The utility of polymerase chain reaction (PCR) in the diagnosis of pulmonary tuberculosis.

Department of Microbiology, University Hospital and Medical School, Valencia, Spain.
Chest (Impact Factor: 7.13). 07/1995; 107(6):1631-5.
Source: PubMed

ABSTRACT A fragment of DNA of 123 bp belonging to insertion sequence IS6110, specific of Mycobacterium tuberculosis complex, was amplified by polymerase chain reaction (PCR) of respiratory samples, for the diagnosis of pulmonary tuberculosis. A total of 314 samples (286 sputum and 28 bronchoalveolar lavages) from 242 patients were evaluated by PCR, and the results were compared with the those obtained by acid-fast-stained smears, culture, and clinical diagnosis. Mycobacterium tuberculosis was detected by PCR in 102 of 105 patients with clinical diagnosis of pulmonary tuberculosis. All smear and culture-positive samples were PCR positive. The sensitivity of PCR, culture, and staining was 97%, 88%, and 65%, respectively, and the specificity was 100% in all cases. In ten patients with old residual lesions, but no active disease, M tuberculosis genome was detected by PCR. In our experience, PCR proved to be a useful method for the rapid diagnosis of pulmonary tuberculosis.

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    ABSTRACT: Background: It is difficult to diagnose Mycobacterium tuberculosis infection due to a lack of rapid, sensitive, and specific tests. Newer methods, which are easy and reliable, are required to diagnose TB at an early stage. Our aim is to evaluate the polymerase chain reaction (PCR) technique, using primers directed against the IS6110 gene, for the detection of M. tuberculosis in the sputum samples, and calculate the sensitivity and specificity of PCR. Patients and methods: A total of 248 sputum samples from patients suspected of mycobacterial diseases were studied. DNA was extracted by boiling method. IS6110 PCR method by a specific pair of primers designed to amplify 123bp and 245bp sequences of the insertion sequence, 6110, in the M. tuberculosis genome was used to analyze sputum samples. Results: Totally, 32 (12.9%) samples had positive culture. PCR yielded a sensitivity of 93.8% and specificity of 99.1% for the diagnosis of TB, when diagnosis was confirmed by culture. There were 2 out of 32(6.3%) PCR-positive cases among the patients with non-TB disease. Conclusion: We concluded that the performance of an IS6110 PCR assay is valuable in the rapid diagnosis of tuberculosis.
    Iranian Journal of Clinical Infectious Disease Iranian Journal of Clinical Infectious DiseasesIranian Journal of Clinical Infectious Diseases. 01/2009; 444:228-232228.
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    ABSTRACT: In developing countries the diagnosis of extrapulmonary tuberculosis (EPTB) is a major burning challenge. EPTB encounters many problems like pauci-bacillary nature, inadequate specimen volume. All the limitations reflect in the poor contribution of conventional bacteriological technique in the establishment of diagnosis of EPTB. Nucleic acid amplification methods are rapid and sensitive has modified strategies for the detection of mycobacterial DNA. A fragment of DNA of 123 bp belonging to insertion sequence IS6110 based on specific gene of Mycobacterium tuberculosis complex was amplified by polymerase chain reaction (PCR) for the rapid diagnosis of EPTB. The present study was to comparative evaluation of IS6110 PCR via conventional methods in the rapid diagnosis of new and Previously treated cases of extra pulmonary tuberculosis. Four hundred fifty specimens were collected from suspected cases of EPTB were processed for Mycobacteria by Zeihl Neelson (ZN) staining and BACTEC culture for M. tuberculosis. All the specimens were also processed for IS6110 based PCR amplification with primers targeting 123 bp fragment of insertion element IS6110 of M. tuberculosis complex. We found significant difference was seen in sensitivities of different tests. Of these 450 specimens, 60 (13.4%) were positive for AFB by ZN staining, 202 (45%) for BACTEC culture and IS6110 PCR were positive for M. tuberculosis complex in 283 (63%) specimens (p< 0.05). However, there was no significant difference (p< 0.05) as far as specificity of different tests. We found that IS6110 PCR has higher sensitivity than smear microscopy and BACTEC culture in both cases of new cases as well as in previously treated cases. IS6110 PCR can be highly useful in diagnosis of new and treated cases of EPTB. It may facilitate therapeutic decisions for those with suspected of EPTB.
    Tuberkuloz ve toraks 09/2011; 59(3):213-20.
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    ABSTRACT: The polymerase chain reaction (PCR) has recently been investigated for detecting Mycobacterium tuberculosis in clinical specimens and appears to have significant potential as a rapid, sensitive, and specific assay for diagnosing infections caused by this organism. One of the remaining issues regarding the use of this technique is the extraction of DNA from clinical material prior to performing the PCR. It requires the use of DNA extraction method that can efficiently lyse mycobacterial cells and recover small amounts of DNA. The method of heating the sample in a boiling water bath to break down the bacterial cell wall and to release the DNA was compared with that of enzymatic lysis of bacteria and then phenol-chloroform extraction of DNA (Modified chemical method). Modified chemical method was better than just boiling the sample to remove DNA for amplification with PCR. In this study 140 sputum samples were collected from suspected prisoners at Dhaka Central Jail. All of those samples were prepared by two DNA extraction methods and examined by PCR, and the results were compared with the results of acid-fast stained smears and cultures.
    Edited by Imatvievici, 06/2012; LAP Lambert Academic Publisher., ISBN: 978-3-659-16144-5


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