The exploration of droplet digital branched rolling circle amplification based ultrasensitive biosensor for gastric cancer cell-derived extracellular vesicles detection
Techniques for the ultrasensitive detection of cancer cell derived extracellular vesicles (EVs) surface protein are limited. Detection of cancer EVs directly from clinical samples is even more challengeable. Here we developed a droplet digital branched rolling circle amplification (ddBRCA) platform...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-12-01
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| Series: | Materials Today Advances |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2590049822000923 |
| _version_ | 1811201662808752128 |
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| author | R. Huang K. Di K. Adeel B. Fan X. Gu H. Xu H. Shen N. He Z. Li |
| author_facet | R. Huang K. Di K. Adeel B. Fan X. Gu H. Xu H. Shen N. He Z. Li |
| author_sort | R. Huang |
| collection | DOAJ |
| description | Techniques for the ultrasensitive detection of cancer cell derived extracellular vesicles (EVs) surface protein are limited. Detection of cancer EVs directly from clinical samples is even more challengeable. Here we developed a droplet digital branched rolling circle amplification (ddBRCA) platform for the rapid and sensitive detection of cancer EVs. The design includes a hairpin structure DNA containing a cancer EVs specific aptamer sequence and BRCA primer sequence. The presence of target EVs could cause the conformational switching of the hairpin DNA, which subsequently triggers the BRCA. The reaction mixture was encapsulated in nanoliter droplets and allowed to incubate at 30 °C for 45 min. The fluorescent signal of each droplet was then measured using a microchip scanner. The results showed that our platform could detect EVs effectively with a limit of detection down to 12 particles/μL. Furthermore, the platform was tested using serum samples to validate its good prospect for clinical applicability. To sum it up, our platform was the first droplet digital RCA method so far, with advancements such as favorable rapidity (∼2 h), high sensitivity, and no washing step requirement, the proposed biosensor is expected to become a promising tool to be applied in clinical routines to assist in cancer diagnosis. |
| first_indexed | 2024-04-12T02:24:36Z |
| format | Article |
| id | doaj.art-a9de4c1a06984af1aece7c2fef3e91ad |
| institution | Directory Open Access Journal |
| issn | 2590-0498 |
| language | English |
| last_indexed | 2024-04-12T02:24:36Z |
| publishDate | 2022-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Materials Today Advances |
| spelling | doaj.art-a9de4c1a06984af1aece7c2fef3e91ad2022-12-22T03:52:01ZengElsevierMaterials Today Advances2590-04982022-12-0116100296The exploration of droplet digital branched rolling circle amplification based ultrasensitive biosensor for gastric cancer cell-derived extracellular vesicles detectionR. Huang0K. Di1K. Adeel2B. Fan3X. Gu4H. Xu5H. Shen6N. He7Z. Li8Department of Laboratory Medicine, Nanjing University Medical School Affiliated Nanjing Drum Tower Hospital, Nanjing, 210008, China; School of Pharmaceutical Sciences, Nanjing Tech University, Nanjing, 211816, ChinaDepartment of Laboratory Medicine, Nanjing University Medical School Affiliated Nanjing Drum Tower Hospital, Nanjing, 210008, ChinaState Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing, ChinaDepartment of Laboratory Medicine, Nanjing University Medical School Affiliated Nanjing Drum Tower Hospital, Nanjing, 210008, ChinaDepartment of Laboratory Medicine, Nanjing University Medical School Affiliated Nanjing Drum Tower Hospital, Nanjing, 210008, ChinaDepartment of Laboratory Medicine, Nanjing University Medical School Affiliated Nanjing Drum Tower Hospital, Nanjing, 210008, ChinaDepartment of Laboratory Medicine, Nanjing University Medical School Affiliated Nanjing Drum Tower Hospital, Nanjing, 210008, China; Corresponding author.State Key Laboratory of Bioelectronics, School of Biological Science and Medical Engineering, Southeast University, Nanjing, China; Corresponding author.Department of Laboratory Medicine, Nanjing University Medical School Affiliated Nanjing Drum Tower Hospital, Nanjing, 210008, China; Corresponding author.Techniques for the ultrasensitive detection of cancer cell derived extracellular vesicles (EVs) surface protein are limited. Detection of cancer EVs directly from clinical samples is even more challengeable. Here we developed a droplet digital branched rolling circle amplification (ddBRCA) platform for the rapid and sensitive detection of cancer EVs. The design includes a hairpin structure DNA containing a cancer EVs specific aptamer sequence and BRCA primer sequence. The presence of target EVs could cause the conformational switching of the hairpin DNA, which subsequently triggers the BRCA. The reaction mixture was encapsulated in nanoliter droplets and allowed to incubate at 30 °C for 45 min. The fluorescent signal of each droplet was then measured using a microchip scanner. The results showed that our platform could detect EVs effectively with a limit of detection down to 12 particles/μL. Furthermore, the platform was tested using serum samples to validate its good prospect for clinical applicability. To sum it up, our platform was the first droplet digital RCA method so far, with advancements such as favorable rapidity (∼2 h), high sensitivity, and no washing step requirement, the proposed biosensor is expected to become a promising tool to be applied in clinical routines to assist in cancer diagnosis.http://www.sciencedirect.com/science/article/pii/S2590049822000923Extracellular vesiclesDroplet digitalBranched rolling circle amplificationWashing-free |
| spellingShingle | R. Huang K. Di K. Adeel B. Fan X. Gu H. Xu H. Shen N. He Z. Li The exploration of droplet digital branched rolling circle amplification based ultrasensitive biosensor for gastric cancer cell-derived extracellular vesicles detection Materials Today Advances Extracellular vesicles Droplet digital Branched rolling circle amplification Washing-free |
| title | The exploration of droplet digital branched rolling circle amplification based ultrasensitive biosensor for gastric cancer cell-derived extracellular vesicles detection |
| title_full | The exploration of droplet digital branched rolling circle amplification based ultrasensitive biosensor for gastric cancer cell-derived extracellular vesicles detection |
| title_fullStr | The exploration of droplet digital branched rolling circle amplification based ultrasensitive biosensor for gastric cancer cell-derived extracellular vesicles detection |
| title_full_unstemmed | The exploration of droplet digital branched rolling circle amplification based ultrasensitive biosensor for gastric cancer cell-derived extracellular vesicles detection |
| title_short | The exploration of droplet digital branched rolling circle amplification based ultrasensitive biosensor for gastric cancer cell-derived extracellular vesicles detection |
| title_sort | exploration of droplet digital branched rolling circle amplification based ultrasensitive biosensor for gastric cancer cell derived extracellular vesicles detection |
| topic | Extracellular vesicles Droplet digital Branched rolling circle amplification Washing-free |
| url | http://www.sciencedirect.com/science/article/pii/S2590049822000923 |
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