A novel linear displacement isothermal amplification with strand displacement probes (LDIA-SD) in a pocket-size device for point-of-care testing of infectious diseases
© 2022 Published by Elsevier B.V..
Nucleic acid amplification is crucial for disease diagnosis, especially lethal infectious diseases such as COVID-19. Compared with PCR, isothermal amplification methods are advantageous for point-of-care testing (POCT). However, complicated primer design limits their application in detecting some short targets or sequences with abnormal GC content. Herein, we developed a novel linear displacement isothermal amplification (LDIA) method using two pairs of conventional primers and Bacillus stearothermophilus (Bst) DNA polymerase, and reactions could be accelerated by adding an extra primer. Pseudorabies virus gE (high GC content) and Salmonella fimW (low GC content) genes were used to evaluate the LDIA assay. Using strand displacement (SD) probes, a LDIA-SD method was developed to realize probe-based specific detection. Additionally, we incorporated a nucleic acid-free extraction step and a pocket-sized device to realize POCT applications of the LDIA-SD method. The LDIA-SD method has advantages including facile primer design, high sensitivity and specificity, and applicability for POCT, especially for amplification of complex sequences and detection of infectious diseases.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2023 |
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| Erschienen: |
2023 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:379 |
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| Enthalten in: |
Sensors and actuators. B, Chemical - 379(2023) vom: 15. März, Seite 133244 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Gou, Hongchao [VerfasserIn] |
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| Links: |
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| Themen: |
Infectious diseases |
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| Anmerkungen: |
Date Revised 03.01.2023 published: Print-Electronic Citation Status PubMed-not-MEDLINE |
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| doi: |
10.1016/j.snb.2022.133244 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
NLM351012176 |
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| 500 | |a Citation Status PubMed-not-MEDLINE | ||
| 520 | |a © 2022 Published by Elsevier B.V. | ||
| 520 | |a Nucleic acid amplification is crucial for disease diagnosis, especially lethal infectious diseases such as COVID-19. Compared with PCR, isothermal amplification methods are advantageous for point-of-care testing (POCT). However, complicated primer design limits their application in detecting some short targets or sequences with abnormal GC content. Herein, we developed a novel linear displacement isothermal amplification (LDIA) method using two pairs of conventional primers and Bacillus stearothermophilus (Bst) DNA polymerase, and reactions could be accelerated by adding an extra primer. Pseudorabies virus gE (high GC content) and Salmonella fimW (low GC content) genes were used to evaluate the LDIA assay. Using strand displacement (SD) probes, a LDIA-SD method was developed to realize probe-based specific detection. Additionally, we incorporated a nucleic acid-free extraction step and a pocket-sized device to realize POCT applications of the LDIA-SD method. The LDIA-SD method has advantages including facile primer design, high sensitivity and specificity, and applicability for POCT, especially for amplification of complex sequences and detection of infectious diseases | ||
| 650 | 4 | |a Journal Article | |
| 650 | 4 | |a Infectious diseases | |
| 650 | 4 | |a Isothermal amplification | |
| 650 | 4 | |a Linear displacement isothermal amplification (LDIA) | |
| 650 | 4 | |a Nucleic Acid Amplification | |
| 650 | 4 | |a Point-of-care testing (POCT) | |
| 700 | 1 | |a Lin, Qijie |e verfasserin |4 aut | |
| 700 | 1 | |a Shen, Haiyan |e verfasserin |4 aut | |
| 700 | 1 | |a Jia, Kaiyuan |e verfasserin |4 aut | |
| 700 | 1 | |a Liang, Yucen |e verfasserin |4 aut | |
| 700 | 1 | |a Peng, Junhao |e verfasserin |4 aut | |
| 700 | 1 | |a Zhang, Chunhong |e verfasserin |4 aut | |
| 700 | 1 | |a Qu, Xiaoyun |e verfasserin |4 aut | |
| 700 | 1 | |a Li, Yanbin |e verfasserin |4 aut | |
| 700 | 1 | |a Lin, Jianhan |e verfasserin |4 aut | |
| 700 | 1 | |a Zhang, Jianmin |e verfasserin |4 aut | |
| 700 | 1 | |a Liao, Ming |e verfasserin |4 aut | |
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