author_facet Down, J A
O'Connell, M A
Dey, M S
Walters, A H
Howard, D R
Little, M C
Keating, W E
Zwadyk, P
Haaland, P D
McLaurin, D A
Cole, G
Down, J A
O'Connell, M A
Dey, M S
Walters, A H
Howard, D R
Little, M C
Keating, W E
Zwadyk, P
Haaland, P D
McLaurin, D A
Cole, G
author Down, J A
O'Connell, M A
Dey, M S
Walters, A H
Howard, D R
Little, M C
Keating, W E
Zwadyk, P
Haaland, P D
McLaurin, D A
Cole, G
spellingShingle Down, J A
O'Connell, M A
Dey, M S
Walters, A H
Howard, D R
Little, M C
Keating, W E
Zwadyk, P
Haaland, P D
McLaurin, D A
Cole, G
Journal of Clinical Microbiology
Detection of Mycobacterium tuberculosis in respiratory specimens by strand displacement amplification of DNA
Microbiology (medical)
author_sort down, j a
spelling Down, J A O'Connell, M A Dey, M S Walters, A H Howard, D R Little, M C Keating, W E Zwadyk, P Haaland, P D McLaurin, D A Cole, G 0095-1137 1098-660X American Society for Microbiology Microbiology (medical) http://dx.doi.org/10.1128/jcm.34.4.860-865.1996 <jats:p>A total of 294 clinical respiratory specimens, including 75 with culture-positive results, were tested for the presence of Mycobacterium tuberculosis by strand displacement amplification (SDA) of DNA. A region of the IS6110 insertion element and an internal control sequence were amplified and then detected by a chemiluminescence assay. Receiver operator-characteristic curves were used to evaluate three methods for declaring specimens positive for M. tuberculosis. By the preferred method, SDA chemiluminescence results were converted to theoretical numbers of M. tuberculosis organisms. A positive threshold (PT) value, above which 95% of the SDA results were judged to be M. tuberculosis positive (sensitivity = 95%), was found to be 2.4 M. tuberculosis organisms per SDA reaction. The analogous PT value for 95% sensitivity on smear-positive specimens was 3.6 M. tuberculosis organisms per reaction. The PT of 2.4 M. tuberculosis organisms per reaction detected 100% of culture-positive, smear-positive specimens (sensitivity = 100%), while 95% sensitivity was achieved with a PT of 15.5 M. tuberculosis organisms per reaction. Specificities, which were calculated with respect to culture- and smear-negative specimens, ranged from 96% at a PT of 15.5 M. tuberculosis organisms to 84% at a PT of 2.4 M. tuberculosis organisms per reaction. The M. tuberculosis-negative specimens were also segregated according to whether the patients received antituberculosis chemotherapy. SDA specificity ranged from 90% (PT = 2.4 M. tuberculosis organisms) to 98% (PT = 15.5 M. tuberculosis organisms) for the M. tuberculosis-negative specimens from patients who had not received chemotherapy. SDA specificity in the M. tuberculosis-negative specimens from patients who received chemotherapy was lower (85 to 94%). This study represents the first large-scale demonstration of M. tuberculosis detection in clinical sputum specimens by isothermal DNA amplification with SDA.</jats:p> Detection of Mycobacterium tuberculosis in respiratory specimens by strand displacement amplification of DNA Journal of Clinical Microbiology
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series Journal of Clinical Microbiology
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title Detection of Mycobacterium tuberculosis in respiratory specimens by strand displacement amplification of DNA
title_unstemmed Detection of Mycobacterium tuberculosis in respiratory specimens by strand displacement amplification of DNA
title_full Detection of Mycobacterium tuberculosis in respiratory specimens by strand displacement amplification of DNA
title_fullStr Detection of Mycobacterium tuberculosis in respiratory specimens by strand displacement amplification of DNA
title_full_unstemmed Detection of Mycobacterium tuberculosis in respiratory specimens by strand displacement amplification of DNA
title_short Detection of Mycobacterium tuberculosis in respiratory specimens by strand displacement amplification of DNA
title_sort detection of mycobacterium tuberculosis in respiratory specimens by strand displacement amplification of dna
topic Microbiology (medical)
url http://dx.doi.org/10.1128/jcm.34.4.860-865.1996
publishDate 1996
physical 860-865
description <jats:p>A total of 294 clinical respiratory specimens, including 75 with culture-positive results, were tested for the presence of Mycobacterium tuberculosis by strand displacement amplification (SDA) of DNA. A region of the IS6110 insertion element and an internal control sequence were amplified and then detected by a chemiluminescence assay. Receiver operator-characteristic curves were used to evaluate three methods for declaring specimens positive for M. tuberculosis. By the preferred method, SDA chemiluminescence results were converted to theoretical numbers of M. tuberculosis organisms. A positive threshold (PT) value, above which 95% of the SDA results were judged to be M. tuberculosis positive (sensitivity = 95%), was found to be 2.4 M. tuberculosis organisms per SDA reaction. The analogous PT value for 95% sensitivity on smear-positive specimens was 3.6 M. tuberculosis organisms per reaction. The PT of 2.4 M. tuberculosis organisms per reaction detected 100% of culture-positive, smear-positive specimens (sensitivity = 100%), while 95% sensitivity was achieved with a PT of 15.5 M. tuberculosis organisms per reaction. Specificities, which were calculated with respect to culture- and smear-negative specimens, ranged from 96% at a PT of 15.5 M. tuberculosis organisms to 84% at a PT of 2.4 M. tuberculosis organisms per reaction. The M. tuberculosis-negative specimens were also segregated according to whether the patients received antituberculosis chemotherapy. SDA specificity ranged from 90% (PT = 2.4 M. tuberculosis organisms) to 98% (PT = 15.5 M. tuberculosis organisms) for the M. tuberculosis-negative specimens from patients who had not received chemotherapy. SDA specificity in the M. tuberculosis-negative specimens from patients who received chemotherapy was lower (85 to 94%). This study represents the first large-scale demonstration of M. tuberculosis detection in clinical sputum specimens by isothermal DNA amplification with SDA.</jats:p>
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author Down, J A, O'Connell, M A, Dey, M S, Walters, A H, Howard, D R, Little, M C, Keating, W E, Zwadyk, P, Haaland, P D, McLaurin, D A, Cole, G
author_facet Down, J A, O'Connell, M A, Dey, M S, Walters, A H, Howard, D R, Little, M C, Keating, W E, Zwadyk, P, Haaland, P D, McLaurin, D A, Cole, G, Down, J A, O'Connell, M A, Dey, M S, Walters, A H, Howard, D R, Little, M C, Keating, W E, Zwadyk, P, Haaland, P D, McLaurin, D A, Cole, G
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description <jats:p>A total of 294 clinical respiratory specimens, including 75 with culture-positive results, were tested for the presence of Mycobacterium tuberculosis by strand displacement amplification (SDA) of DNA. A region of the IS6110 insertion element and an internal control sequence were amplified and then detected by a chemiluminescence assay. Receiver operator-characteristic curves were used to evaluate three methods for declaring specimens positive for M. tuberculosis. By the preferred method, SDA chemiluminescence results were converted to theoretical numbers of M. tuberculosis organisms. A positive threshold (PT) value, above which 95% of the SDA results were judged to be M. tuberculosis positive (sensitivity = 95%), was found to be 2.4 M. tuberculosis organisms per SDA reaction. The analogous PT value for 95% sensitivity on smear-positive specimens was 3.6 M. tuberculosis organisms per reaction. The PT of 2.4 M. tuberculosis organisms per reaction detected 100% of culture-positive, smear-positive specimens (sensitivity = 100%), while 95% sensitivity was achieved with a PT of 15.5 M. tuberculosis organisms per reaction. Specificities, which were calculated with respect to culture- and smear-negative specimens, ranged from 96% at a PT of 15.5 M. tuberculosis organisms to 84% at a PT of 2.4 M. tuberculosis organisms per reaction. The M. tuberculosis-negative specimens were also segregated according to whether the patients received antituberculosis chemotherapy. SDA specificity ranged from 90% (PT = 2.4 M. tuberculosis organisms) to 98% (PT = 15.5 M. tuberculosis organisms) for the M. tuberculosis-negative specimens from patients who had not received chemotherapy. SDA specificity in the M. tuberculosis-negative specimens from patients who received chemotherapy was lower (85 to 94%). This study represents the first large-scale demonstration of M. tuberculosis detection in clinical sputum specimens by isothermal DNA amplification with SDA.</jats:p>
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spelling Down, J A O'Connell, M A Dey, M S Walters, A H Howard, D R Little, M C Keating, W E Zwadyk, P Haaland, P D McLaurin, D A Cole, G 0095-1137 1098-660X American Society for Microbiology Microbiology (medical) http://dx.doi.org/10.1128/jcm.34.4.860-865.1996 <jats:p>A total of 294 clinical respiratory specimens, including 75 with culture-positive results, were tested for the presence of Mycobacterium tuberculosis by strand displacement amplification (SDA) of DNA. A region of the IS6110 insertion element and an internal control sequence were amplified and then detected by a chemiluminescence assay. Receiver operator-characteristic curves were used to evaluate three methods for declaring specimens positive for M. tuberculosis. By the preferred method, SDA chemiluminescence results were converted to theoretical numbers of M. tuberculosis organisms. A positive threshold (PT) value, above which 95% of the SDA results were judged to be M. tuberculosis positive (sensitivity = 95%), was found to be 2.4 M. tuberculosis organisms per SDA reaction. The analogous PT value for 95% sensitivity on smear-positive specimens was 3.6 M. tuberculosis organisms per reaction. The PT of 2.4 M. tuberculosis organisms per reaction detected 100% of culture-positive, smear-positive specimens (sensitivity = 100%), while 95% sensitivity was achieved with a PT of 15.5 M. tuberculosis organisms per reaction. Specificities, which were calculated with respect to culture- and smear-negative specimens, ranged from 96% at a PT of 15.5 M. tuberculosis organisms to 84% at a PT of 2.4 M. tuberculosis organisms per reaction. The M. tuberculosis-negative specimens were also segregated according to whether the patients received antituberculosis chemotherapy. SDA specificity ranged from 90% (PT = 2.4 M. tuberculosis organisms) to 98% (PT = 15.5 M. tuberculosis organisms) for the M. tuberculosis-negative specimens from patients who had not received chemotherapy. SDA specificity in the M. tuberculosis-negative specimens from patients who received chemotherapy was lower (85 to 94%). This study represents the first large-scale demonstration of M. tuberculosis detection in clinical sputum specimens by isothermal DNA amplification with SDA.</jats:p> Detection of Mycobacterium tuberculosis in respiratory specimens by strand displacement amplification of DNA Journal of Clinical Microbiology
spellingShingle Down, J A, O'Connell, M A, Dey, M S, Walters, A H, Howard, D R, Little, M C, Keating, W E, Zwadyk, P, Haaland, P D, McLaurin, D A, Cole, G, Journal of Clinical Microbiology, Detection of Mycobacterium tuberculosis in respiratory specimens by strand displacement amplification of DNA, Microbiology (medical)
title Detection of Mycobacterium tuberculosis in respiratory specimens by strand displacement amplification of DNA
title_full Detection of Mycobacterium tuberculosis in respiratory specimens by strand displacement amplification of DNA
title_fullStr Detection of Mycobacterium tuberculosis in respiratory specimens by strand displacement amplification of DNA
title_full_unstemmed Detection of Mycobacterium tuberculosis in respiratory specimens by strand displacement amplification of DNA
title_short Detection of Mycobacterium tuberculosis in respiratory specimens by strand displacement amplification of DNA
title_sort detection of mycobacterium tuberculosis in respiratory specimens by strand displacement amplification of dna
title_unstemmed Detection of Mycobacterium tuberculosis in respiratory specimens by strand displacement amplification of DNA
topic Microbiology (medical)
url http://dx.doi.org/10.1128/jcm.34.4.860-865.1996