肺泡灌洗液不同检测方法在肺结核诊断中的临床价值

目的比较灌洗液标本的荧光定量PCR、纤支镜灌洗液刷片抗酸染色和结核分枝杆菌快速培养的灵敏度和特异性对于肺结核的临床诊断应用价值。方法选取安徽省胸科医院2016年1月-2016年12月临床确诊的649例肺结核患者、1328例肺炎、肺脓肿等其它呼吸系统疾病共1977例患者送检的纤维支气管镜灌洗液标本,同时进行灌洗液荧光定量PCR、纤支镜灌洗液结核分枝杆菌快速培养和灌洗液刷片抗酸染色3种方法的检查,比较其灵敏度和特异性。结果灌洗液荧光定量PCR、灌洗液结核分枝杆菌快速培养和灌洗液刷片抗酸染色的灵敏性分别为80. 3%,66. 3%和38. 1%,特异性分别为97. 7%,97. 4%和97. 1%。结论纤维支气管镜肺泡灌洗液的荧光定量PCR对于辅助肺结核的快速诊断较其他方法有较明显的优势,对肺结核的诊断有重要意义。     

应用免疫磁珠分离-改良罗氏培养技术检测痰结核分枝杆菌的研究

目的通过免疫磁珠分离技术(immunomagnetic beads separation techniques,IMBS)富集痰液中的结核分枝杆菌,提高痰液培养的阳性率,以期用于结核病的快速诊断。方法制备兔抗结核杆菌IgG抗体,并将其结合于免疫磁珠表面。采集71例确诊结核病患者的痰液标本,通过免疫磁珠分离技术捕获、富集吸附结核分枝杆菌后用改良罗氏培养基培养,并与传统改良罗氏(L-J)培养法和痰涂片抗酸染色法作比较。结果 71例结核病患者痰涂片抗酸染色检查阳性26例,阳性率36.6%;传统改良L-J培养阳性34例,阳性率47.9%;免疫磁珠分离技术捕获、富集结核分枝杆菌后进行改良L-J培养阳性48例,阳性率67.6%。三者阳性率差异有统计学意义(P<0.05),改良L-J培养和痰涂片检查阳性率差异无统计学意义(P>0.05)。结论采用免疫磁珠分离技术捕获、富集痰液中的结核分枝杆菌后进行培养,可显著提高痰培养的阳性率,该方法可用于结核病的快速诊断。     

抗酸染色全自动显微扫描识别系统的临床应用评价

目的 评价抗酸染色全自动显微扫描系统(AFB-AS)检测肺结核患者痰液标本应用价值。方法 搜集2019年1月至8月于北京结核病控制研究所确诊的462例肺结核患者作为研究对象,每例患者留取3份初诊痰液标本,共计1386份。所有标本均完成萋-尼抗酸染色,采用双盲法使用传统抗酸杆菌染色镜检和AFB-AS检查;每例患者选取2份痰液标本进行分枝杆菌分离培养,共计完成924份;其中1份同时进行GeneXpert MTB/RIF(简称“Xpert”)检测,共计完成462份。比较不同方法检测肺结核患者痰标本结果;以分枝杆菌分离培养结果为参照,分析传统抗酸杆菌染色镜检和AFB-AS检测肺结核患者痰标本的效能;分析传统抗酸杆菌染色镜检和AFB-AS检测均为阳性结果的等级相关性。结果 各方法检测肺结核患者痰标本阳性率从高到低依次为Xpert(69.26%,320/462)、分枝杆菌分离培养(43.61%,403/924)、AFB-AS(43.22%,599/1386)、传统抗酸杆菌染色镜检(38.24%,530/1386)。AFB-AS检测痰标本阳性率明显高于传统抗酸杆菌染色镜检,差异有统计学意义(χ²=67.015,P<0.01)。AFB-AS与传统抗酸杆菌染色镜检判读阳性结果等级的相关系数为0.954。以分枝杆菌分离培养结果为参照,AFB-AS检查肺结核患者痰标本敏感度为94.29%(380/403)、特异度为90.40%(471/521)、阳性预测值为88.37%(380/430)、阴性预测值为95.34%(471/494),Kappa值为0.841。结论 AFB-AS检测肺结核患者痰液标本具有较好的应用价值,检验性能能够满足实施萋-尼抗酸染色查找抗酸杆菌项目的要求。     

荧光染色法在结核分枝杆菌检测中的应用

目的 观察荧光染色法在肺结核患者痰标本中结核分枝杆菌的阳性率检测中的临床应用价值.方法 250例肺结核患者,收集深咯痰标本,每份痰标本涂抹2张玻片,分别采用荧光染色法（荧光染色组）和抗酸染色法（抗酸染色组）检测结核分枝杆菌,比较两组阳性率.结果 荧光染色组检出结核分枝杆菌阳性109例,阳性率为43.60％,其中（＋）、（＋＋）、（＋＋＋）、（＋＋＋＋）阳性率分别占13.20％、11.60％、8.40％、10.40％;抗酸染色组阳性86例,阳性率为34.40％,其中（＋）、（＋＋）、（＋＋＋）、（＋＋＋＋）分别占6.80％、8.80％、8.00％、10.80％.两组总阳性率及（＋）阳性率比较P均＜0.01.与抗酸染色组比较,荧光染色组检测痰中结核杆菌的灵敏度、特异性均较高（P均＜0.05）.结论 荧光染色法检测肺结核患者痰标本中结核分枝杆菌的阳性率高于抗酸染色法,且差异主要体现在含菌量少的痰标本上,这对于那些病变活动性轻,排菌量较少的肺结核的诊断具有重要意义.     

荧光定量PCR测定支气管肺泡灌洗液中TbDNA对涂阴肺结核的诊断价值

目的探讨纤支镜肺泡灌洗液中结核分枝杆菌DNA在涂阴肺结核诊断中的价值。方法应用荧光定量聚合酶链反应（FQ-PCR）技术对300例涂阴肺结核,178例菌阳肺结核及119例肺炎或肺癌患者的纤支镜肺泡灌洗液标本进行结核分枝杆菌DNA检测。结果3种病因肺泡灌洗液结核分枝杆菌DNA阳性检出率分别为：74.3%（223/300）、94.9%（169/178）、17.6%（21/119）。结论荧光定量聚合酶链反应检测肺泡灌洗液结核分枝杆菌DNA,用于肺结核诊断可提高肺结核诊断的敏感性和准确性,明显优于痰抗酸染色,亦较痰结核菌培养诊断灵敏、快速,可作为肺结核诊断较可靠指标。     

液基夹层杯技术检测抗酸杆菌应用性研究

目的评价液基夹层杯技术检测抗酸杆菌的应用价值。方法收集1 190例肺结核可疑症状者的642份晨痰标本和1 140份即时痰标本,每份痰标本均采用液基夹层杯技术、直接涂片查找抗酸杆菌技术和罗氏结核分枝杆菌培养技术进行检测,比较直涂法、液基法与培养法对痰标本的阳性检出率,并在以培养法为金标准的前提下考察液基法、直涂法的灵敏度和特异度,同时评价晨痰和即时痰、1次痰和2次痰对肺结核涂阳病人发现的差异。结果液基法、直涂法和培养法对642份晨痰标本的阳性检出率分别为23.5%、12.7%和26.6%,对1 140份即时痰的阳性检出率分别为18.1%、9.6%和23.3%。以培养法作为金标准,液基法的灵敏度和特异度分别为68.9%和95.8%,并与直涂法相对应的指标差异有统计学意义(P0.05)。液基法检测表明:642份晨痰阳性检出率显著高于1 140份即时痰阳性检出率(P0.05)。结论液基法能够提高痰标本的灵敏度,特异度较高,操作标准化,且所需设备简单,便于基层开展,是一种可能具有推广价值的检测方法。     

痰结核分枝杆菌DNA检测诊断肺结核的临床意义

目的评估痰结核分枝杆菌DNA检测诊断肺结核的可行性及临床意义。方法回顾性分析2016年5月—12月在我科住院的经痰结核分枝杆菌培养确诊的136例肺结核患者的临床资料,比较痰涂片找抗酸杆菌和痰结核分枝杆菌DNA检测2种方法的检出率。结果 136例确诊的肺结核患者中,痰涂片阳性的有64例(47.06%),痰结核分枝杆菌DNA检测阳性的88例(64.71%),痰结核分枝杆菌DNA检测阳性率明显高于痰涂片找抗酸杆菌(P0.05)。72例痰涂片阴性患者中痰结核分枝杆菌DNA检测阳性29例,占痰涂片阴性肺结核患者比例的40.28%。结论痰结核分枝杆菌DNA检测优于痰涂片找抗酸杆菌,能够辅助涂阳肺结核的诊断,并提高涂阴肺结核的诊断水平,是一种快速、敏感的病原学诊断方法。     

荧光定量PCR测定支气管肺泡灌洗液中TbDNA诊断肺结核的临床价值

目的探讨纤支镜肺泡灌洗液中结核分枝杆菌DNA在肺结核诊断中的价值。方法对187例初治肺结核患者及41例肺炎患者分别应用荧光定量聚合酶链反应(FQ-PCR)技术检测结核分枝杆菌DNA、肺泡灌洗液结核菌培养、纤维支气管镜刷片抗酸染色。结果肺结核患者肺泡灌洗液结核分枝杆菌DNA阳性率为:78.6%、肺泡灌洗液结核菌培养阳性率为28.8%、刷片抗酸染色阳性率为21.9%。肺炎患者肺泡灌洗液结核分枝杆菌DNA、肺泡灌洗液结核菌培养、刷片抗酸染色均阴性。结论荧光定量聚合酶链反应检测肺泡灌洗液结核分枝杆菌DNA,特异性高,用于肺结核诊断可提高肺结核诊断的敏感性和准确性,明显优于纤维支气管镜刷片抗酸染色,亦较肺泡灌洗液结核菌培养诊断灵敏、快速,可作为肺结核诊断较可靠指标。     

BACTEC MGIT 960和BACT/ALERT 3D系统快速培养和鉴定痰中抗酸杆菌

目的对抗酸染色阳性痰行分枝杆菌培养和鉴定。方法采用萋一尼氏抗酸染色法对临床诊断肺结核患者的晨痰涂片直接镜检,抗酸染色阳性痰经BACTEC960和BACT/ALERT3D系统进行分枝杆菌培养,分别经对硝基苯甲酸（PNB）和噻吩-2-羧基肼（TCH）培养基生长试验行分枝杆菌菌群和结核分枝杆菌复合群菌种鉴定。结果抗酸染色阳性痰标本的分枝杆菌培养阳性率100％,大多数为结核分枝杆菌（9／10）,少数为非结核分枝杆菌（1／10）,最快6天即可报告分枝杆菌阳性培养。结论BACTEC 960和BACT／ALERT3D系统具有快速培养分枝杆菌作用,抗酸染色阳性痰有必要行分枝杆菌培养和鉴定,有利于肺结核与非结核分枝杆菌病的鉴别诊断、结核分枝杆菌菌种鉴定和抗结核药物敏感性试验。     

痰液标本结核分支杆菌RNA恒温扩增检测及临床应用

目的探讨实时荧光核酸恒温扩增检测法（SAT）在肺结核患者实验诊断中的应用。方法设计特异性引物和探针,建立并优化SAT检测体系,对131例临床确诊并接受治疗的肺结核患者、73例非结核性肺部感染者痰液标本中的结核分支杆菌（TB）RNA进行检测,并与PCR法检测TB DNA,痰培养法、痰涂片抗酸染色法检测TB的结果进行比较。结果 131例肺结核患者痰液标本中,SAT对TB检出率为52.67%,高于痰培养法（39.69%,P=0.035）和痰涂片抗酸染色法（35.11%,P=0.004）;以PCR法为参考标准,SAT的灵敏度、特异度、阳性预测值、阴性预测值依次是95.38%、90.91%、91.18%、95.24%。结论 SAT检测痰液标本中TB RNA灵敏度高,特异性强,操作简便、快速,具有临床推广应用价值。     

Evaluation of the Idaho Technology LightCycler PCR for the direct detection of Mycobacterium tuberculosis in respiratory specimens.

SETTING: The rapid detection of Mycobacterium tuberculosis (TB) in clinical samples is an important goal. The LightCycler heralds an advance in thermal cycle technology combining rapid cycle DNA amplification with fluorimetry, eliminating the need to perform amplification and product analysis separately. OBJECTIVES: To evaluate the LightCycler for direct detection of M. tuberculosis complex in respiratory specimens. To evaluate a DNA extraction method based on Chelex 100 resin, heating and ultrasonication for the prevention of endogenous inhibitions in respiratory samples. DESIGN: DNA was extracted from sputum samples using the Chelex method and polymerase chain reaction (PCR) for TB performed with the LightCycler. RESULTS: For 88 sputum samples positive by microscopy and culture for M. tuberculosis, 95% were PCR-positive. None of the five sputum samples that were smear-negative but culture-positive for M. tuberculosis, the 79 culture-negative sputum samples and the 29 sputum samples that were culture-positive for mycobacteria other than TB yielded positive PCR results. PCR inhibitors were not detected in any of the samples. CONCLUSION: The LightCycler proved a simple, reproducible and rapid system, reducing the time to result from weeks (culture) or days (conventional PCR) to hours. The Chelex 100 resin method produced good results for the smear-positive specimens. However, a larger study is required to determine the efficacy of the method with smear-negative specimens and for specimens known to contain endogenous inhibitors.     

Tuberculosis in east timorese refugees: implications for health care needs in East Timor.

SETTING: East Timorese refugees evacuated to Darwin, Australia, September 1999. OBJECTIVE: Presentation of the process and results of tuberculosis (TB) screening in a previously unscreened refugee population. DESIGN: Screening for TB by clinical examination (all persons) and chest X-ray (CXR) (persons over 12 years of age and those of any age with respiratory symptoms) and sputum microscopy and mycobacterial culture (abnormal CXR). RESULTS: Seventy-six patients were diagnosed with TB (38 culture-positive for Mycobacterium tuberculosis, including 11 sputum smear-positive). Of 89 positive mycobacterial cultures, 51 were non-tuberculous mycobacteria (NTM). Of the M. tuberculosis isolates, 82.2% were fully sensitive, 17.2% were resistant to isoniazid and 8.6% were resistant to isoniazid and streptomycin. Fifty-three consecutively diagnosed patients with TB were HIV-negative. The TB burden in this population was very high (point prevalence of 542/100,000 for smear-positive and 2,060/100,000 for culture-positive cases). Rates of culture for NTM were also high. Information from this study assisted the implementation of a National TB Control Programme for East Timor in February 2000. CONCLUSION: The challenges for public health authorities in East Timor to provide a successful TB control programme are enormous. The apparently low prevalence of drug resistance and HIV co-infection in the population is encouraging.     

Etiology of pulmonary infections in predominantly HIV-infected adults with suspected tuberculosis, Botswana.

SETTING: In countries with high HIV rates, diagnosis of lower respiratory disease etiology is both challenging and clinically important. OBJECTIVE: To determine the etiology of lower respiratory tract disease among persons with suspected tuberculosis (TB) and abnormal chest X-rays in a setting with very high HIV seroprevalence. DESIGN: Cross-sectional prevalence data from a prospective cohort of predominantly hospitalized adults with suspected TB in Botswana, January-December 1997. RESULTS: Of 229 patients, 86% were HIV-positive and 71% had a pathogen identified. TB was confirmed in 52%, 17% had acute mycoplasma pneumonia, 3% had Pneumocystis carinii, 27% grew a bacterial pathogen from sputum and 8% from blood. Ninety-four per cent of TB diagnoses were made through expectorated sputum and only 5% of TB cases were diagnosed by sputum induction alone. Polymerase chain reaction (PCR) for Mycobacterium tuberculosis had positive and negative predictive values of 94% and 59%, respectively. Male sex, cough < 2 weeks, and tuberculin skin test > or = 5 mm were independently associated with culture-positive TB among persons with negative acid-fast bacilli smears. Co-infection with two or more pathogens occurred in 25%. CONCLUSIONS: Mycoplasma pneumoniae infection was quite common despite clinical suspicion of TB, and sputum induction and PCR did not significantly improve our ability to diagnose TB, although clinical presentation had some predictive value.     

Novel method for sputum induction using the Lung Flute in patients with suspected pulmonary tuberculosis

Background and objective:   The Lung Flute is a small self-powered audio device that generates sound waves, which vibrate in tracheobronchial secretions. This was a preliminary trial to evaluate the usefulness of the Lung Flute for sputum sampling in patients suspected of pulmonary tuberculosis (TB).
Methods:   Thirty-four patients who were not expectorating sputum, but for whom sputum examination was required for the differential diagnosis of TB or other diseases, were enrolled in the study. Patients were instructed to blow out fast and hard through the Lung Flute and to repeat this for a total 20 sets of two blows each.
Results:   Using the Lung Flute, sputum samples were collected within 10 or 20 min from 30 of 34 patients (88%). The device permitted a rapid diagnosis of TB in seven of 15 confirmed TB cases. In three patients acid-fast bacillus smears were positive. In four patients acid-fast bacillus smears were negative, but PCR tests for TB were positive. Hyperventilation-related symptoms occurred in three patients.
Conclusions:   The application of the Lung Flute may represent a promising technique for the rapid diagnosis of pulmonary TB.     

西安市结核病的病原学调查

目的调查西安市结核病的分枝杆菌菌种类型及其分布。方法采用抽样调查方法收集结核病患者的痰标本及其背景资料,用痰涂片、痰培养以及PCR方法检测,用鉴别培养基对分离菌株进行菌种鉴定。结果共收集574例拟诊结核病病例,实验室检测1602份痰标本。采用痰涂片、痰培养和PCR3种方法联合应用,阳性者260例,分枝杆菌检测阳性率45.30。137例分枝杆菌培养阳性者中,结核分枝杆菌复合体占73.72,其中结核分枝杆菌和牛分枝杆菌分别为67.88和5.84;11.68为非结核分枝杆菌,14.60为混合感染,其中11.68为结核分枝杆菌与牛分枝杆菌,2.92为结核分枝杆菌与非结核分枝杆菌混合感染。结论西安市结核病的致病菌复杂,包括结核分枝杆菌、牛分枝杆菌和非结核分枝杆菌,尤其是非结核分枝杆菌混合感染应予以高度重视。     

结核抗体lgG/lgM检测在肺结核和肺外结核中的诊断价值

目的 探讨异种血清抗体检测技术在结核病诊断中的应用价值。方法 采用IgG/IgM抗体试剂盒分别检测102例结核病患者(包括73例肺结核和29例肺外结核)、223例其他肺部疾病患者和100例对照者结核感染情况,以临床诊断为标准评价该方法的敏感度和特异性,同时分别与痰菌培养及痰涂片平行检验的结果作比较,统计学分析采用χ²检验。结果 结核抗体IgG/IgM检测结核病患者的敏感度为74.51%、特异度为91.64%。结核抗体IgG/IgM检测肺结核和肺外结核的敏感度分别为82.19%、55.17%,肺内和肺外结核的敏感度差异有统计学意义(P＜0.05),结核抗体lgG/IgM检测结核患者阳性检出率明显高于痰培养法和痰涂片法(P<0.05)。102例结核病患者年龄段分组分析,少年组和老年组检出率分别为58.33%、36%,远低于青年组和中年组的96.15%和89.74%。不同年龄组间进行卡方比较分析显示,P＜0.05,差异有统计学意义。425例标本中,共发现8例非结核分枝杆菌,其中6例胞内分枝杆菌, 2例脓肿分枝杆菌,lgG/lgM抗体检测均为阴性。结论 IgG/IgM血清抗体检测肺内、外结核具有快速方便、经济和较高的敏感度,适合用于临床结核筛查。     

Diagnostic yield of repeated smear microscopy examinations among patients suspected of pulmonary TB in Shandong province of China.

OBJECTIVES: To analyse the yield of five repeated smear microscopy examinations for the diagnosis of smear-positive pulmonary tuberculosis (TB). METHODS: Patients with respiratory symptoms and abnormal chest X-rays provided five spontaneous sputum samples for acid-fast bacilli (AFB) smear microscopy in one of nine county laboratories. RESULTS: Of 9302 patients with respiratory symptoms and abnormal X-rays, 6437 (69%) had at least one smear-positive sputum. Of these, 84.5% were diagnosed on the first smear, 96.7% on the first two smears, and 99.9% on the first three sputum smears. The fourth and fifth sputum smears yielded only seven additional cases (0.1%). CONCLUSIONS: Smear microscopy examination of two spontaneous sputum specimens is the most efficient, and three sputum smear examinations provide a diagnosis in almost all cases.     

Diagnostic value of gastric aspirate smear and polymerase chain reaction in smear-negative pulmonary tuberculosis

OBJECTIVE: The aim of this study was to determine the validity of acid-fast bacilli (AFB) smear and polymerase chain reaction (PCR) from gastric aspirates for the diagnosis of smear-negative pulmonary tuberculosis. METHODOLOGY: A cross-sectional study was conducted in a university hospital. One hundred and nine patients with suspected pulmonary tuberculosis in whom either sputum smears were negative or who were not producing sputum were recruited to the study. All patients underwent gastric aspiration after an overnight fast followed by standard fibreoptic bronchoscopy. Specimens were subjected to AFB smear, culture, and pathological examination. PCR was performed on culture filtrate after 1 week of incubation. RESULTS: Eight patients did not complete the follow-up schedule. Of the 101 patients with final outcomes, a diagnosis of pulmonary tuberculosis from microbiological evidence was established in 54 patients. The gastric aspirate smear, PCR, or either one of them was positive in 34, 30, and 39 tuberculosis patients, respectively. There were 13 false positive smears from 47 non-tuberculosis patients, with five resulting from non-tuberculous mycobacteria (NTM). The PCR was falsely positive in eight patients, five of whom had previous histories of tuberculosis. The overall sensitivity, specificity, positive predictive value, and negative predictive value of gastric aspirate examination by combined smear and PCR were 72, 58, 66, and 64%, respectively. CONCLUSIONS: Gastric aspiration is a useful tool for the diagnosis of smear-negative pulmonary tuberculosis warranting institution of antituberculosis treatment. Interpretation of the results should be cautious in those who have had tuberculosis in the past or who have been at risk for acquisition of NTM.     

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。