A novel ready-to-use loop-mediated isothermal amplification (LAMP) method for detection of Burkholderia mallei and B. pseudomallei
Abstract Background Glanders and melioidosis are contagious zoonotic diseases caused by Burkholderia mallei and B. pseudomallei, respectively. Bacterial isolation and polymerase chain reaction (PCR) have been used to detect these bacteria in animals suspected of infection; however, both methods requ...
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BMC
2025-01-01
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| Series: | BMC Microbiology |
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| Online Access: | https://doi.org/10.1186/s12866-024-03737-z |
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| author | Mitsuru Nakase Jeewan Thapa Vanaabaatar Batbaatar Ochirbat Khurtsbaatar Batchuluun Enkhtuul Jugderkhorloo Unenbat Baasansuren Lkham Sachiho Fujita Ai Koshikawa Apichai Tuanyok Vannarat Saechan Hideaki Higashi Kyoko Hayashida Yasuhiko Suzuki Chie Nakajima Takashi Kimura |
| author_facet | Mitsuru Nakase Jeewan Thapa Vanaabaatar Batbaatar Ochirbat Khurtsbaatar Batchuluun Enkhtuul Jugderkhorloo Unenbat Baasansuren Lkham Sachiho Fujita Ai Koshikawa Apichai Tuanyok Vannarat Saechan Hideaki Higashi Kyoko Hayashida Yasuhiko Suzuki Chie Nakajima Takashi Kimura |
| author_sort | Mitsuru Nakase |
| collection | DOAJ |
| description | Abstract Background Glanders and melioidosis are contagious zoonotic diseases caused by Burkholderia mallei and B. pseudomallei, respectively. Bacterial isolation and polymerase chain reaction (PCR) have been used to detect these bacteria in animals suspected of infection; however, both methods require skilled experimental techniques and expensive equipment. These obstacles make it difficult to diagnose B. mallei and B. pseudomallei infections in areas where reagents and equipment are difficult to procure. To solve this problem, we developed an easy and ready-to-use dried-format diagnostic tool based on loop-mediated isothermal amplification (LAMP) method. Results The primer set targeting the internal transcribed spacer (ITS) region detected 10 genomic copies of B. mallei DNA and B. pseudomallei DNA using the conventional liquid LAMP method. This primer set did not detect any other Burkholderia species. Using this novel primer set, a dried-format in-house LAMP method with high sensitivity and specificity was developed. This method was used to test for the presence of B. mallei DNA in swabs collected from the nasal cavity and ulcerated skin of 19 B. mallei-infected horses and five uninfected horses and was compared with the real-time PCR method. These two tests showed 87.5% agreement for the positive samples and 100% agreement for the negative samples. This method detected all tested B. pseudomallei clinical isolates. Conclusions We established the first dry LAMP method for the detection of B. mallei and B. pseudomallei. This study provided a simple, rapid, cost-effective, and sensitive diagnostic tool for glanders and melioidosis. |
| format | Article |
| id | doaj-art-2e78229c94864a6eaee5b89868719480 |
| institution | Kabale University |
| issn | 1471-2180 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Microbiology |
| spelling | doaj-art-2e78229c94864a6eaee5b898687194802025-01-26T12:17:58ZengBMCBMC Microbiology1471-21802025-01-0125111310.1186/s12866-024-03737-zA novel ready-to-use loop-mediated isothermal amplification (LAMP) method for detection of Burkholderia mallei and B. pseudomalleiMitsuru Nakase0Jeewan Thapa1Vanaabaatar Batbaatar2Ochirbat Khurtsbaatar3Batchuluun Enkhtuul4Jugderkhorloo Unenbat5Baasansuren Lkham6Sachiho Fujita7Ai Koshikawa8Apichai Tuanyok9Vannarat Saechan10Hideaki Higashi11Kyoko Hayashida12Yasuhiko Suzuki13Chie Nakajima14Takashi Kimura15Laboratory of Comparative Pathology, Faculty of Veterinary Medicine, Hokkaido UniversityDivision of Bioresources, International Institute for Zoonosis Control, Hokkaido UniversityLaboratory of Infectious Disease and Immunology, Institute of Veterinary Medicine, Mongolian University of Life Science, Zaisan 17024, Khan-Uul District, Ulaanbaatar CityLaboratory of Infectious Disease and Immunology, Institute of Veterinary Medicine, Mongolian University of Life Science, Zaisan 17024, Khan-Uul District, Ulaanbaatar CityLaboratory of Infectious Disease and Immunology, Institute of Veterinary Medicine, Mongolian University of Life Science, Zaisan 17024, Khan-Uul District, Ulaanbaatar CityLaboratory of Infectious Disease and Immunology, Institute of Veterinary Medicine, Mongolian University of Life Science, Zaisan 17024, Khan-Uul District, Ulaanbaatar CityLaboratory of Infectious Disease and Immunology, Institute of Veterinary Medicine, Mongolian University of Life Science, Zaisan 17024, Khan-Uul District, Ulaanbaatar CityLaboratory of Comparative Pathology, Faculty of Veterinary Medicine, Hokkaido UniversityLaboratory of Comparative Pathology, Faculty of Veterinary Medicine, Hokkaido UniversityDepartment of Infectious Diseases and Immunology, College of Veterinary Medicine, University of FloridaFaculty of Veterinary Science, Prince of Songkla UniversityDivision of Infection and Immunity, International Institute for Zoonosis Control, Hokkaido UniversityDivision of Collaborations and Education, International Institute for Zoonosis Control, Hokkaido UniversityDivision of Bioresources, International Institute for Zoonosis Control, Hokkaido UniversityDivision of Bioresources, International Institute for Zoonosis Control, Hokkaido UniversityLaboratory of Comparative Pathology, Faculty of Veterinary Medicine, Hokkaido UniversityAbstract Background Glanders and melioidosis are contagious zoonotic diseases caused by Burkholderia mallei and B. pseudomallei, respectively. Bacterial isolation and polymerase chain reaction (PCR) have been used to detect these bacteria in animals suspected of infection; however, both methods require skilled experimental techniques and expensive equipment. These obstacles make it difficult to diagnose B. mallei and B. pseudomallei infections in areas where reagents and equipment are difficult to procure. To solve this problem, we developed an easy and ready-to-use dried-format diagnostic tool based on loop-mediated isothermal amplification (LAMP) method. Results The primer set targeting the internal transcribed spacer (ITS) region detected 10 genomic copies of B. mallei DNA and B. pseudomallei DNA using the conventional liquid LAMP method. This primer set did not detect any other Burkholderia species. Using this novel primer set, a dried-format in-house LAMP method with high sensitivity and specificity was developed. This method was used to test for the presence of B. mallei DNA in swabs collected from the nasal cavity and ulcerated skin of 19 B. mallei-infected horses and five uninfected horses and was compared with the real-time PCR method. These two tests showed 87.5% agreement for the positive samples and 100% agreement for the negative samples. This method detected all tested B. pseudomallei clinical isolates. Conclusions We established the first dry LAMP method for the detection of B. mallei and B. pseudomallei. This study provided a simple, rapid, cost-effective, and sensitive diagnostic tool for glanders and melioidosis.https://doi.org/10.1186/s12866-024-03737-zDry LAMPLoop-mediated isothermal amplificationBurkholderia malleiBurkholderia pseudomalleiGlandersMelioidosis |
| spellingShingle | Mitsuru Nakase Jeewan Thapa Vanaabaatar Batbaatar Ochirbat Khurtsbaatar Batchuluun Enkhtuul Jugderkhorloo Unenbat Baasansuren Lkham Sachiho Fujita Ai Koshikawa Apichai Tuanyok Vannarat Saechan Hideaki Higashi Kyoko Hayashida Yasuhiko Suzuki Chie Nakajima Takashi Kimura A novel ready-to-use loop-mediated isothermal amplification (LAMP) method for detection of Burkholderia mallei and B. pseudomallei BMC Microbiology Dry LAMP Loop-mediated isothermal amplification Burkholderia mallei Burkholderia pseudomallei Glanders Melioidosis |
| title | A novel ready-to-use loop-mediated isothermal amplification (LAMP) method for detection of Burkholderia mallei and B. pseudomallei |
| title_full | A novel ready-to-use loop-mediated isothermal amplification (LAMP) method for detection of Burkholderia mallei and B. pseudomallei |
| title_fullStr | A novel ready-to-use loop-mediated isothermal amplification (LAMP) method for detection of Burkholderia mallei and B. pseudomallei |
| title_full_unstemmed | A novel ready-to-use loop-mediated isothermal amplification (LAMP) method for detection of Burkholderia mallei and B. pseudomallei |
| title_short | A novel ready-to-use loop-mediated isothermal amplification (LAMP) method for detection of Burkholderia mallei and B. pseudomallei |
| title_sort | novel ready to use loop mediated isothermal amplification lamp method for detection of burkholderia mallei and b pseudomallei |
| topic | Dry LAMP Loop-mediated isothermal amplification Burkholderia mallei Burkholderia pseudomallei Glanders Melioidosis |
| url | https://doi.org/10.1186/s12866-024-03737-z |
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