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首页> 外文期刊>Journal of Clinical Microbiology >Detection of Mycobacterium tuberculosis in clinical samples by using polymerase chain reaction and a nonradioactive detection system.
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Detection of Mycobacterium tuberculosis in clinical samples by using polymerase chain reaction and a nonradioactive detection system.

机译：使用聚合酶链反应和非放射性检测系统检测临床样本中的结核分枝杆菌。

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A test based on the polymerase chain reaction (PCR) was developed for the detection of the Mycobacterium tuberculosis complex in clinical samples. In this test, a 245-bp sequence of the insertion element IS986 was amplified and detected by agarose gel electrophoresis in the presence of ethidium bromide and by Southern blot and dot blot hybridization by using a 188-bp digoxigenin-labeled probe. We tested clinical specimens from 227 patients suspected of having tuberculosis. These included 102 cerebrospinal fluid, 48 sputum, 18 pleural fluid, 5 bronchoalveolar lavage, 18 blood, 7 pus, 8 bone marrow, and 6 urine samples and 15 tissue biopsy specimens. We also tested sputum samples from 75 patients with diseases other than tuberculosis. Sputum samples were first decontaminated, and all samples were treated with proteinase K-detergent solution to extract the DNA. Part of each sample was spiked with M. tuberculosis to provide a semiquantitative assay and to control for the loss of mycobacteria or interference with the PCR which may cause false-negative results. One femtogram of M. tuberculosis DNA could be detected. PCR was positive for all 32 culture-positive (for M. tuberculosis) and Ziehl-Neelsen staining (ZN)-positive samples, 10 of 12 culture-positive and ZN-negative samples, and all 4 culture-negative and ZN-positive samples. PCR detected M. tuberculosis complex bacteria in 35 of 178 culture- and ZN-negative samples. Clinical data supported the diagnosis of tuberculosis in the majority of the 35 patients from whom those samples were obtained.
机译：开发了一种基于聚合酶链反应（PCR）的测试，用于检测临床样本中的结核分枝杆菌复合物。在该测试中，插入元件IS986的245-bp序列被扩增并在存在溴化乙锭的情况下通过琼脂糖凝胶电泳进行检测，并通过使用188-bp地高辛配基标记的探针通过Southern杂交和斑点杂交进行检测。我们测试了227名怀疑患有肺结核的患者的临床标本。其中包括102例脑脊液，48例痰，18例胸膜液，5例支气管肺泡灌洗液，18例血液，7例脓液，8例骨髓，6份尿液样本和15份组织活检标本。我们还测试了75例非结核病患者的痰液样本。首先对痰液样本进行消毒，然后用蛋白酶K洗涤剂溶液处理所有样本以提取DNA。将每个样品的一部分掺入结核分枝杆菌，以提供半定量测定，并控制分枝杆菌的丢失或对PCR的干扰（可能会导致假阴性结果）。可以检测到一飞克结核分枝杆菌DNA。所有32个培养阳性（结核分枝杆菌）和Ziehl-Neelsen染色（ZN）阳性样品，12个培养阳性和ZN阴性样品中的10个以及所有4个培养阴性和ZN阳性样品中的PCR均为阳性。 PCR检测了178个培养物和ZN阴性样品中的35个中的结核分枝杆菌复杂细菌。临床数据支持了从这些样本中获取的35例患者中的大多数的结核病诊断。

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