Fluorescence microarrays for enzyme-free DNA detection based on web hybrid chain reaction
The diagnosis of pathogenic microorganism infection mainly relies on molecular detection of nucleic acids or proteins, and the nucleic acid detection is the gold standard for the pathogen detection. The viral genome mutates quickly, and the genome of the host cell usually binds to multiple sites in...
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| Format: | Article |
| Language: | English |
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Elsevier
2022-09-01
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| Series: | Biosensors and Bioelectronics: X |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2590137022000462 |
| _version_ | 1811183269462409216 |
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| author | Furui Jin La Ta Min Liu Yanyan Sun Yipu Pan Zhoumin Li Danke Xu |
| author_facet | Furui Jin La Ta Min Liu Yanyan Sun Yipu Pan Zhoumin Li Danke Xu |
| author_sort | Furui Jin |
| collection | DOAJ |
| description | The diagnosis of pathogenic microorganism infection mainly relies on molecular detection of nucleic acids or proteins, and the nucleic acid detection is the gold standard for the pathogen detection. The viral genome mutates quickly, and the genome of the host cell usually binds to multiple sites in the viral genome, leading to patient infections. Therefore, rapid nucleic acid detection methods with high specificity are essential for controlling the spread of viruses and maintaining human health. In this study, Cy3-labeled DNA probes were obtained based on a convenient and enzyme-free web hybridization chain reaction (wHCR) and used for DNA diagnosis of hepatitis B virus (HBV) in an isothermal platform. In this microarray platform, the target DNA is directly captured on the microarray and identified by the aggregated DNA probes to amplify the fluorescence signal. After fluorescence scanning analysis, the detection limit of HBV DNA fragments on this DNA microarray is 84 fM. In addition, this method not only specifically distinguishes single-base mismatched sequences, but also obtains the quantitative detection of HBV DNA in serum samples. Compared with the enzymatic amplification reaction in which the target nucleic acids are used as the amplification template, this method effectively avoids cross-contamination. Furthermore, compared to enzymatic reactions, the requirements for reaction conditions such as temperature and pH are relatively lenient. This method is user-friendly and suitable for molecular detection of infectious diseases under limited resource conditions. |
| first_indexed | 2024-04-11T09:45:14Z |
| format | Article |
| id | doaj.art-4370c073c9404b3da880f36b6fc6b89e |
| institution | Directory Open Access Journal |
| issn | 2590-1370 |
| language | English |
| last_indexed | 2024-04-11T09:45:14Z |
| publishDate | 2022-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Biosensors and Bioelectronics: X |
| spelling | doaj.art-4370c073c9404b3da880f36b6fc6b89e2022-12-22T04:31:05ZengElsevierBiosensors and Bioelectronics: X2590-13702022-09-0111100151Fluorescence microarrays for enzyme-free DNA detection based on web hybrid chain reactionFurui Jin0La Ta1Min Liu2Yanyan Sun3Yipu Pan4Zhoumin Li5Danke Xu6State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, 210023, China; Tianjin Institute for Food Safety Inspection Technology, Tianjin, 300308, ChinaState Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, 210023, ChinaState Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, 210023, ChinaState Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, 210023, ChinaState Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, 210023, ChinaNanjing University Jinling College, Nanjing, 210089, China; Nanjing Xiangzhong Biological Technology Co., Ltd, Nanjing, 210022, China; Corresponding author. Nanjing University Jinling College, Nanjing, 210089, China.State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University, Nanjing, 210023, China; Corresponding author.The diagnosis of pathogenic microorganism infection mainly relies on molecular detection of nucleic acids or proteins, and the nucleic acid detection is the gold standard for the pathogen detection. The viral genome mutates quickly, and the genome of the host cell usually binds to multiple sites in the viral genome, leading to patient infections. Therefore, rapid nucleic acid detection methods with high specificity are essential for controlling the spread of viruses and maintaining human health. In this study, Cy3-labeled DNA probes were obtained based on a convenient and enzyme-free web hybridization chain reaction (wHCR) and used for DNA diagnosis of hepatitis B virus (HBV) in an isothermal platform. In this microarray platform, the target DNA is directly captured on the microarray and identified by the aggregated DNA probes to amplify the fluorescence signal. After fluorescence scanning analysis, the detection limit of HBV DNA fragments on this DNA microarray is 84 fM. In addition, this method not only specifically distinguishes single-base mismatched sequences, but also obtains the quantitative detection of HBV DNA in serum samples. Compared with the enzymatic amplification reaction in which the target nucleic acids are used as the amplification template, this method effectively avoids cross-contamination. Furthermore, compared to enzymatic reactions, the requirements for reaction conditions such as temperature and pH are relatively lenient. This method is user-friendly and suitable for molecular detection of infectious diseases under limited resource conditions.http://www.sciencedirect.com/science/article/pii/S2590137022000462MicroarrayFluorescence enhancementEnzyme-freeHepatitis B virusHybridization chain reaction |
| spellingShingle | Furui Jin La Ta Min Liu Yanyan Sun Yipu Pan Zhoumin Li Danke Xu Fluorescence microarrays for enzyme-free DNA detection based on web hybrid chain reaction Biosensors and Bioelectronics: X Microarray Fluorescence enhancement Enzyme-free Hepatitis B virus Hybridization chain reaction |
| title | Fluorescence microarrays for enzyme-free DNA detection based on web hybrid chain reaction |
| title_full | Fluorescence microarrays for enzyme-free DNA detection based on web hybrid chain reaction |
| title_fullStr | Fluorescence microarrays for enzyme-free DNA detection based on web hybrid chain reaction |
| title_full_unstemmed | Fluorescence microarrays for enzyme-free DNA detection based on web hybrid chain reaction |
| title_short | Fluorescence microarrays for enzyme-free DNA detection based on web hybrid chain reaction |
| title_sort | fluorescence microarrays for enzyme free dna detection based on web hybrid chain reaction |
| topic | Microarray Fluorescence enhancement Enzyme-free Hepatitis B virus Hybridization chain reaction |
| url | http://www.sciencedirect.com/science/article/pii/S2590137022000462 |
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