Target sequence of single cells captured by a polymeric microfluidic device
Abstract Detecting rare circulating tumor cells (CTCs), malignant cells of primary site origin, in the bloodstream is difficult. We previously constructed a polymeric microfluidic device with high capture efficiency for lung cancer cell lines using antibodies against the epithelial cell adhesion mol...
Saved in:
| Main Authors: | , , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2025-08-01
|
| Series: | Scientific Reports |
| Subjects: | |
| Online Access: | https://doi.org/10.1038/s41598-025-14826-y |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849237828283662336 |
|---|---|
| author | Rintaro Oyama Masataka Mori Hiroki Matsumiya Yukiko Nemoto Natsumasa Nishizawa Yohei Honda Masatoshi Kanayama Akihiro Taira Taiji Kuwata Masaru Takenaka Koji Kuroda Kazue Yoneda Takashi Ohnaga Motoyoshi Endo Fumihiro Tanaka |
| author_facet | Rintaro Oyama Masataka Mori Hiroki Matsumiya Yukiko Nemoto Natsumasa Nishizawa Yohei Honda Masatoshi Kanayama Akihiro Taira Taiji Kuwata Masaru Takenaka Koji Kuroda Kazue Yoneda Takashi Ohnaga Motoyoshi Endo Fumihiro Tanaka |
| author_sort | Rintaro Oyama |
| collection | DOAJ |
| description | Abstract Detecting rare circulating tumor cells (CTCs), malignant cells of primary site origin, in the bloodstream is difficult. We previously constructed a polymeric microfluidic device with high capture efficiency for lung cancer cell lines using antibodies against the epithelial cell adhesion molecule (EpCAM). In this study, we investigated a method for extracting DNA from single cells captured in a microfluidic device and performed targeted sequencing using the Cancer Hotspot Panel v2. Additionally, we employed a fixation method, which enabled more efficient sequencing of the EpCAM-chip-captured cells than existing fixation methods. We used blood samples obtained from three patients with lung cancer to assess the clinical applicability of our method. Directly sequenced samples revealed better coverage uniformity than samples subjected to whole-genome sequencing. Direct sequencing of cells fixed with preserver fluid and 100% ethanol was performed accurately with high coverage uniformity. Our method demonstrated a sensitivity of 99.4%, specificity of 99.5%, and area under the curve of 1 (allele frequency cut-off, 18%). Our approach suggests that single cells captured in a microfluidic device are sufficient for genetic mutation analysis. |
| format | Article |
| id | doaj-art-261ab12a3d434374a11fbd88dd2488b3 |
| institution | Kabale University |
| issn | 2045-2322 |
| language | English |
| publishDate | 2025-08-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj-art-261ab12a3d434374a11fbd88dd2488b32025-08-20T04:01:51ZengNature PortfolioScientific Reports2045-23222025-08-0115111310.1038/s41598-025-14826-yTarget sequence of single cells captured by a polymeric microfluidic deviceRintaro Oyama0Masataka Mori1Hiroki Matsumiya2Yukiko Nemoto3Natsumasa Nishizawa4Yohei Honda5Masatoshi Kanayama6Akihiro Taira7Taiji Kuwata8Masaru Takenaka9Koji Kuroda10Kazue Yoneda11Takashi Ohnaga12Motoyoshi Endo13Fumihiro Tanaka14School of Medicine, Second Department of Surgery, University of Occupational and Environmental HealthSchool of Medicine, Second Department of Surgery, University of Occupational and Environmental HealthSchool of Medicine, Second Department of Surgery, University of Occupational and Environmental HealthSchool of Medicine, Second Department of Surgery, University of Occupational and Environmental HealthSchool of Medicine, Second Department of Surgery, University of Occupational and Environmental HealthSchool of Medicine, Second Department of Surgery, University of Occupational and Environmental HealthSchool of Medicine, Second Department of Surgery, University of Occupational and Environmental HealthSchool of Medicine, Second Department of Surgery, University of Occupational and Environmental HealthSchool of Medicine, Second Department of Surgery, University of Occupational and Environmental HealthSchool of Medicine, Second Department of Surgery, University of Occupational and Environmental HealthSchool of Medicine, Second Department of Surgery, University of Occupational and Environmental HealthSchool of Medicine, Second Department of Surgery, University of Occupational and Environmental HealthSchool of Medicine, Second Department of Surgery, University of Occupational and Environmental HealthSchool of Medicine, Department of Molecular Biology, University of Occupational and Environmental HealthSchool of Medicine, Second Department of Surgery, University of Occupational and Environmental HealthAbstract Detecting rare circulating tumor cells (CTCs), malignant cells of primary site origin, in the bloodstream is difficult. We previously constructed a polymeric microfluidic device with high capture efficiency for lung cancer cell lines using antibodies against the epithelial cell adhesion molecule (EpCAM). In this study, we investigated a method for extracting DNA from single cells captured in a microfluidic device and performed targeted sequencing using the Cancer Hotspot Panel v2. Additionally, we employed a fixation method, which enabled more efficient sequencing of the EpCAM-chip-captured cells than existing fixation methods. We used blood samples obtained from three patients with lung cancer to assess the clinical applicability of our method. Directly sequenced samples revealed better coverage uniformity than samples subjected to whole-genome sequencing. Direct sequencing of cells fixed with preserver fluid and 100% ethanol was performed accurately with high coverage uniformity. Our method demonstrated a sensitivity of 99.4%, specificity of 99.5%, and area under the curve of 1 (allele frequency cut-off, 18%). Our approach suggests that single cells captured in a microfluidic device are sufficient for genetic mutation analysis.https://doi.org/10.1038/s41598-025-14826-yCirculating tumor cellCTC-chipPolymeric microfluidic deviceSingle cellTarget sequence |
| spellingShingle | Rintaro Oyama Masataka Mori Hiroki Matsumiya Yukiko Nemoto Natsumasa Nishizawa Yohei Honda Masatoshi Kanayama Akihiro Taira Taiji Kuwata Masaru Takenaka Koji Kuroda Kazue Yoneda Takashi Ohnaga Motoyoshi Endo Fumihiro Tanaka Target sequence of single cells captured by a polymeric microfluidic device Scientific Reports Circulating tumor cell CTC-chip Polymeric microfluidic device Single cell Target sequence |
| title | Target sequence of single cells captured by a polymeric microfluidic device |
| title_full | Target sequence of single cells captured by a polymeric microfluidic device |
| title_fullStr | Target sequence of single cells captured by a polymeric microfluidic device |
| title_full_unstemmed | Target sequence of single cells captured by a polymeric microfluidic device |
| title_short | Target sequence of single cells captured by a polymeric microfluidic device |
| title_sort | target sequence of single cells captured by a polymeric microfluidic device |
| topic | Circulating tumor cell CTC-chip Polymeric microfluidic device Single cell Target sequence |
| url | https://doi.org/10.1038/s41598-025-14826-y |
| work_keys_str_mv | AT rintarooyama targetsequenceofsinglecellscapturedbyapolymericmicrofluidicdevice AT masatakamori targetsequenceofsinglecellscapturedbyapolymericmicrofluidicdevice AT hirokimatsumiya targetsequenceofsinglecellscapturedbyapolymericmicrofluidicdevice AT yukikonemoto targetsequenceofsinglecellscapturedbyapolymericmicrofluidicdevice AT natsumasanishizawa targetsequenceofsinglecellscapturedbyapolymericmicrofluidicdevice AT yoheihonda targetsequenceofsinglecellscapturedbyapolymericmicrofluidicdevice AT masatoshikanayama targetsequenceofsinglecellscapturedbyapolymericmicrofluidicdevice AT akihirotaira targetsequenceofsinglecellscapturedbyapolymericmicrofluidicdevice AT taijikuwata targetsequenceofsinglecellscapturedbyapolymericmicrofluidicdevice AT masarutakenaka targetsequenceofsinglecellscapturedbyapolymericmicrofluidicdevice AT kojikuroda targetsequenceofsinglecellscapturedbyapolymericmicrofluidicdevice AT kazueyoneda targetsequenceofsinglecellscapturedbyapolymericmicrofluidicdevice AT takashiohnaga targetsequenceofsinglecellscapturedbyapolymericmicrofluidicdevice AT motoyoshiendo targetsequenceofsinglecellscapturedbyapolymericmicrofluidicdevice AT fumihirotanaka targetsequenceofsinglecellscapturedbyapolymericmicrofluidicdevice |