Use of PCR in routine diagnosis of treated and untreated pulmonary tuberculosis.

Yuen, Kwok‐Yung; Chan, K. S.; Chan, Charles; Ho, B. S. W.; Dai, LK; Chau, Pui Hing; Mh, Ng

doi:10.1136/jcp.46.4.318

Cited by 54 publications

(28 citation statements)

References 8 publications

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“…The 38-kDa antigen gene is duplicated in M. intracellulare but absent from M. avium (54). A PCR assay designed to amplify the M. tuberculosis 38-kDa antigen gene gave positive results in M. bovis and M. africanum species, as well as M. tuberculosis, but not in other atypical mycobacteria (75).…”

Section: Discussionmentioning

confidence: 99%

Gamma Interferon Responses Induced by a Panel of Recombinant and Purified Mycobacterial Antigens in Healthy, Non-Mycobacterium bovisBCG-Vaccinated Malawian Young Adults

Black

Weir

Chaguluka

et al. 2003

Clin Vaccine Immunol

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We have previously shown that young adults living in a rural area of northern Malawi showed greater gamma interferon (IFN-␥) responses to purified protein derivatives (PPD) prepared from environmental mycobacteria than to PPD from Mycobacterium tuberculosis. In order to define the mycobacterial species to which individuals living in a rural African population have been exposed and sensitized, we tested T-cell recognition of recombinant and purified antigens from M. tuberculosis (38 kDa, MPT64, and ESAT-6), M. bovis There is great variation in the protection that Mycobacterium bovis BCG vaccines can provide against tuberculosis in different locations (17). In two large randomized controlled studies of BCG vaccination in Malawi and the United Kingdom, we have shown that prior to the vaccination of previously unvaccinated young adults in Malawi, over 60% of individuals show gamma interferon (IFN-␥) responses to purified protein derivative (PPD) from M. tuberculosis and that BCG vaccination provides only a moderate increase in this T-cell response (8). When PPDs from a number of nontuberculous environmental (atypical) mycobacteria were used to stimulate whole-blood cultures from the same subjects, PPDs from members of the M. avium-M. intracellulare-M. scrofulaceum complex induced stronger IFN-␥ responses than PPD from M. tuberculosis (6). To further define the mycobacterial species to which these subjects had been exposed, we have now used a panel of recombinant and purified antigens, with known species distributions, to test IFN-␥ responses, as a measure of the type 1 T-cell response induced by mycobacterial antigens, in day 6 supernatants from diluted whole-blood cultures. We have also assessed the potential of these recombinant antigens for identifying infection with M. tuberculosis and M. leprae, by studying the association between the IFN-␥ response to these antigens and the skin test response to M. tuberculosis PPD.Most studies of T-cell recognition of individual mycobacterial antigens have used patients infected with pathogenic mycobacteria or the contacts of these patients. Our study is unusual in that non-BCG-vaccinated, human immunodeficiency virus-negative, healthy individuals living in a rural area of northern Malawi have been tested for their ability to make a type 1 cytokine response to a panel of seven recombinant mycobacterial antigens and one purified native mycobacterial antigen. These subjects were clinically well at the time of testing and were recruited in their villages and homes.

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Section: Discussionmentioning

confidence: 99%

Gamma Interferon Responses Induced by a Panel of Recombinant and Purified Mycobacterial Antigens in Healthy, Non-Mycobacterium bovisBCG-Vaccinated Malawian Young Adults

Black

Weir

Chaguluka

et al. 2003

Clin Vaccine Immunol

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“…Both live and dead bacteria may result in positive PCR assays (17,25). Despite negative bacterial cultures, the presence of bacterial DNA in clinical specimens has been shown for prolonged periods in septic arthritis (6,44), pulmonary tuberculosis (14,49), and leptospirosis (3,27). The significance of the persistence of DNA for long periods after antibiotic treatment is uncertain (17,25).…”

Section: Discussionmentioning

confidence: 99%

“…In patients with septic arthritis, bacterial DNA can be found in synovial tissue for long periods after cultures become negative (44). The persistence of bacterial DNA despite effective antibiotic treatment has also been demonstrated in patients with pulmonary tuberculosis (14,49) and leptospirosis (3,27). DNA is sufficiently stable that it can be amplified by PCR for long periods after bacteria are no longer viable (7,25).…”

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confidence: 95%

PCR Detection of Bacteria on Cardiac Valves of Patients with Treated Bacterial Endocarditis

et al. 2005

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We used broad-range PCR amplification and sequencing to detect and identify bacterial DNA in 156 valves of patients treated for infective endocarditis (IE). Bacterial DNA was found more frequently in patients who underwent valve replacement while on antibiotic treatment for IE (60%) than in patients who had completed antibiotic treatment for IE (37%; P ‫؍‬ 0.02). We found specific bacterial DNA in valves removed from 11 of 30 patients who had completed antibiotic treatment for IE. Six had no histological evidence of IE. The presence of DNA was significantly correlated with the presence of histologic lesions (P ‫؍‬ 0.001) and with the presence of bacteria detected by Gram staining (P < 0.001). Bartonella and streptococci were detected for much longer after antibiotic treatment by PCR than other species (P ‫؍‬ 0.047 and 0.04, respectively), and coagulase-negative staphylococci were detected for much shorter periods (P ‫؍‬ 0.02). The finding that bacterial DNA was more likely to be detected in valves of patients with active IE than in patients who had completed antibiotic treatment for IE shows that bacterial DNA is cleared slowly. There was no significant correlation between the duration of antibiotic therapy and the presence of bacterial DNA in valves. Since the persistence of bacterial DNA in valves does not necessarily indicate the persistence of viable bacteria, the detection of bacterial DNA in valves from IE patients should be interpreted with caution, in particular in those patients with a past history of treated IE.

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“…A number of investigators have reported the usefulness of nucleic acid amplification methods to detect M. tuberculosis from clinical specimens (1,3,5,10,15,22,27,29,30,33,37,41). A PCR-based Roche Amplicor Mycobacterium system (Roche, Basel, Switzerland) for detecting M. tuberculosis, M. avium and M. intracellulare and an rRNA amplification-based Gen-Probe Amplified Mycobacterium Tuberculosis Direct Test system (GenProbe Inc., San Diego, Calif., U.S.A.) for detecting M. tuberculosis are now commercially available (1, 2, 4, 6-9, 11, 12, 18-20, 22-24, 29, 31, 34-36, 38-40).…”

mentioning

confidence: 99%

Diagnostic Value of the Amplicor PCR Assay for Initial Diagnosis and Assessment of Treatment Response for Pulmonary Tuberculosis

Iinuma

Ichiyama

Yamori

et al. 1998

Microbiology and Immunology

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We evaluated the Amplicor PCR assay as an initial diagnostic tool on the basis of clinical diagnosis, and assessed this assay as a follow-up test for patients with pulmonary tuberculosis during chemotherapy. Of the 208 specimens from 155 patients who were bacteriologically and/or clinically diagnosed with active tuberculosis before chemotherapy, 144 were Amplicor PCR-positive (sensitivity, 69.2%), which was equal to the results of culturing. Among 89 specimens which showed positive results by smear and culturing, the Amplicor PCR assay detected 87 (97.8%), whereas among 55 specimens which showed smearnegative but culture-positive results, the Amplicor PCR assay detected 46 (83.6 %)(P= 0.003). No false positive results were found in the two systems (specificity, 100%, 120/120). The Amplicor PCR assay was also evaluated as a follow-up test using 926 specimens from 207 patients receiving active tuberculosis chemotherapy. Among 433 specimens which showed Amplicor-PCR positive, 222 (51.3%) were culture-negative. On the other hand, among 233 culture-positive specimens, only 12 (5.2%) were Amplicor PCR-negative. Therefore, this assay is useful for the rapid diagnosis of tuberculosis. The duration of Amplicor PCR-positive after culture-negative conversion was significantly associated with the presence of cavitary lesion, smearpositive specimens before treatment, and smear-positive specimens with negative cultures during chemotherapy.

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Use of PCR in routine diagnosis of treated and untreated pulmonary tuberculosis.

Cited by 54 publications

References 8 publications

Gamma Interferon Responses Induced by a Panel of Recombinant and Purified Mycobacterial Antigens in Healthy, Non-Mycobacterium bovisBCG-Vaccinated Malawian Young Adults

Gamma Interferon Responses Induced by a Panel of Recombinant and Purified Mycobacterial Antigens in Healthy, Non-Mycobacterium bovisBCG-Vaccinated Malawian Young Adults

PCR Detection of Bacteria on Cardiac Valves of Patients with Treated Bacterial Endocarditis

Diagnostic Value of the Amplicor PCR Assay for Initial Diagnosis and Assessment of Treatment Response for Pulmonary Tuberculosis

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