Preservation of cfRNA in cytological supernatants for cfDNA & cfRNA double detection in non‐small cell lung cancer patients
Abstract Backgroud Supernatants from various cytological samples, including body cavity effusion, sputum, bronchoalveolar lavage fluid (BALF), and needle aspiration, have been validated for detecting genetic alterations using cell‐free DNA (cfDNA) in patients with non‐small cell lung cancer (NSCLC)....
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| Format: | Article |
| Language: | English |
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Wiley
2024-09-01
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| Series: | Cancer Medicine |
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| Online Access: | https://doi.org/10.1002/cam4.70197 |
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| author | Yidan Ma Yifei Wang Lei He Jun Du Lin Li Zhixin Bie Yuanming Li Xiaomao Xu Wei Zhou Xiaonan Wu Li Yang Jing Di Chenyang Li Xiaoguang Li Dongge Liu Zheng Wang |
| author_facet | Yidan Ma Yifei Wang Lei He Jun Du Lin Li Zhixin Bie Yuanming Li Xiaomao Xu Wei Zhou Xiaonan Wu Li Yang Jing Di Chenyang Li Xiaoguang Li Dongge Liu Zheng Wang |
| author_sort | Yidan Ma |
| collection | DOAJ |
| description | Abstract Backgroud Supernatants from various cytological samples, including body cavity effusion, sputum, bronchoalveolar lavage fluid (BALF), and needle aspiration, have been validated for detecting genetic alterations using cell‐free DNA (cfDNA) in patients with non‐small cell lung cancer (NSCLC). However, the sensitivity of fusion variations detection remains challenging. The protection of cell‐free RNA (cfRNA) is critical for resolving the issue. Methods A protective solution (PS) was applied for preserving cfRNA in cytological supernatant (CS), and the quality of protected cfRNA was assessed by cycle threshold (CT) values from reverse transcription quantitative polymerase chain reaction (RT‐qPCR). Furthermore, we collected an additional set of malignant cytological and matched tumor samples from 84 NSCLC patients, cfDNA & cfRNA extraction and double detection for driver gene mutations was validated using the multi‐gene mutations detection by RT‐qPCR. Results Under the optimal protection system, 91.0% (101/111) of cfRNA were protected effectively. Among the 84 NSCLC patient samples, seven cytological samples failed the tests. In comparison with tumor samples, the overall sensitivity and specificity of detecting driver genes of supernatant cfDNA and cfRNA were 93.8% (74/77) and 100% (77/77), respectively. Notably, when focusing exclusively on patients with fusion gene changes, both sensitivity and specificity reached 100% (11/11) for EML4‐ALK, ROS1, RET fusions, and MET ex14 skipping. Conclusion These findings suggest that cfDNA & cfRNA extraction and double detection strategy recommended in this study improve the accuracy of driver genes mutations test, especially for RNA‐based assay. |
| format | Article |
| id | doaj-art-d9320a6490a045b5af3bb7c1d5f0811e |
| institution | Kabale University |
| issn | 2045-7634 |
| language | English |
| publishDate | 2024-09-01 |
| publisher | Wiley |
| record_format | Article |
| series | Cancer Medicine |
| spelling | doaj-art-d9320a6490a045b5af3bb7c1d5f0811e2025-02-07T09:08:08ZengWileyCancer Medicine2045-76342024-09-011317n/an/a10.1002/cam4.70197Preservation of cfRNA in cytological supernatants for cfDNA & cfRNA double detection in non‐small cell lung cancer patientsYidan Ma0Yifei Wang1Lei He2Jun Du3Lin Li4Zhixin Bie5Yuanming Li6Xiaomao Xu7Wei Zhou8Xiaonan Wu9Li Yang10Jing Di11Chenyang Li12Xiaoguang Li13Dongge Liu14Zheng Wang15Department of Pathology, Beijing Hospital, National Center of Gerontology Institute of Geriatric Medicine, Chinese Academy of Medical Sciences Beijing People's Republic of ChinaDepartment of Pathology, Beijing Hospital, National Center of Gerontology Institute of Geriatric Medicine, Chinese Academy of Medical Sciences Beijing People's Republic of ChinaDepartment of Pathology, Beijing Hospital, National Center of Gerontology Institute of Geriatric Medicine, Chinese Academy of Medical Sciences Beijing People's Republic of ChinaDepartment of Pathology, Beijing Hospital, National Center of Gerontology Institute of Geriatric Medicine, Chinese Academy of Medical Sciences Beijing People's Republic of ChinaDepartment of Oncology, Beijing Hospital, National Center of Gerontology Institute of Geriatric Medicine, Chinese Academy of Medical Sciences Beijing People's Republic of ChinaDepartment of Minimally Invasive Tumor Therapies Center, Beijing Hospital, National Center of Gerontology Institute of Geriatric Medicine, Chinese Academy of Medical Sciences Beijing People's Republic of ChinaDepartment of Minimally Invasive Tumor Therapies Center, Beijing Hospital, National Center of Gerontology Institute of Geriatric Medicine, Chinese Academy of Medical Sciences Beijing People's Republic of ChinaDepartment of Respiratory and Critical Care Medicine, Beijing Hospital, National Center of Gerontology Institute of Geriatric Medicine, Chinese Academy of Medical Sciences Beijing People's Republic of ChinaDepartment of Respiratory and Critical Care Medicine, Beijing Hospital, National Center of Gerontology Institute of Geriatric Medicine, Chinese Academy of Medical Sciences Beijing People's Republic of ChinaDepartment of Oncology, Beijing Hospital, National Center of Gerontology Institute of Geriatric Medicine, Chinese Academy of Medical Sciences Beijing People's Republic of ChinaDepartment of Pathology, Beijing Hospital, National Center of Gerontology Institute of Geriatric Medicine, Chinese Academy of Medical Sciences Beijing People's Republic of ChinaDepartment of Pathology, Beijing Hospital, National Center of Gerontology Institute of Geriatric Medicine, Chinese Academy of Medical Sciences Beijing People's Republic of ChinaDepartment of Pathology, Beijing Hospital, National Center of Gerontology Institute of Geriatric Medicine, Chinese Academy of Medical Sciences Beijing People's Republic of ChinaDepartment of Minimally Invasive Tumor Therapies Center, Beijing Hospital, National Center of Gerontology Institute of Geriatric Medicine, Chinese Academy of Medical Sciences Beijing People's Republic of ChinaDepartment of Pathology, Beijing Hospital, National Center of Gerontology Institute of Geriatric Medicine, Chinese Academy of Medical Sciences Beijing People's Republic of ChinaDepartment of Pathology, Beijing Hospital, National Center of Gerontology Institute of Geriatric Medicine, Chinese Academy of Medical Sciences Beijing People's Republic of ChinaAbstract Backgroud Supernatants from various cytological samples, including body cavity effusion, sputum, bronchoalveolar lavage fluid (BALF), and needle aspiration, have been validated for detecting genetic alterations using cell‐free DNA (cfDNA) in patients with non‐small cell lung cancer (NSCLC). However, the sensitivity of fusion variations detection remains challenging. The protection of cell‐free RNA (cfRNA) is critical for resolving the issue. Methods A protective solution (PS) was applied for preserving cfRNA in cytological supernatant (CS), and the quality of protected cfRNA was assessed by cycle threshold (CT) values from reverse transcription quantitative polymerase chain reaction (RT‐qPCR). Furthermore, we collected an additional set of malignant cytological and matched tumor samples from 84 NSCLC patients, cfDNA & cfRNA extraction and double detection for driver gene mutations was validated using the multi‐gene mutations detection by RT‐qPCR. Results Under the optimal protection system, 91.0% (101/111) of cfRNA were protected effectively. Among the 84 NSCLC patient samples, seven cytological samples failed the tests. In comparison with tumor samples, the overall sensitivity and specificity of detecting driver genes of supernatant cfDNA and cfRNA were 93.8% (74/77) and 100% (77/77), respectively. Notably, when focusing exclusively on patients with fusion gene changes, both sensitivity and specificity reached 100% (11/11) for EML4‐ALK, ROS1, RET fusions, and MET ex14 skipping. Conclusion These findings suggest that cfDNA & cfRNA extraction and double detection strategy recommended in this study improve the accuracy of driver genes mutations test, especially for RNA‐based assay.https://doi.org/10.1002/cam4.70197cell‐free RNAdriver genefusionliquid biopsyNSCLC |
| spellingShingle | Yidan Ma Yifei Wang Lei He Jun Du Lin Li Zhixin Bie Yuanming Li Xiaomao Xu Wei Zhou Xiaonan Wu Li Yang Jing Di Chenyang Li Xiaoguang Li Dongge Liu Zheng Wang Preservation of cfRNA in cytological supernatants for cfDNA & cfRNA double detection in non‐small cell lung cancer patients Cancer Medicine cell‐free RNA driver gene fusion liquid biopsy NSCLC |
| title | Preservation of cfRNA in cytological supernatants for cfDNA & cfRNA double detection in non‐small cell lung cancer patients |
| title_full | Preservation of cfRNA in cytological supernatants for cfDNA & cfRNA double detection in non‐small cell lung cancer patients |
| title_fullStr | Preservation of cfRNA in cytological supernatants for cfDNA & cfRNA double detection in non‐small cell lung cancer patients |
| title_full_unstemmed | Preservation of cfRNA in cytological supernatants for cfDNA & cfRNA double detection in non‐small cell lung cancer patients |
| title_short | Preservation of cfRNA in cytological supernatants for cfDNA & cfRNA double detection in non‐small cell lung cancer patients |
| title_sort | preservation of cfrna in cytological supernatants for cfdna cfrna double detection in non small cell lung cancer patients |
| topic | cell‐free RNA driver gene fusion liquid biopsy NSCLC |
| url | https://doi.org/10.1002/cam4.70197 |
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