Pulse-Controlled Amplification-A new powerful tool for on-site diagnostics under resource limited conditions.
Background Molecular diagnostics has become essential in the identification of many infectious and neglected diseases, and the detection of nucleic acids often serves as the gold standard technique for most infectious agents. However, established techniques like polymerase chain reaction (PCR) are t...
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ftdoajarticles:oai:doaj.org/article:f142ccb9509a45368bcb8cc65e436cce 2023-05-15T15:13:27+02:00 Pulse-Controlled Amplification-A new powerful tool for on-site diagnostics under resource limited conditions. Katharina Müller Sarah Daßen Scott Holowachuk Katrin Zwirglmaier Joachim Stehr Federico Buersgens Lars Ullerich Kilian Stoecker 2021-01-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0009114 https://doaj.org/article/f142ccb9509a45368bcb8cc65e436cce EN eng Public Library of Science (PLoS) https://doi.org/10.1371/journal.pntd.0009114 https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0009114 https://doaj.org/article/f142ccb9509a45368bcb8cc65e436cce PLoS Neglected Tropical Diseases, Vol 15, Iss 1, p e0009114 (2021) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2021 ftdoajarticles https://doi.org/10.1371/journal.pntd.0009114 2022-12-31T07:48:33Z Background Molecular diagnostics has become essential in the identification of many infectious and neglected diseases, and the detection of nucleic acids often serves as the gold standard technique for most infectious agents. However, established techniques like polymerase chain reaction (PCR) are time-consuming laboratory-bound techniques while rapid tests such as Lateral Flow Immunochromatographic tests often lack the required sensitivity and/or specificity. Methods/principle findings Here we present an affordable, highly mobile alternative method for the rapid identification of infectious agents using pulse-controlled amplification (PCA). PCA is a next generation nucleic acid amplification technology that uses rapid energy pulses to heat microcyclers (micro-scale metal heating elements embedded directly in the amplification reaction) for a few microseconds, thus only heating a small fraction of the reaction volume. The heated microcyclers cool off nearly instantaneously, resulting in ultra-fast heating and cooling cycles during which classic amplification of a target sequence takes place. This reduces the overall amplification time by a factor of up to 10, enabling a sample-to-result workflow in just 15 minutes, while running on a small and portable prototype device. In this proof of principle study, we designed a PCA-assay for the detection of Yersinia pestis to demonstrate the efficacy of this technology. The observed detection limits were 434 copies per reaction (purified DNA) and 35 cells per reaction (crude sample) respectively of Yersinia pestis. Conclusions/significance PCA offers fast and decentralized molecular diagnostics and is applicable whenever rapid, on-site detection of infectious agents is needed, even under resource limited conditions. It combines the sensitivity and specificity of PCR with the rapidness and simplicity of hitherto existing rapid tests. Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLOS Neglected Tropical Diseases 15 1 e0009114 |
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Directory of Open Access Journals: DOAJ Articles |
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English |
topic |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
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Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 Katharina Müller Sarah Daßen Scott Holowachuk Katrin Zwirglmaier Joachim Stehr Federico Buersgens Lars Ullerich Kilian Stoecker Pulse-Controlled Amplification-A new powerful tool for on-site diagnostics under resource limited conditions. |
topic_facet |
Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 |
description |
Background Molecular diagnostics has become essential in the identification of many infectious and neglected diseases, and the detection of nucleic acids often serves as the gold standard technique for most infectious agents. However, established techniques like polymerase chain reaction (PCR) are time-consuming laboratory-bound techniques while rapid tests such as Lateral Flow Immunochromatographic tests often lack the required sensitivity and/or specificity. Methods/principle findings Here we present an affordable, highly mobile alternative method for the rapid identification of infectious agents using pulse-controlled amplification (PCA). PCA is a next generation nucleic acid amplification technology that uses rapid energy pulses to heat microcyclers (micro-scale metal heating elements embedded directly in the amplification reaction) for a few microseconds, thus only heating a small fraction of the reaction volume. The heated microcyclers cool off nearly instantaneously, resulting in ultra-fast heating and cooling cycles during which classic amplification of a target sequence takes place. This reduces the overall amplification time by a factor of up to 10, enabling a sample-to-result workflow in just 15 minutes, while running on a small and portable prototype device. In this proof of principle study, we designed a PCA-assay for the detection of Yersinia pestis to demonstrate the efficacy of this technology. The observed detection limits were 434 copies per reaction (purified DNA) and 35 cells per reaction (crude sample) respectively of Yersinia pestis. Conclusions/significance PCA offers fast and decentralized molecular diagnostics and is applicable whenever rapid, on-site detection of infectious agents is needed, even under resource limited conditions. It combines the sensitivity and specificity of PCR with the rapidness and simplicity of hitherto existing rapid tests. |
format |
Article in Journal/Newspaper |
author |
Katharina Müller Sarah Daßen Scott Holowachuk Katrin Zwirglmaier Joachim Stehr Federico Buersgens Lars Ullerich Kilian Stoecker |
author_facet |
Katharina Müller Sarah Daßen Scott Holowachuk Katrin Zwirglmaier Joachim Stehr Federico Buersgens Lars Ullerich Kilian Stoecker |
author_sort |
Katharina Müller |
title |
Pulse-Controlled Amplification-A new powerful tool for on-site diagnostics under resource limited conditions. |
title_short |
Pulse-Controlled Amplification-A new powerful tool for on-site diagnostics under resource limited conditions. |
title_full |
Pulse-Controlled Amplification-A new powerful tool for on-site diagnostics under resource limited conditions. |
title_fullStr |
Pulse-Controlled Amplification-A new powerful tool for on-site diagnostics under resource limited conditions. |
title_full_unstemmed |
Pulse-Controlled Amplification-A new powerful tool for on-site diagnostics under resource limited conditions. |
title_sort |
pulse-controlled amplification-a new powerful tool for on-site diagnostics under resource limited conditions. |
publisher |
Public Library of Science (PLoS) |
publishDate |
2021 |
url |
https://doi.org/10.1371/journal.pntd.0009114 https://doaj.org/article/f142ccb9509a45368bcb8cc65e436cce |
geographic |
Arctic |
geographic_facet |
Arctic |
genre |
Arctic |
genre_facet |
Arctic |
op_source |
PLoS Neglected Tropical Diseases, Vol 15, Iss 1, p e0009114 (2021) |
op_relation |
https://doi.org/10.1371/journal.pntd.0009114 https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0009114 https://doaj.org/article/f142ccb9509a45368bcb8cc65e436cce |
op_doi |
https://doi.org/10.1371/journal.pntd.0009114 |
container_title |
PLOS Neglected Tropical Diseases |
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15 |
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1 |
container_start_page |
e0009114 |
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1766343996095856640 |