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TaqMan聚合酶链反应技术检测结核分支杆菌DNA及其临?… 总被引:22,自引:1,他引:22
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 相似文献
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结核分支杆菌DNA的单管巢式聚合酶链反应检测 总被引:8,自引:1,他引:8
目的探讨单管巢式聚合酶链反应(SNPCR)检测石蜡包埋组织结核分支杆菌DNA的特异性和敏感性。方法应用普通PCR(GPCR)、双管巢式PCR(DNPCR)和SNPCR对结核分支杆菌BCG和30例结核性淋巴结炎石蜡包埋组织进行结核分支杆菌复合群IS6110特异插入序列片段DNA检测。结果DNPCR和SNPCR检测BCGDNA均于15fg以上呈现阳性结果,其敏感性明显优于GPCR(480fg)。GPCR、DNPCR和SNPCR检测30例结核性淋巴结炎阳性率分别为43%、100%和100%,抗酸染色阳性率(10%)与3种PCR法相比差异有非常显著意义(均P<0.01)。GPCR阳性率与DNPCR和SNPCR相比差异亦具非常显著意义(均P<0.01)。SNPCR阳性率与DNPCR相同。结论巢式PCR检测淋巴结石蜡包埋组织结核分支杆菌的敏感性显著高于GPCR,其中SNPCR具有与DNPCR相同的特异性和敏感性,并具有更大的实用价值。 相似文献
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本文应用聚合酶链反应(PCR)技术检测了21例慢性肝炎患者福尔马林固定、石蜡包埋肝穿活检组织中的HBV DNA,并与其中的16例新鲜肝组织HBV DNA检出率进行了对照研究。结果显示:福尔马林固定、石蜡包埋肝穿刺活检组织HBV DNA检出率为71.43%(15/21),新鲜肝穿组织中HBV DNA检出率为87.50%(14/16),两者比较P>0.05。本文结果表明:(PCR)技术检测福尔马林固定、石蜡包埋肝穿刺活捡组织中的HBV DNA,具有材料来源广,易于保存等优点,适用于回顾性研究。 相似文献
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AmpliSensor—聚合酶链反应定量检测肺结核患者外周血结 … 总被引:14,自引:3,他引:11
目的 探讨AmpliSensor-聚合酶链反应定量检测外周血中结核分支杆菌DNA在肺结核的应用价值。方法 采用QlAamp和AcuPure法提取,制备全血中模板TB-DNA,应用AmpliSensor-PCR定量检测,并与IS6110-单管巢式聚合酶链反应(SN-PCR)作比较。结果200例肺结核患者的血液标本中,两种方法测得结核分支杆菌DNA的阳性率分别为60.5%、63.5%。85例非结核肺病 相似文献
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聚合酶链反应检测肺结核患者外周血单个核细胞中结核分支杆菌DNA的临床价值 总被引:9,自引:0,他引:9
应用聚合酶链反应(PCR)技术配对检测92例肺结核患者外周血单个核细胞内和痰标本中结核分支杆菌DNA,同时检测30例非结核患者外周血。结果显示:外周血和痰标本PCR阳性率分别为717%和554%,前者明显高于后者(P<005);各型肺结核外周血和痰标本PCR阳性率不尽相同;30例非结核患者外周血PCR阳性3例(10%)。认为PCR检测肺结核患者外周血单个核细胞内结核分支杆菌DNA是一种快速、敏感的方法,可用于肺结核早期诊断与鉴别诊断 相似文献
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聚合酶链反应检测肺结核患者外周血单个核细胞中结核分 … 总被引:8,自引:1,他引:8
探讨外周血单个核细胞中结核分支杆菌DNA检测在肺结核诊断中的价值。方法采用改良Triton-x-100法分离,制备单个核细胞中模板,DNA,聚合酶链反应扩增结核分支杆菌240bp基因片段,同时分析了影响PCR结果的有关因素。结果89例肺结核患者的血标本,84例肺结核患者的痰标本中,结核分支杆菌DNA阳性率分别为73%和57%;84例肺结核患者外周血,痰标本配对检测总阳性率可达87%。 相似文献
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DNA扩增抑制试验在聚合酶链反应检测结核分支杆菌中的作用张显忠郭爱军曹洪林杨明峰我们建立了DNA扩增抑制试验,检测系统中抑制因素所造成聚合酶链反应(PCR)试验的假阴性,以使PCR技术更加完善。材料与方法:标本来源:病例系本院住院肺结核病患者,根据临... 相似文献
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聚合酶链反应检测乙型肝炎患者心肌组织中HBV DNA 总被引:4,自引:0,他引:4
应用巢式聚合酶链反应(PCR)技术检测了18例乙型肝炎(乙肝)患者石蜡包埋心、肝组织中的HBV DNA,并与其免疫组化及原位杂交结果作了比较。二组引物检测的结果表明:在肝组织中均有HBV感染,其中5例有HBV复制。心肌组织中HBV DNA的检出率为55.6%,不存在HBV的复制。心脏病理检查可见一些非特异性改变,如脂变、水肿和纤维素样坏死等。病理变化与心电图、临床症状、体征及PCR检测结果无相关性 相似文献
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应用多重聚合酶链反应法检测并鉴别结核分支杆菌与非结核分支杆菌DNA 总被引:6,自引:1,他引:6
OBJECTIVE: To improve the specificity and sensitivity of polymerase chain reaction (PCR) technique for detecting and identification of DNA of M. tuberculosis and M. nontuberculosis. METHOD: Three pairs of oligonucleotide primer were used in triplex-PCR. A 383 bp DNA fragment encoding part of the 65,000 mycobacterial surface antigen, a 123 bp fragment corresponding to a specific M. tuberculosis complex sequence which was the insertion sequence 6110 (IS 6110) and a 268 bp fragment for human beta-globin were amplified by triplex-PCR respectively. RESULT: The sensitivity of the triplex-PCR-electrophoresis for the DNA of mycobacterium was 0.6 pg. The specific bands of 383 bp and 123 bp among the amplified DNA from M. hominis, M. bovis, BCG and M. simiae were present in the agarose gel. By contrast, only a band of 383 bp was found among the M. nontuberculosis which contained M. avium, M. chelonae, M. scrofulaceum, M. xenopi, M. kansasii, M. intracellulare and M. smegmatis. Compared with the standard strains, there was an additional 268 bp band in simulated clinical samples infected by Mycobacterium. The above 3 specific bands were found neither in other 15 bacterial species tested nor in Mycoplasma pneumoniae. 182 clinical samples were examined by culture, smear and triplex-PCR. 72 nontuberculous clinical samples were all negative. In 110 tuberculosis clinical samples, the positive rates were 2.7%, 13.6% and 32.7%, respectively. CONCLUSION: The triplex-PCR possesses a high specificity and sensitivity. This method could detect and identify the DNA of M. tuberculosis and M. nontuberculosis except M. simiae. It is a valuable tool for early diagnosis and differentiation for infection of M. tuberculosis and M. nontuberculosis. 相似文献
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实时定量聚合酶链反应技术在鉴别结节病与增殖性结核病中的应用 总被引:2,自引:0,他引:2
目的利用实时定量聚合酶链反应(PCR)方法检测结节病和结核病病理组织中结核分枝杆菌DNA,以探讨结核分枝杆菌在结节病发病中的作用及本方法在结节病与增殖性结核病鉴别中的应用价值。方法以上海市肺科医院1998年1月至2003年12月住院患者76例为研究对象,其中结核病组30例、结节病组31例和正常对照组(其他疾病)15例,利用定量PCR检测用石蜡包埋的淋巴结或肺组织中结核分枝杆菌DNA,并以胎鼠肺组织作为阴性对照。结果结核病组结核分枝杆菌DNA检出的阳性率(30/30)明显高于结节病组(6/31)和正常对照组(2/15),结节病组与正常对照组结核分枝杆菌DNA检出的阳性率无明显差别。结核病组结核分枝杆菌DNA检出的绝对和相对拷贝数明显高于结节病组和正常对照组,结节病组与正常对照组无明显差别,而胎鼠肺组织中未检出结核分枝杆菌DNA。结论本研究结果不支持结核分枝杆菌感染与结节病的相关性。实时定量PCR法检测石蜡包埋病理组织中结核分枝杆菌DNA可作为鉴别增殖性结核和结节病的方法之一。 相似文献
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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. 相似文献
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聚合酶链反应对菌阳肺结核治疗的监测 总被引:2,自引:2,他引:2
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。 相似文献
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Adriana Ant?nia da Cruz Furini Heloisa da Silveira Paro Pedro Jean Francisco Rodrigues Lilian Maria Lapa Montenegro Ricardo Luiz Dantas Machado Célia Franco Haiana Charifker Schindler Ida Maria Foschiani Dias Batista Andrea Regina Baptista Rossit 《Jornal brasileiro de pneumologia》2013,39(6):711-718
OBJECTIVE:
To compare the performance of nested polymerase chain reaction (NPCR) with that of cultures in the detection of the Mycobacterium tuberculosis complex in pulmonary and extrapulmonary specimens.METHODS:
We analyzed 20 and 78 pulmonary and extrapulmonary specimens, respectively, of 67 hospitalized patients suspected of having tuberculosis. An automated microbial system was used for the identification of Mycobacterium spp. cultures, and M. tuberculosis IS6110 was used as the target sequence in the NPCR. The kappa statistic was used in order to assess the level of agreement among the results.RESULTS:
Among the 67 patients, 6 and 5, respectively, were diagnosed with pulmonary and extrapulmonary tuberculosis, and the NPCR was positive in all of the cases. Among the 98 clinical specimens, smear microscopy, culture, and NPCR were positive in 6.00%, 8.16%, and 13.26%, respectively. Comparing the results of NPCR with those of cultures (the gold standard), we found that NPCR had a sensitivity and specificity of 100% and 83%, respectively, in pulmonary specimens, compared with 83% and 96%, respectively, in extrapulmonary specimens, with good concordance between the tests (kappa, 0.50 and 0.6867, respectively).CONCLUSIONS:
Although NPCR proved to be a very useful tool for the detection of M. tuberculosis complex, clinical, epidemiological, and other laboratory data should also be considered in the diagnosis and treatment of pulmonary and extrapulmonary tuberculosis. 相似文献16.
利用聚合酶链反应检测122例结核病患者标本中结核菌的研究 总被引:10,自引:0,他引:10
对122例门诊及住院结核病人的痰、脓汁等标本进行聚合酶链反应(PCR)和直接涂片荧光镜检,培养检查结核菌,结果表示:PCR阳性率为56.9%(58/102),培养阳性率为14.7%(15/102),直接涂片荧光镜检为23.5%(24/102)。说明PCR特异性好,敏感性强,与临床符合率较高。PCR比培养法大大节省时间,为结核病的诊断和鉴别赢得了时间,是一种可以推广的好方法。 相似文献
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16SrRNA基因PCR加反相杂交技术检测细菌DNA 总被引:13,自引:1,他引:12
目的探讨聚合酶链反应(PCR)加反相杂交技术在细菌DNA检测中的应用。方法以16SrRNA基因为靶序列,设计引物及寡核苷酸探针,采用PCR法加反相杂交检测标准菌株及临床标本细菌DNA。结果对20株不同标准菌株进行PCR扩增,均出现371bp长度的DNA片段,敏感性试验可检测出10-12g的细菌DNA,与人基因组及病毒无交叉反应;22份血培养阳性标本及4份脑脊液培养阳性标本均扩增出371bp长度DNA条带,反相杂交区分革兰阳性/阴性细菌与培养结果相符。结论16SrRNA基因PCR加反相杂交技术检测细菌DNA,具有敏感、快速、准确的特点,为细菌感染的临床诊断提供了科学的依据。 相似文献