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A modular paper-and-plastic device for tuberculosis nucleic acid amplification testing in limited-resource settings
We present a prototype for conducting rapid, inexpensive and point-of-care-compatible nucleic acid amplification tests (NAATs) for tuberculosis (TB). The fluorescent isothermal paper-and-plastic NAAT (FLIPP-NAAT) uses paper-based loop mediated isothermal amplification (LAMP) for DNA detection. The c...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6814773/ https://www.ncbi.nlm.nih.gov/pubmed/31653930 http://dx.doi.org/10.1038/s41598-019-51873-8 |
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author | Kaur, Navjot Michael, Joy S. Toley, Bhushan J. |
author_facet | Kaur, Navjot Michael, Joy S. Toley, Bhushan J. |
author_sort | Kaur, Navjot |
collection | PubMed |
description | We present a prototype for conducting rapid, inexpensive and point-of-care-compatible nucleic acid amplification tests (NAATs) for tuberculosis (TB). The fluorescent isothermal paper-and-plastic NAAT (FLIPP-NAAT) uses paper-based loop mediated isothermal amplification (LAMP) for DNA detection. The cost of materials required to build a 12-test-zone device is $0.88 and the cost of reagents per reaction is $0.43. An inexpensive imaging platform enables filter-free fluorescence detection of amplified DNA using a cell-phone camera. FLIPP-NAAT can be operated by an untrained user and only requires a regular laboratory incubator as ancillary equipment. All reagents can be dry-stored in the device, facilitating storage and transportation without cold chains. The device design is modular and the assay demonstrated high specificity to Mycobacterium tuberculosis (Mtb), analytical sensitivity of the order of 10 copies of Mtb gDNA, and tolerance to complex samples. The clinical sensitivity and specificity of sputum-based FLIPP NAAT tests were 100% (zero false negatives) and 68.75% (five false positives), respectively (N = 30), using Xpert MTB/RIF assay as the reference standard. FLIPP-NAAT has the potential to provide affordable and accessible molecular diagnostics for TB in low- and middle-income countries, when used in conjunction with an appropriate sample preparation technique. Although demonstrated for the detection of TB, FLIPP-NAAT is a platform technology for amplification of any nucleic acid sequence. |
format | Online Article Text |
id | pubmed-6814773 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-68147732019-10-30 A modular paper-and-plastic device for tuberculosis nucleic acid amplification testing in limited-resource settings Kaur, Navjot Michael, Joy S. Toley, Bhushan J. Sci Rep Article We present a prototype for conducting rapid, inexpensive and point-of-care-compatible nucleic acid amplification tests (NAATs) for tuberculosis (TB). The fluorescent isothermal paper-and-plastic NAAT (FLIPP-NAAT) uses paper-based loop mediated isothermal amplification (LAMP) for DNA detection. The cost of materials required to build a 12-test-zone device is $0.88 and the cost of reagents per reaction is $0.43. An inexpensive imaging platform enables filter-free fluorescence detection of amplified DNA using a cell-phone camera. FLIPP-NAAT can be operated by an untrained user and only requires a regular laboratory incubator as ancillary equipment. All reagents can be dry-stored in the device, facilitating storage and transportation without cold chains. The device design is modular and the assay demonstrated high specificity to Mycobacterium tuberculosis (Mtb), analytical sensitivity of the order of 10 copies of Mtb gDNA, and tolerance to complex samples. The clinical sensitivity and specificity of sputum-based FLIPP NAAT tests were 100% (zero false negatives) and 68.75% (five false positives), respectively (N = 30), using Xpert MTB/RIF assay as the reference standard. FLIPP-NAAT has the potential to provide affordable and accessible molecular diagnostics for TB in low- and middle-income countries, when used in conjunction with an appropriate sample preparation technique. Although demonstrated for the detection of TB, FLIPP-NAAT is a platform technology for amplification of any nucleic acid sequence. Nature Publishing Group UK 2019-10-25 /pmc/articles/PMC6814773/ /pubmed/31653930 http://dx.doi.org/10.1038/s41598-019-51873-8 Text en © The Author(s) 2019 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Kaur, Navjot Michael, Joy S. Toley, Bhushan J. A modular paper-and-plastic device for tuberculosis nucleic acid amplification testing in limited-resource settings |
title | A modular paper-and-plastic device for tuberculosis nucleic acid amplification testing in limited-resource settings |
title_full | A modular paper-and-plastic device for tuberculosis nucleic acid amplification testing in limited-resource settings |
title_fullStr | A modular paper-and-plastic device for tuberculosis nucleic acid amplification testing in limited-resource settings |
title_full_unstemmed | A modular paper-and-plastic device for tuberculosis nucleic acid amplification testing in limited-resource settings |
title_short | A modular paper-and-plastic device for tuberculosis nucleic acid amplification testing in limited-resource settings |
title_sort | modular paper-and-plastic device for tuberculosis nucleic acid amplification testing in limited-resource settings |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6814773/ https://www.ncbi.nlm.nih.gov/pubmed/31653930 http://dx.doi.org/10.1038/s41598-019-51873-8 |
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