A new primer construction technique that effectively increases amplification of rare mutant templates in samples
Abstract Background In personalized medicine, companion diagnostic tests provide additional information to help select a treatment option likely to be optimal for a patient. Although such tests include several techniques for detecting low levels of mutant genes in wild-type backgrounds with fairly h...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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BMC
2019-08-01
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| Series: | BMC Biotechnology |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12896-019-0555-1 |
| _version_ | 1818335008021020672 |
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| author | Jr-Kai Huang Ling Fan Tao-Yeuan Wang Pao-Shu Wu |
| author_facet | Jr-Kai Huang Ling Fan Tao-Yeuan Wang Pao-Shu Wu |
| author_sort | Jr-Kai Huang |
| collection | DOAJ |
| description | Abstract Background In personalized medicine, companion diagnostic tests provide additional information to help select a treatment option likely to be optimal for a patient. Although such tests include several techniques for detecting low levels of mutant genes in wild-type backgrounds with fairly high sensitivity, most tests are not specific, and may exhibit high false positive rates. In this study, we describe a new primer structure, named ‘stuntmer’, to selectively suppress amplification of wild-type templates, and promote amplification of mutant templates. Results A single stuntmer for a defined region of DNA can detect several kinds of mutations, including point mutations, deletions, and insertions. Stuntmer PCRs are also highly sensitive, being able to amplify mutant sequences that may make up as little as 0.1% of the DNA sample. Conclusion In conclusion, our technique, stuntmer PCR, can provide a simple, low-cost, highly sensitive, highly accurate, and highly specific platform for developing companion diagnostic tests. |
| first_indexed | 2024-12-13T14:16:35Z |
| format | Article |
| id | doaj.art-10a5e310a17941dd812473298f28ade0 |
| institution | Directory Open Access Journal |
| issn | 1472-6750 |
| language | English |
| last_indexed | 2024-12-13T14:16:35Z |
| publishDate | 2019-08-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Biotechnology |
| spelling | doaj.art-10a5e310a17941dd812473298f28ade02022-12-21T23:42:17ZengBMCBMC Biotechnology1472-67502019-08-0119111110.1186/s12896-019-0555-1A new primer construction technique that effectively increases amplification of rare mutant templates in samplesJr-Kai Huang0Ling Fan1Tao-Yeuan Wang2Pao-Shu Wu3Department of Pathology, Mackay Memorial HospitalDepartment of Nuclear Medicine, Chang Gung Memorial HospitalDepartment of Pathology, Mackay Memorial HospitalDepartment of Pathology, Mackay Memorial HospitalAbstract Background In personalized medicine, companion diagnostic tests provide additional information to help select a treatment option likely to be optimal for a patient. Although such tests include several techniques for detecting low levels of mutant genes in wild-type backgrounds with fairly high sensitivity, most tests are not specific, and may exhibit high false positive rates. In this study, we describe a new primer structure, named ‘stuntmer’, to selectively suppress amplification of wild-type templates, and promote amplification of mutant templates. Results A single stuntmer for a defined region of DNA can detect several kinds of mutations, including point mutations, deletions, and insertions. Stuntmer PCRs are also highly sensitive, being able to amplify mutant sequences that may make up as little as 0.1% of the DNA sample. Conclusion In conclusion, our technique, stuntmer PCR, can provide a simple, low-cost, highly sensitive, highly accurate, and highly specific platform for developing companion diagnostic tests.http://link.springer.com/article/10.1186/s12896-019-0555-1CancerMutationcfDNA |
| spellingShingle | Jr-Kai Huang Ling Fan Tao-Yeuan Wang Pao-Shu Wu A new primer construction technique that effectively increases amplification of rare mutant templates in samples BMC Biotechnology Cancer Mutation cfDNA |
| title | A new primer construction technique that effectively increases amplification of rare mutant templates in samples |
| title_full | A new primer construction technique that effectively increases amplification of rare mutant templates in samples |
| title_fullStr | A new primer construction technique that effectively increases amplification of rare mutant templates in samples |
| title_full_unstemmed | A new primer construction technique that effectively increases amplification of rare mutant templates in samples |
| title_short | A new primer construction technique that effectively increases amplification of rare mutant templates in samples |
| title_sort | new primer construction technique that effectively increases amplification of rare mutant templates in samples |
| topic | Cancer Mutation cfDNA |
| url | http://link.springer.com/article/10.1186/s12896-019-0555-1 |
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