聚合酶链反应在诊断结核性胸腔积液中的应用

应用聚合酶链反应（ＰＣＲ）技术对６２例胸水进行结核菌ＤＮＡ检则，并同时与胸水涂片及培养结果进行比较。结果显示：３４例结核性胸水涂片抗酸染色、结核菌培养和ｐＣＲ检测阳性率分别为５．９％、８．８％和５２．９％，后者显著高于前二者（Ｐ均＜０．０１）。２８例非结核性胸水涂片和培养结果均呈阴性，但ＰＣＲ有４例（１４．３％）出现阳性。结果表明ＰＣＲ直接检测胸水中结核菌显示出快速、敏感和高效等优点。同时还对影响ＰＣＲ检测结核菌的某些因素作了分析。     

PCR检测结核菌的应用评价

应用ＰＣＲ方法检测了３８例结核性胸膜炎和１２例结脑患者的胸水及脑脊液中的结核菌，阳性率分别为５５．３％和８３．３％，而传统检查方法的阳性率分别为７．６％和８．３％，两种方法比较，有显著性差异。表明ＰＣＲ方法用于检测结核菌具有敏感和特异性强的优点。     

聚合酶链反应早期诊断结核性胸膜炎的临床价值

聚合酶链反应早期诊断结核性胸膜炎的临床价值肖志坚，崔秀琴，肖志敏，张菊萍，杨素敏为提高对结核性胸膜炎（结胸）的早期确诊率，对４８例结胸患者进行了胸水、血清中ＰＣＲ－ＴＢ－ＤＮＡ（用聚合酶链反应技术检测结核菌特异性ＤＮＡ片段）和结核抗体检测，胸水涂片结...     

套式聚合酶链反应快速诊断结核性心包炎

快速诊断结核性心包炎以便及早抗痨治疗防止缩窄性心包炎的发生。应用套式聚合酶链反应（Ｎｅｓｔｅｄ－ ＰＣＲ） 、直接涂片、培养对８３ 例心包积液标本进行结核菌检测。结果：结核性心包积液Ｎｅｓｔｅｄ － ＰＣＲ阳性率为７２ ．５ ％ 、培养阳性率为１９ ．６ ％ 、涂片镜检阳性率为２３ ．５ ％ 。非结核性心包积液无１ 例阳性。Ｎｅｓｔｅｄ－ ＰＣＲ特异性好、敏感性强并可能避免一般聚合酶链反应（ＰＣＲ） 所出现的假阳性     

聚合酶链反应诊断结核性腹膜炎的评价

目的评价聚合酶链反应（ＰＣＲ）方法对结核性腹膜炎的诊断价值。方法用ＰＣＲ技术检测３０例结核腹水中结核分支杆菌ＤＮＡ，并与腹水涂片抗酸染色及酶联免疫吸附试验（ＥＬＩＳＡ）检测腹水中抗ＰＰＤ抗体进行比较。结果ＰＣＲ的阳性率为６０％，特异性９４．４％；ＥＬＩＳＡ法阳性率６３．３％，特异性７２．２％；涂片镜检均为阴性。结论ＰＣＲ在诊断结核性腹膜炎具有较高的敏感性和特异性，优于ＥＬＩＳＡ法及涂片镜检，如与ＥＬＩＳＡ技术结合可进一步提高检测的敏感性和特异性。     

应用PCR,BA—ELISA早期诊断结核性胸膜炎的临床意义

应用ＰＣＲ检测６４例胸水中结核菌基因组ＤＮＡ，并与ＢＡ－ＥＬＩＳＡ检测胸水中抗ＰＰＤ抗体及细菌学检查相比较。结果：三种方法的敏感性和特异性分别为５６．０％，１００％；６４．０％，８６．７％，１１．０％和１００％。若以ＰＣＲ，ＢＡ－ＥＬＩＳＡ任一阳性作为判断标准则敏感明显提高至８９．０％。认为，ＰＣＲ与ＢＡ－ＥＬＩＳＡ诊断结核性胸膜炎具有较高的敏感性，但前者更具早期和特异性并对疾病的疗效和预后有判断     

应用PCR、BA－ELISA早期诊断结核性胸膜炎的临床意义

应用ＰＣＲ检测６４例胸水中结核菌基因组ＤＮＡ，并与ＢＡ－ＥＬＩＳＡ检测胸水中抗ＰＰＤ抗体及细菌学检查相比较。结果：三种方法的敏感性和特异性分别为５６．０％、１００％；６４．０％、８６．７％、１１．０％和１００％。若以ＰＣＲ、ＢＡ－ＥＬＩＳＡ任一阳性作为判断标准则敏感性明显提高至８９．０％。认为，ＰＣＲ与ＢＡ－ＥＬＩＳＡ诊断结核性胸膜炎具有较高的敏感性，但前者更具早期和特异性并对疾病的疗效和预后有判断意义。     

聚合酶链反应快速诊断结核性胸腔积液的研究

聚合酶链反应快速诊断结核性胸腔积液的研究赵瑞贞，袁淑平，李冀文对７８例胸水标本行聚合酶链反应（ＰＣＲ）结核菌检测，以探明其诊断价值。结核性胸液组共５４例，男３９例，女１５例，年龄６～７８岁。诊断符合下列之一标准：（１）胸液、痰涂片和（或）结核菌培养（...     

胸膜活检标本行基因扩增对结核性胸膜炎的诊断价值

目的 评价胸膜活检组织行聚合酶链反应（ＰＣＲ）对结核性胸膜炎的诊断价值。方法 ＰＣＲ检测６５例胸膜活检组织中结核分枝杆菌ＤＮＡ,并与胸水检测及胸膜活检组织病检对比。结果 胸膜活检组织ＰＣＲ阳性率８３．１％,胸水ＰＣＲ阳性率为６３．１％,胸膜活检组织病检阳性率为６０．６％。前者较后两者更敏感。结论 胸膜活性组织ＰＣＲ检测对结核性胸膜炎有较高的诊断价值。     

聚合酶链反应诊断结核性腹膜为的评价

评价聚合酶链反应方法对结核性腹膜炎的诊断价值。方法 用ＰＣＲ技术检测３０例结核腹水中结核分支村菌ＤＮＡ。并与腹水涂片抗酸染色及酶联免疫吸附试验检测腹水中抗ＰＰＤ抗体进行比较。结果 ＰＣＲ的阳性率为６０％，特异性９４．４％，ＥＬＩＳＡ法阳生率６３．３％，特异性７２．２％涂片镜检均为阴性。     

Polymerase chain reaction of pleural biopsy is a rapid and sensitive method for the diagnosis of tuberculous pleural effusion

BACKGROUND: Tuberculous pleural effusion occurs in 30% of patients with tuberculosis (TB). Rapid diagnosis of a tuberculous pleural effusion would greatly facilitate the management of many patients. Polymerase chain reaction (PCR) has been used to detect Mycobacterium tuberculosis in pleural fluid with highly variable sensitivity. OBJECTIVE: To improve our laboratory diagnosis of tuberculous pleural effusion. METHODS: We applied PCR to detect DNA specific for M tuberculosis in 33 of the studied pleural biopsy specimens using an IS986-based primer that was specific for mycobacterium complex, and compared it to the results of pleural fluid and biopsy cultures performed on either Lowenstein-Jensen (LJ) medium or BACTEC 12B liquid medium (Becton Dickinson Microbiology Systems; Cockeysville, MD), Ziehl-Neelsen (ZN) staining, and histopathology in 45 patients with pleural effusion. RESULTS: Of the 45 patients with pleural effusion who were studied, 26 patients received diagnoses of tuberculous pleural effusion that had been confirmed by either culture and or histopathology, 10 patients received diagnoses of exudative effusion due to causes other than TB, and 9 patients received diagnoses of transudative effusion. Histopathology of the pleural biopsy specimen had a sensitivity of 53.8%. The sensitivity of the ZN staining of pleural fluid and biopsy specimens was 0.0% and 3.8%, respectively. The sensitivity of the culture on both BACTEC 12B liquid medium and LJ medium was higher in pleural biopsy specimens (92.3%) than in pleural fluid specimens (15.4%; p > 0.001). The improvements of the BACTEC culture system improved and shortened the detection time of M tuberculosis in pleural biopsy specimens. PCR of pleural biopsy specimens had 90% sensitivity and 100% specificity. The positive predictive value and the negative predictive value for pleural biopsy specimen cultures were 100% and 90.5% vs 100% and 86.7% for pleural biopsy specimen PCRs. CONCLUSION: The overall accuracy of PCR of pleural biopsy was similar to the results of pleural biopsy culture, however, PCR of the pleural biopsy was much faster in reaching diagnosis. PCR of pleural biopsy is a useful method when used in combination with the BACTEC culture system and histopathologic examination of pleural biopsy to reach a rapid diagnosis of tuberculous pleural effusion.     

Evaluation of polymerase chain reaction for detection of Mycobacterium tuberculosis in pleural fluid

OBJECTIVES: Tuberculosis, a reemergent killer, is threatening to assume serious proportions all over the world, particularly in view of the AIDS pandemic. The detection of mycobacterial DNA by polymerase chain reaction (PCR) in clinical samples is a promising approach for the rapid diagnosis of tuberculous infections. The aims of this study were to evaluate PCR for detection of Mycobacterium tuberculosis in pleural fluids and to correlate the results with adenosine deaminase activity (ADA) estimation and acid-fast bacilli (AFB) screening. METHODS: The sensitivity and specificity of PCR in detection of mycobacterial DNA in 20 samples of tuberculous pleural effusion were evaluated using 40 samples of nontubercular pleural effusion as controls. The results were correlated with the ADA in all 60 pleural fluids. In addition, AFB detection by Ziehl-Neelsen staining on cytospin smears of all pleural fluids was also compared. RESULTS: Of the 20 samples of tuberculous pleural effusion, mycobacterium could be detected by AFB staining in 4 samples. Fourteen samples were PCR positive. None of the samples from the control group were AFB or PCR positive. The sensitivity of PCR, therefore, was 70.0% with specificity of 100% (positive predictive value, 100%; negative predictive value, 86.95%). The sensitivity of AFB screening was at best 20%. The mean of ADA values in tubercular pleural effusions was 63.21 U/L (SD, 33.01), and the mean in the control samples was 51.1 U/L (SD, 29.71). Taking a cut-off value of 50 U/L, both the sensitivity and specificity of ADA estimation in diagnosing tuberculosis were only 55%. CONCLUSION: PCR represents a rapid and sensitive method for the detection of mycobacterial DNA in tuberculous pleural effusions. AFB screening has low sensitivity, and ADA estimation has both low sensitivity and specificity. Therefore, when the clinical suspicion is high and smear result is negative, but the signs and symptoms of M tuberculosis are apparent, PCR is the method of choice for identifying the infection.     

胸水和血清ADA、TB-Ab-IgG联合检测对结核性胸膜炎的诊断探讨

目的 探讨胸水/血清腺苷脱氨酶(ADA)、结核抗体(TB-Ab-IgG)联合检测对结核性胸膜炎的诊断价值.方法 采用斑点金免疫渗滤试验(DIGFA)和酶连续监测法对234例胸腔积液进行胸水/血清ADA和TB-Ab-IgG检测结果进行分析.结果 结核性胸膜炎患者174例其胸水、血清中TB-Ab-IgG的阳性率分别为62.0%和70.1%,特异性分别为93.1%(56/60)和86.6%(52/60).ADA活性在结核性和癌性胸腔积液中分别为(59.58±29.85)U/L和(15.31±7.36)U/L(P<0.01).以P-ADA>40 U/L做为诊断结核的临界值,其敏感性为79.3%,特异性为86.4%;以P-ADA/S-ADA>1为临界值,其敏感性为97.7%,特异性为95.5%.结论 胸水和血清ADA、TB-Ab-IgG联合检测在结核性胸膜炎与非结核性胸膜炎上具有诊断与鉴别诊断价值.     

结核γ-干扰素酶联免疫斑点检测在结核性心包积液诊断中的应用

目的 探讨结核菌特异性γ-干扰素（interferon-gamma,IFN-γ）酶联免疫斑点（enzyme-linked immunosorbent spot,ELISPOT）检测对结核性心包积液的诊断价值.方法 采用结核菌特异性IFN-γ ELISPOT技术同时检测20例结核性心包积液患者（TP组）和14例非结核性心包积液患者（Non-TP组）外周血单核细胞（peripheral blood mononuclear cells,PBMC）及心包积液单核细胞（pleural effusion mononuclear cells,PEMC）中结核菌特异性IFN-γ水平.结果 TP组PBMC和PEMC结核菌特异性IFN-γ水平均显著高于Non-TP组,差异有统计学意义（P＜0.01,P＜0.01）.TP组心包积液中结核菌特异性IFN-y水平显著高于外周血IFN-γ水平,差异有统计学意义（P＜0.01,P＜0.01）.PBMC ELISPOT检测结核性心包积液的敏感性和特异性分别为80.0％和85.7％;而PEMC ELISPOT敏感性和特异性为90.0％和85.7％.结论 结核菌特异性IFN-γELISPOT技术对结核性心包积液诊断和鉴别诊断具有很好的辅助价值.     

血清及胸水结核抗体联合检测对结核性胸膜炎的诊断价值

目的探讨血清及胸水结核抗体联合检测对结核性胸膜炎的诊断价值。方法联合检测118例结核性胸膜炎患者的血清及胸水结核抗体,同时随机选择47例非结核性胸膜炎患者为对照组。观察联合检测的敏感性,并与PPD试验相比较。结果结核性胸膜炎患者血清及胸水结核抗体阳性率(79.7%,70.3%)明显高于结核性胸膜炎患者PPD阳性率(56.8%)(P<0.05);血清及胸水结核抗体联合检测的敏感性(89.8%)明显高于PPD试验的敏感性(56.8%)(P<0.05)。结论血清及胸水结核抗体联合检测对结核性胸膜炎的诊断具有较高价值。     

Rapid diagnosis of tuberculous pleural effusion using polymerase chain reaction

Between October 1998 and September 1999, 98 patients with symptomatic exudative lymphocytic pleural effusion were enrolled in our study to evaluate the diagnostic sensitivity of polymerase chain reaction (PCR) assay. The mean age was 53.3 years ranging from 18 to 78 years. There were 61 men and 37 women. Pleural fluid was sent for gram staining, AFB staining, aerobic culture, culture of Mycobacterium tuberculosis on LJ media, and cytology. Additional fluid was used for a PCR-assay of the 16 S-23 S rRNA gene spacer sequences and for a nested PCR of the 16 S rRNA gene as a blind control. In cases of free-flow pleural tapping, histopathological analysis was done on three pleural biopsies. Overall etiologies comprised malignancy 53.1%, tuberculosis 36.7%, lymphoma 2.0% and chronic nonspecific inflammation 8.2%. The sensitivity and specificity of AFB-staining were 6% and 79%, respectively; while cultures on LJ media were 17% and 100%, respectively. The sensitivity of the PCR-assay was 50% (95% CI: 40 to 60%) and the specificity was 61% (95 CI: 52 to 71%). When PCR was nested, the sensitivity was 72% (95% CI: 63 to 81%) and specificity was 53% (95% CI: 43 to 63%). Two thirds (26 of 36) of tuberculous pleural effusion cases underwent pleural biopsy, and 62% were diagnosed by histopathology. There were no complications from thoracocentesis or pleural biopsy in any of the patients. We concluded that PCR assay was more sensitive than AFB staining and mycobacteria culture for diagnosis tuberculous pleural effusion but its specificity was quite low.     

结核性与癌性胸腔积液的实验室检测比较研究

目的比较研究实验室检测腺苷脱氨酶(ADA)、乳酸脱氢酶(LDH)、癌胚抗原(CEA)、蛋白(TP)、葡萄糖(GLU)等多项指标对结核性与癌性胸腔积液的鉴别诊断价值。方法对151例明确诊断为结核性或癌性胸腔积液分别测定胸水ADA、LDH、CEA、TP、GLU和血清TP,并进行统计分析。结果结核性胸腔积液中ADA、LDH、TP含量都明显高于癌性胸腔积液,其中胸水ADA以28U/L作为诊断结核性胸水的临界值则其敏感性和特异性均极高,结核性胸水中GLU含量则低于癌性胸水,癌性胸水CEA的阳性率高达76.0%,而结核性胸水CEA均阴性。结论联合检测胸水ADA、LDH、CEA、TP和GLU可以作为结核性与癌性胸腔积液的诊断和鉴别诊断依据,其中ADA28U/L可以考虑作为结核性胸腔积液的单独诊断依据。     

免疫组织化学及PCR技术在淋巴结结核病理诊断中的应用价值

目的 探讨免疫组织化学(immunohistochemistry,IHC)及PCR技术在淋巴结结核病理学诊断中的应用价值。方法 收集首都医科大学附属北京胸科医院病理科保存的2012年1月至2013年7月之间的48例手术根治切除淋巴结结核患者(结核组)和21例非淋巴结结核患者(非结核组)的石蜡包埋组织标本,分别应用IHC染色、荧光定量PCR和抗酸染色法对标本进行检测,以临床最后诊断为金标准,比较各方法的检测效能。结果 IHC染色、荧光定量PCR及抗酸染色检测的敏感度分别为52.1%(25/48)、60.4%(29/48)及27.1%(13/48);IHC染色和荧光定量PCR检测敏感度均高于抗酸染色,差异均有统计学意义(χ ²值分别为 6.27、10.84,P值分别为0.012、0.001);而IHC染色与荧光定量PCR比较,敏感度差异无统计学意义(χ ²=0.68,P=0.411)。IHC染色、荧光定量PCR及抗酸染色法检测非结核组结果均为阴性,特异度均为100%(21/21)。IHC染色、荧光定量PCR及抗酸染色法的阴性预测值分别为47.7%(21/44)、52.5%(21/40)、37.5%(21/56);符合率分别为66.7%(46/69)、72.5%(50/69)、49.3%(34/69),IHC染色、荧光定量PCR均优于抗酸染色法。 结论 IHC染色、荧光定量PCR与抗酸染色法相比,可提高阳性检出率,在淋巴结结核的病理学诊断中具有良好应用价值。