopy has been used for diagnosis of various pulmonary diseases. The value of bronchoscopy such as bronchial aspirate, bronchial washing and transbronchial lung biopsy in diagnosis of pulmonary tuberculosis was evaluated, and the results were as follows: 1) One hundred ninety cases were investigated bronchoscopically due to suspicion of pulmonary tuberculosis with sputum negative smear and 92 cases were confirmed to be pulmonary tuberculosis. 2) Out of 91 cases examined by bronchial aspirate and 46 cases by bronchial washing, smear positivity was 20.9% and 23.9% and culture positivity was 58.2% and 84.8%, respectively. Transbronchial lung biopsy showed positive findings of tuberculosis in 75.8% out of 33 specimens. 3) Out of 88 sputa taken before bronchoscopy and 50 sputa after bronchoscopy, smear positivity was 0% and 12%, and culture positivity was 54.5% and 40% respectively. Gastric lavage culture positivity was 29.4% in 17 cases examined. 4) Diagnosis of tuberculosis was made rapidly in 28 cases (30.4%) by smear positive results of bronchial aspirate, bronchial washing and sputa after bronchoscopy, and relatively rapidly in 20 cases (21.7%) by transbronchial lung biopsy.