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Rapid and Sensitive Detection of Repetitive Nucleic Acid Sequences Using Magnetically Modulated Biosensors
[Image: see text] Repetitive DNA sequences are abundant in the genome of most biological species. These sequences are naturally “preamplified”, which makes them a preferential target for a variety of biological assays. Current methods to detect specific DNA sequences are based on the quantitative po...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical Society
2019
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6682110/ https://www.ncbi.nlm.nih.gov/pubmed/31460281 http://dx.doi.org/10.1021/acsomega.9b01071 |
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author | Margulis, Michael Danielli, Amos |
author_facet | Margulis, Michael Danielli, Amos |
author_sort | Margulis, Michael |
collection | PubMed |
description | [Image: see text] Repetitive DNA sequences are abundant in the genome of most biological species. These sequences are naturally “preamplified”, which makes them a preferential target for a variety of biological assays. Current methods to detect specific DNA sequences are based on the quantitative polymerase chain reaction (PCR), which relies on target amplification by Taq polymerase and uses a fluorescent resonance energy transfer (FRET)-based probe. Here, to rapidly detect a repetitive DNA sequence, we combine a highly sensitive magnetic modulation biosensing (MMB) system and a modified double-quenched FRET-based probe. The high numbers of copies of the female-specific XhoI sequence of the domestic chicken (Gallus gallus), combined with the low background fluorescence levels of the modified double-quenched probe and the high sensitivity of the MMB system, allow us to determine the chick sex in ovo within 13 min, with 100% sensitivity and specificity. Compared to quantitative PCR, the presented assay is 4–9 times faster. More broadly, by specifically tailoring the primers and probe, the proposed assay can detect any target DNA sequence, either repetitive or nonrepetitive. |
format | Online Article Text |
id | pubmed-6682110 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | American Chemical Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-66821102019-08-27 Rapid and Sensitive Detection of Repetitive Nucleic Acid Sequences Using Magnetically Modulated Biosensors Margulis, Michael Danielli, Amos ACS Omega [Image: see text] Repetitive DNA sequences are abundant in the genome of most biological species. These sequences are naturally “preamplified”, which makes them a preferential target for a variety of biological assays. Current methods to detect specific DNA sequences are based on the quantitative polymerase chain reaction (PCR), which relies on target amplification by Taq polymerase and uses a fluorescent resonance energy transfer (FRET)-based probe. Here, to rapidly detect a repetitive DNA sequence, we combine a highly sensitive magnetic modulation biosensing (MMB) system and a modified double-quenched FRET-based probe. The high numbers of copies of the female-specific XhoI sequence of the domestic chicken (Gallus gallus), combined with the low background fluorescence levels of the modified double-quenched probe and the high sensitivity of the MMB system, allow us to determine the chick sex in ovo within 13 min, with 100% sensitivity and specificity. Compared to quantitative PCR, the presented assay is 4–9 times faster. More broadly, by specifically tailoring the primers and probe, the proposed assay can detect any target DNA sequence, either repetitive or nonrepetitive. American Chemical Society 2019-07-08 /pmc/articles/PMC6682110/ /pubmed/31460281 http://dx.doi.org/10.1021/acsomega.9b01071 Text en Copyright © 2019 American Chemical Society This is an open access article published under an ACS AuthorChoice License (http://pubs.acs.org/page/policy/authorchoice_termsofuse.html) , which permits copying and redistribution of the article or any adaptations for non-commercial purposes. |
spellingShingle | Margulis, Michael Danielli, Amos Rapid and Sensitive Detection of Repetitive Nucleic Acid Sequences Using Magnetically Modulated Biosensors |
title | Rapid and Sensitive Detection of Repetitive Nucleic
Acid Sequences Using Magnetically Modulated Biosensors |
title_full | Rapid and Sensitive Detection of Repetitive Nucleic
Acid Sequences Using Magnetically Modulated Biosensors |
title_fullStr | Rapid and Sensitive Detection of Repetitive Nucleic
Acid Sequences Using Magnetically Modulated Biosensors |
title_full_unstemmed | Rapid and Sensitive Detection of Repetitive Nucleic
Acid Sequences Using Magnetically Modulated Biosensors |
title_short | Rapid and Sensitive Detection of Repetitive Nucleic
Acid Sequences Using Magnetically Modulated Biosensors |
title_sort | rapid and sensitive detection of repetitive nucleic
acid sequences using magnetically modulated biosensors |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6682110/ https://www.ncbi.nlm.nih.gov/pubmed/31460281 http://dx.doi.org/10.1021/acsomega.9b01071 |
work_keys_str_mv | AT margulismichael rapidandsensitivedetectionofrepetitivenucleicacidsequencesusingmagneticallymodulatedbiosensors AT danielliamos rapidandsensitivedetectionofrepetitivenucleicacidsequencesusingmagneticallymodulatedbiosensors |