Development of a loop-mediated isothermal amplification (LAMP) method for specific detection of Mycobacterium bovis.
Bovine tuberculosis (TB) caused by Mycobacterium bovis is a significant health threat to cattle and a zoonotic threat for humans in many developing countries. Rapid and accurate detection of M. bovis is fundamental for controlling the disease in animals and humans, and for the proper treatment of pa...
| Main Authors: | , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2021-01-01
|
| Series: | PLoS Neglected Tropical Diseases |
| Online Access: | https://doi.org/10.1371/journal.pntd.0008996 |
| _version_ | 1818898865707810816 |
|---|---|
| author | Thoko Flav Kapalamula Jeewan Thapa Mwangala Lonah Akapelwa Kyoko Hayashida Stephen V Gordon Bernard Mudenda Hang' Ombe Musso Munyeme Eddie Samuneti Solo Precious Bwalya Mirriam Ethel Nyenje Aki Tamaru Yasuhiko Suzuki Chie Nakajima |
| author_facet | Thoko Flav Kapalamula Jeewan Thapa Mwangala Lonah Akapelwa Kyoko Hayashida Stephen V Gordon Bernard Mudenda Hang' Ombe Musso Munyeme Eddie Samuneti Solo Precious Bwalya Mirriam Ethel Nyenje Aki Tamaru Yasuhiko Suzuki Chie Nakajima |
| author_sort | Thoko Flav Kapalamula |
| collection | DOAJ |
| description | Bovine tuberculosis (TB) caused by Mycobacterium bovis is a significant health threat to cattle and a zoonotic threat for humans in many developing countries. Rapid and accurate detection of M. bovis is fundamental for controlling the disease in animals and humans, and for the proper treatment of patients as one of the first-line anti-TB drug, pyrazinamide, is ineffective against M. bovis. Currently, there are no rapid, simplified and low-cost diagnostic methods that can be easily integrated for use in many developing countries. Here, we report the development of a loop-mediated isothermal amplification (LAMP) assay for specific identification of M. bovis by targeting the region of difference 4 (RD4), a 12.7 kb genomic region that is deleted solely in M. bovis. The assay's specificity was evaluated using 139 isolates comprising 65 M. bovis isolates, 40 M. tuberculosis isolates, seven M. tuberculosis complex reference strains, 22 non-tuberculous mycobacteria and five other bacteria. The established LAMP detected only M. bovis isolates as positive and no false positives were observed using the other mycobacteria and non-mycobacteria tested. Our LAMP assay detected as low as 10 copies of M. bovis genomic DNA within 40 minutes. The procedure of LAMP is simple with an incubation at a constant temperature. Results are observed with the naked eye by a color change, and there is no need for expensive equipment. The established LAMP can be used for the detection of M. bovis infections in cattle and humans in resource-limited areas. |
| first_indexed | 2024-12-19T19:38:52Z |
| format | Article |
| id | doaj.art-d459809092c04cc2beea00cd7e774e42 |
| institution | Directory Open Access Journal |
| issn | 1935-2727 1935-2735 |
| language | English |
| last_indexed | 2024-12-19T19:38:52Z |
| publishDate | 2021-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Neglected Tropical Diseases |
| spelling | doaj.art-d459809092c04cc2beea00cd7e774e422022-12-21T20:08:18ZengPublic Library of Science (PLoS)PLoS Neglected Tropical Diseases1935-27271935-27352021-01-01151e000899610.1371/journal.pntd.0008996Development of a loop-mediated isothermal amplification (LAMP) method for specific detection of Mycobacterium bovis.Thoko Flav KapalamulaJeewan ThapaMwangala Lonah AkapelwaKyoko HayashidaStephen V GordonBernard Mudenda Hang' OmbeMusso MunyemeEddie Samuneti SoloPrecious BwalyaMirriam Ethel NyenjeAki TamaruYasuhiko SuzukiChie NakajimaBovine tuberculosis (TB) caused by Mycobacterium bovis is a significant health threat to cattle and a zoonotic threat for humans in many developing countries. Rapid and accurate detection of M. bovis is fundamental for controlling the disease in animals and humans, and for the proper treatment of patients as one of the first-line anti-TB drug, pyrazinamide, is ineffective against M. bovis. Currently, there are no rapid, simplified and low-cost diagnostic methods that can be easily integrated for use in many developing countries. Here, we report the development of a loop-mediated isothermal amplification (LAMP) assay for specific identification of M. bovis by targeting the region of difference 4 (RD4), a 12.7 kb genomic region that is deleted solely in M. bovis. The assay's specificity was evaluated using 139 isolates comprising 65 M. bovis isolates, 40 M. tuberculosis isolates, seven M. tuberculosis complex reference strains, 22 non-tuberculous mycobacteria and five other bacteria. The established LAMP detected only M. bovis isolates as positive and no false positives were observed using the other mycobacteria and non-mycobacteria tested. Our LAMP assay detected as low as 10 copies of M. bovis genomic DNA within 40 minutes. The procedure of LAMP is simple with an incubation at a constant temperature. Results are observed with the naked eye by a color change, and there is no need for expensive equipment. The established LAMP can be used for the detection of M. bovis infections in cattle and humans in resource-limited areas.https://doi.org/10.1371/journal.pntd.0008996 |
| spellingShingle | Thoko Flav Kapalamula Jeewan Thapa Mwangala Lonah Akapelwa Kyoko Hayashida Stephen V Gordon Bernard Mudenda Hang' Ombe Musso Munyeme Eddie Samuneti Solo Precious Bwalya Mirriam Ethel Nyenje Aki Tamaru Yasuhiko Suzuki Chie Nakajima Development of a loop-mediated isothermal amplification (LAMP) method for specific detection of Mycobacterium bovis. PLoS Neglected Tropical Diseases |
| title | Development of a loop-mediated isothermal amplification (LAMP) method for specific detection of Mycobacterium bovis. |
| title_full | Development of a loop-mediated isothermal amplification (LAMP) method for specific detection of Mycobacterium bovis. |
| title_fullStr | Development of a loop-mediated isothermal amplification (LAMP) method for specific detection of Mycobacterium bovis. |
| title_full_unstemmed | Development of a loop-mediated isothermal amplification (LAMP) method for specific detection of Mycobacterium bovis. |
| title_short | Development of a loop-mediated isothermal amplification (LAMP) method for specific detection of Mycobacterium bovis. |
| title_sort | development of a loop mediated isothermal amplification lamp method for specific detection of mycobacterium bovis |
| url | https://doi.org/10.1371/journal.pntd.0008996 |
| work_keys_str_mv | AT thokoflavkapalamula developmentofaloopmediatedisothermalamplificationlampmethodforspecificdetectionofmycobacteriumbovis AT jeewanthapa developmentofaloopmediatedisothermalamplificationlampmethodforspecificdetectionofmycobacteriumbovis AT mwangalalonahakapelwa developmentofaloopmediatedisothermalamplificationlampmethodforspecificdetectionofmycobacteriumbovis AT kyokohayashida developmentofaloopmediatedisothermalamplificationlampmethodforspecificdetectionofmycobacteriumbovis AT stephenvgordon developmentofaloopmediatedisothermalamplificationlampmethodforspecificdetectionofmycobacteriumbovis AT bernardmudendahangombe developmentofaloopmediatedisothermalamplificationlampmethodforspecificdetectionofmycobacteriumbovis AT mussomunyeme developmentofaloopmediatedisothermalamplificationlampmethodforspecificdetectionofmycobacteriumbovis AT eddiesamunetisolo developmentofaloopmediatedisothermalamplificationlampmethodforspecificdetectionofmycobacteriumbovis AT preciousbwalya developmentofaloopmediatedisothermalamplificationlampmethodforspecificdetectionofmycobacteriumbovis AT mirriamethelnyenje developmentofaloopmediatedisothermalamplificationlampmethodforspecificdetectionofmycobacteriumbovis AT akitamaru developmentofaloopmediatedisothermalamplificationlampmethodforspecificdetectionofmycobacteriumbovis AT yasuhikosuzuki developmentofaloopmediatedisothermalamplificationlampmethodforspecificdetectionofmycobacteriumbovis AT chienakajima developmentofaloopmediatedisothermalamplificationlampmethodforspecificdetectionofmycobacteriumbovis |