Evidence of Mycobacterium tuberculosis Complex bacteraemia in intradermal skin test positive cattle detected using phage-RPA

Bovine tuberculosis is a zoonotic infectious disease caused by Mycobacterium bovis 19 that affects cattle and can cause tuberculosis in a range of wildlife animals. A 20 bacteriophage-based method combined with PCR (phage-PCR) has been recently used to 21 detect and identify viable pathogenic mycoba...

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Main Authors: Swift, Benjamin, Convery, Thomas, Rees, Catherine
Format: Article
Published: Taylor & Francis 2016
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Online Access:https://eprints.nottingham.ac.uk/33738/
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author Swift, Benjamin
Convery, Thomas
Rees, Catherine
author_facet Swift, Benjamin
Convery, Thomas
Rees, Catherine
author_sort Swift, Benjamin
building Nottingham Research Data Repository
collection Online Access
description Bovine tuberculosis is a zoonotic infectious disease caused by Mycobacterium bovis 19 that affects cattle and can cause tuberculosis in a range of wildlife animals. A 20 bacteriophage-based method combined with PCR (phage-PCR) has been recently used to 21 detect and identify viable pathogenic mycobacteria in the peripheral blood mononuclear 22 cells (PBMCs) of animals suffering from paratuberculosis. To adapt this method for the 23 detection of M. bovis in blood, a new isothermal DNA amplification protocol using 24 Recombinase Polymerase Amplification (RPA) was developed and was found to be able to 25 detect M. bovis BCG within 48 h, with a limit of detection of approximately 10 cells per 26 ml of blood for artificially inoculated blood samples. When blood samples (2 ml) from a 27 Single Comparative Cervical Intradermal Tuberculin (SCCIT)- negative beef herd were 28 tested, Mycobacterium tuberculosis complex (MTC) cells were not detected from any 29 (45) of the blood samples. However when blood samples from SCCIT-positive animals 30 were tested, viable MTC bacteria were detected in 66 % (27/41) of samples. Of these 41 31 animals sampled, 32 % (13) had visible lesions. In the visible lesion (VL) group, 85 % 32 (11/13) had detectable levels of MTC whereas only 57 % (16/28) of animals which had 33 no visible lesions (NVL) were found to have detectable mycobacteraemia. These results 34 indicated that this simple, rapid method can be applied for the study of M. bovis 35 infections. The frequency with which viable mycobacteria were detected in the 36 peripheral blood of SCCIT-positive animals changes the paradigm of this disease.
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spelling nottingham-337382020-05-04T17:51:27Z https://eprints.nottingham.ac.uk/33738/ Evidence of Mycobacterium tuberculosis Complex bacteraemia in intradermal skin test positive cattle detected using phage-RPA Swift, Benjamin Convery, Thomas Rees, Catherine Bovine tuberculosis is a zoonotic infectious disease caused by Mycobacterium bovis 19 that affects cattle and can cause tuberculosis in a range of wildlife animals. A 20 bacteriophage-based method combined with PCR (phage-PCR) has been recently used to 21 detect and identify viable pathogenic mycobacteria in the peripheral blood mononuclear 22 cells (PBMCs) of animals suffering from paratuberculosis. To adapt this method for the 23 detection of M. bovis in blood, a new isothermal DNA amplification protocol using 24 Recombinase Polymerase Amplification (RPA) was developed and was found to be able to 25 detect M. bovis BCG within 48 h, with a limit of detection of approximately 10 cells per 26 ml of blood for artificially inoculated blood samples. When blood samples (2 ml) from a 27 Single Comparative Cervical Intradermal Tuberculin (SCCIT)- negative beef herd were 28 tested, Mycobacterium tuberculosis complex (MTC) cells were not detected from any 29 (45) of the blood samples. However when blood samples from SCCIT-positive animals 30 were tested, viable MTC bacteria were detected in 66 % (27/41) of samples. Of these 41 31 animals sampled, 32 % (13) had visible lesions. In the visible lesion (VL) group, 85 % 32 (11/13) had detectable levels of MTC whereas only 57 % (16/28) of animals which had 33 no visible lesions (NVL) were found to have detectable mycobacteraemia. These results 34 indicated that this simple, rapid method can be applied for the study of M. bovis 35 infections. The frequency with which viable mycobacteria were detected in the 36 peripheral blood of SCCIT-positive animals changes the paradigm of this disease. Taylor & Francis 2016-05-19 Article PeerReviewed Swift, Benjamin, Convery, Thomas and Rees, Catherine (2016) Evidence of Mycobacterium tuberculosis Complex bacteraemia in intradermal skin test positive cattle detected using phage-RPA. Virulence . ISSN 2150-5608 Bacteraemia Bacteriophage Blood Bovine Tuberculosis Detection http://www.tandfonline.com/doi/full/10.1080/21505594.2016.1191729 doi:10.1080/21505594.2016.1191729 doi:10.1080/21505594.2016.1191729
spellingShingle Bacteraemia
Bacteriophage
Blood
Bovine Tuberculosis
Detection
Swift, Benjamin
Convery, Thomas
Rees, Catherine
Evidence of Mycobacterium tuberculosis Complex bacteraemia in intradermal skin test positive cattle detected using phage-RPA
title Evidence of Mycobacterium tuberculosis Complex bacteraemia in intradermal skin test positive cattle detected using phage-RPA
title_full Evidence of Mycobacterium tuberculosis Complex bacteraemia in intradermal skin test positive cattle detected using phage-RPA
title_fullStr Evidence of Mycobacterium tuberculosis Complex bacteraemia in intradermal skin test positive cattle detected using phage-RPA
title_full_unstemmed Evidence of Mycobacterium tuberculosis Complex bacteraemia in intradermal skin test positive cattle detected using phage-RPA
title_short Evidence of Mycobacterium tuberculosis Complex bacteraemia in intradermal skin test positive cattle detected using phage-RPA
title_sort evidence of mycobacterium tuberculosis complex bacteraemia in intradermal skin test positive cattle detected using phage-rpa
topic Bacteraemia
Bacteriophage
Blood
Bovine Tuberculosis
Detection
url https://eprints.nottingham.ac.uk/33738/
https://eprints.nottingham.ac.uk/33738/
https://eprints.nottingham.ac.uk/33738/