Novel targeting of the lepB gene using PCR with confronting two-pair primers for simultaneous detection of Mycobacterium tuberculosis complex and Mycobacterium bovis

© 2016 The Authors. Tuberculosis (TB), caused by members of the Mycobacterium tuberculosis complex (MTC), is the leading cause of infectious disease-related mortality worldwide. The standard method for TB diagnosis usually requires long periods of mycobacteria cultivation, leading to delayed diagnos...

Full description

Saved in:
Bibliographic Details
Main Authors: Chayada Sitthidet Tharinjaroen, Sorasak Intorasoot, Usanee Anukool, Ponrut Phunpae, Bordin Butr-Indr, Santhasiri Orrapin, Sirikwan Sangboonruang, Surachet Arunothong, Boonchai Chaiyasirinroj, Naowarat Kunyanone, Watchara Kasinrerk, Khajornsak Tragoolpua
Format: Journal
Published: 2018
Subjects:
Online Access:https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=84957564222&origin=inward
http://cmuir.cmu.ac.th/jspui/handle/6653943832/55911
Tags: Add Tag
No Tags, Be the first to tag this record!
Institution: Chiang Mai University
id th-cmuir.6653943832-55911
record_format dspace
spelling th-cmuir.6653943832-559112018-09-05T03:11:22Z Novel targeting of the lepB gene using PCR with confronting two-pair primers for simultaneous detection of Mycobacterium tuberculosis complex and Mycobacterium bovis Chayada Sitthidet Tharinjaroen Sorasak Intorasoot Usanee Anukool Ponrut Phunpae Bordin Butr-Indr Santhasiri Orrapin Sirikwan Sangboonruang Surachet Arunothong Boonchai Chaiyasirinroj Naowarat Kunyanone Watchara Kasinrerk Khajornsak Tragoolpua Immunology and Microbiology Medicine © 2016 The Authors. Tuberculosis (TB), caused by members of the Mycobacterium tuberculosis complex (MTC), is the leading cause of infectious disease-related mortality worldwide. The standard method for TB diagnosis usually requires long periods of mycobacteria cultivation, leading to delayed diagnosis, inefficient treatment and widespread occurrence of the disease. Therefore, a rapid method for the detection and differentiation of MTC from other mycobacteria is essential for disease diagnosis. Here, we describe the potential of using the type I signal peptidase (lepB) gene as a novel target for TB diagnosis, based on confronting two-pair primers PCR (PCRCTPP) that can detect MTC and simultaneously differentiate M. bovis. The limit of detection of the developed technique was equivalent to 12–120 bacilli. PCR-CTPP was highly specific to only MTC and M. bovis, and no cross-reaction was detected in 27 DNA of the non-tuberculous mycobacterial and bacterial strains tested. Thirty-nine blinded clinical isolates and 72 sputum samples were used to validate the PCR-CTPP in comparison with the standard mycobacterial culture method. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of PCR-CTPP were equal to 95, 100, 100 and 95%, respectively, when tested with clinical isolates. Furthermore, upon testing with the sputum samples, the sensitivity, specificity, PPV and NPV were observed to be 84, 76, 90 and 67%, respectively. Hence, this highly sensitive novel technique, which is rapid, easy to conduct and cost-effective, is a potential method for TB diagnosis and epidemiological studies, especially in resource-limited countries with a high TB burden. 2018-09-05T03:03:41Z 2018-09-05T03:03:41Z 2016-01-01 Journal 00222615 2-s2.0-84957564222 10.1099/jmm.0.000188 https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=84957564222&origin=inward http://cmuir.cmu.ac.th/jspui/handle/6653943832/55911
institution Chiang Mai University
building Chiang Mai University Library
country Thailand
collection CMU Intellectual Repository
topic Immunology and Microbiology
Medicine
spellingShingle Immunology and Microbiology
Medicine
Chayada Sitthidet Tharinjaroen
Sorasak Intorasoot
Usanee Anukool
Ponrut Phunpae
Bordin Butr-Indr
Santhasiri Orrapin
Sirikwan Sangboonruang
Surachet Arunothong
Boonchai Chaiyasirinroj
Naowarat Kunyanone
Watchara Kasinrerk
Khajornsak Tragoolpua
Novel targeting of the lepB gene using PCR with confronting two-pair primers for simultaneous detection of Mycobacterium tuberculosis complex and Mycobacterium bovis
description © 2016 The Authors. Tuberculosis (TB), caused by members of the Mycobacterium tuberculosis complex (MTC), is the leading cause of infectious disease-related mortality worldwide. The standard method for TB diagnosis usually requires long periods of mycobacteria cultivation, leading to delayed diagnosis, inefficient treatment and widespread occurrence of the disease. Therefore, a rapid method for the detection and differentiation of MTC from other mycobacteria is essential for disease diagnosis. Here, we describe the potential of using the type I signal peptidase (lepB) gene as a novel target for TB diagnosis, based on confronting two-pair primers PCR (PCRCTPP) that can detect MTC and simultaneously differentiate M. bovis. The limit of detection of the developed technique was equivalent to 12–120 bacilli. PCR-CTPP was highly specific to only MTC and M. bovis, and no cross-reaction was detected in 27 DNA of the non-tuberculous mycobacterial and bacterial strains tested. Thirty-nine blinded clinical isolates and 72 sputum samples were used to validate the PCR-CTPP in comparison with the standard mycobacterial culture method. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of PCR-CTPP were equal to 95, 100, 100 and 95%, respectively, when tested with clinical isolates. Furthermore, upon testing with the sputum samples, the sensitivity, specificity, PPV and NPV were observed to be 84, 76, 90 and 67%, respectively. Hence, this highly sensitive novel technique, which is rapid, easy to conduct and cost-effective, is a potential method for TB diagnosis and epidemiological studies, especially in resource-limited countries with a high TB burden.
format Journal
author Chayada Sitthidet Tharinjaroen
Sorasak Intorasoot
Usanee Anukool
Ponrut Phunpae
Bordin Butr-Indr
Santhasiri Orrapin
Sirikwan Sangboonruang
Surachet Arunothong
Boonchai Chaiyasirinroj
Naowarat Kunyanone
Watchara Kasinrerk
Khajornsak Tragoolpua
author_facet Chayada Sitthidet Tharinjaroen
Sorasak Intorasoot
Usanee Anukool
Ponrut Phunpae
Bordin Butr-Indr
Santhasiri Orrapin
Sirikwan Sangboonruang
Surachet Arunothong
Boonchai Chaiyasirinroj
Naowarat Kunyanone
Watchara Kasinrerk
Khajornsak Tragoolpua
author_sort Chayada Sitthidet Tharinjaroen
title Novel targeting of the lepB gene using PCR with confronting two-pair primers for simultaneous detection of Mycobacterium tuberculosis complex and Mycobacterium bovis
title_short Novel targeting of the lepB gene using PCR with confronting two-pair primers for simultaneous detection of Mycobacterium tuberculosis complex and Mycobacterium bovis
title_full Novel targeting of the lepB gene using PCR with confronting two-pair primers for simultaneous detection of Mycobacterium tuberculosis complex and Mycobacterium bovis
title_fullStr Novel targeting of the lepB gene using PCR with confronting two-pair primers for simultaneous detection of Mycobacterium tuberculosis complex and Mycobacterium bovis
title_full_unstemmed Novel targeting of the lepB gene using PCR with confronting two-pair primers for simultaneous detection of Mycobacterium tuberculosis complex and Mycobacterium bovis
title_sort novel targeting of the lepb gene using pcr with confronting two-pair primers for simultaneous detection of mycobacterium tuberculosis complex and mycobacterium bovis
publishDate 2018
url https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=84957564222&origin=inward
http://cmuir.cmu.ac.th/jspui/handle/6653943832/55911
_version_ 1681424593743511552