Rapid, visual, label-based biosensor platform for identification of hepatitis C virus in clinical applications
Abstract Objectives In the current study, for the first time, we reported a novel HCV molecular diagnostic approach termed reverse transcription loop-mediated isothermal amplification integrated with a gold nanoparticles-based lateral flow biosensor (RT-LAMP-AuNPs-LFB), which we developed for rapid,...
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| Format: | Article |
| Language: | English |
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BMC
2024-02-01
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| Series: | BMC Microbiology |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12866-024-03220-9 |
| _version_ | 1797275660113149952 |
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| author | Yuanfang Shi Qingxue Zhou Shilei Dong Qi Zhao Xue Wu Peng Yang Xiaoyan Zeng Xinggui Yang Yan Tan Xinhua Luo Zhenghua Xiao Xu Chen |
| author_facet | Yuanfang Shi Qingxue Zhou Shilei Dong Qi Zhao Xue Wu Peng Yang Xiaoyan Zeng Xinggui Yang Yan Tan Xinhua Luo Zhenghua Xiao Xu Chen |
| author_sort | Yuanfang Shi |
| collection | DOAJ |
| description | Abstract Objectives In the current study, for the first time, we reported a novel HCV molecular diagnostic approach termed reverse transcription loop-mediated isothermal amplification integrated with a gold nanoparticles-based lateral flow biosensor (RT-LAMP-AuNPs-LFB), which we developed for rapid, sensitive, specific, simple, and visual identification of HCV. Methods A set of LAMP primer was designed according to 5’untranslated region (5’UTR) gene from the major HCV genotypes 1b, 2a, 3b, 6a, and 3a, which are prevalent in China. The HCV-RT-LAMP-AuNPs-LFB assay conditions, including HCV-RT-LAMP reaction temperature and time were optimized. The sensitivity, specificity, and selectivity of our assay were evaluated in the current study. The feasibility of HCV-RT-LAMP-AuNPs-LFB was confirmed through clinical serum samples from patients with suspected HCV infections. Results An unique set of HCV-RT-LAMP primers were successfully designed targeting on the 5’UTR gene. The optimal detection process, including crude nucleic acid extraction (approximately 5 min), RT-LAMP reaction (67℃, 30 min), and visual interpretation of AuNPs-LFB results (~ 2 min), could be performed within 40 min without specific instruments. The limit of detection was determined to be 20 copies per test. The HCV-RT-LAMP-AuNPs-LFB assay exhibited high specificity and anti-interference. Conclusions These preliminary results confirmed that the HCV-RT-LAMP-AuNPs-LFB assay is a sensitive, specific, rapid, visual, and cost-saving assay for identification of HCV. This diagnostic approach has great potential value for point-of-care (POC) diagnostic of HCV, especially in resource-challenged regions. |
| first_indexed | 2024-03-07T15:16:41Z |
| format | Article |
| id | doaj.art-07ec3aa817204459858d834cc365e78e |
| institution | Directory Open Access Journal |
| issn | 1471-2180 |
| language | English |
| last_indexed | 2024-03-07T15:16:41Z |
| publishDate | 2024-02-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Microbiology |
| spelling | doaj.art-07ec3aa817204459858d834cc365e78e2024-03-05T17:51:32ZengBMCBMC Microbiology1471-21802024-02-0124111310.1186/s12866-024-03220-9Rapid, visual, label-based biosensor platform for identification of hepatitis C virus in clinical applicationsYuanfang Shi0Qingxue Zhou1Shilei Dong2Qi Zhao3Xue Wu4Peng Yang5Xiaoyan Zeng6Xinggui Yang7Yan Tan8Xinhua Luo9Zhenghua Xiao10Xu Chen11The Second Clinical Medical College, Guizhou University of Traditional Chinese MedicineClinical Laboratory, Hangzhou Women’s HospitalDepartment of Clinical Laboratory, Zhejiang HospitalDepartment of gastroenterology, the Second Affiliated Hospital, Guizhou University of Traditional Chinese MedicineDepartment of Scientific Research, the Second Affiliated Hospital, Guizhou University of Traditional Chinese MedicineClinical Laboratory, the Second Affiliated Hospital, Guizhou University of Traditional Chinese MedicineCentral Laboratory of the Second Affiliated Hospital, Guizhou University of Traditional Chinese MedicineExperiment Center, Guizhou Provincial Centre for Disease Control and PreventionClinical Laboratory, Guizhou Provincial Center for Clinical LaboratoryDepartment of Infectious Disease, Guizhou Provincial People’s HospitalThe Second Clinical Medical College, Guizhou University of Traditional Chinese MedicineThe Second Clinical Medical College, Guizhou University of Traditional Chinese MedicineAbstract Objectives In the current study, for the first time, we reported a novel HCV molecular diagnostic approach termed reverse transcription loop-mediated isothermal amplification integrated with a gold nanoparticles-based lateral flow biosensor (RT-LAMP-AuNPs-LFB), which we developed for rapid, sensitive, specific, simple, and visual identification of HCV. Methods A set of LAMP primer was designed according to 5’untranslated region (5’UTR) gene from the major HCV genotypes 1b, 2a, 3b, 6a, and 3a, which are prevalent in China. The HCV-RT-LAMP-AuNPs-LFB assay conditions, including HCV-RT-LAMP reaction temperature and time were optimized. The sensitivity, specificity, and selectivity of our assay were evaluated in the current study. The feasibility of HCV-RT-LAMP-AuNPs-LFB was confirmed through clinical serum samples from patients with suspected HCV infections. Results An unique set of HCV-RT-LAMP primers were successfully designed targeting on the 5’UTR gene. The optimal detection process, including crude nucleic acid extraction (approximately 5 min), RT-LAMP reaction (67℃, 30 min), and visual interpretation of AuNPs-LFB results (~ 2 min), could be performed within 40 min without specific instruments. The limit of detection was determined to be 20 copies per test. The HCV-RT-LAMP-AuNPs-LFB assay exhibited high specificity and anti-interference. Conclusions These preliminary results confirmed that the HCV-RT-LAMP-AuNPs-LFB assay is a sensitive, specific, rapid, visual, and cost-saving assay for identification of HCV. This diagnostic approach has great potential value for point-of-care (POC) diagnostic of HCV, especially in resource-challenged regions.https://doi.org/10.1186/s12866-024-03220-9Hepatitis C virusReverse transcription loop-mediated isothermal amplificationBiosensorPoint-of-care platformLimit of detection |
| spellingShingle | Yuanfang Shi Qingxue Zhou Shilei Dong Qi Zhao Xue Wu Peng Yang Xiaoyan Zeng Xinggui Yang Yan Tan Xinhua Luo Zhenghua Xiao Xu Chen Rapid, visual, label-based biosensor platform for identification of hepatitis C virus in clinical applications BMC Microbiology Hepatitis C virus Reverse transcription loop-mediated isothermal amplification Biosensor Point-of-care platform Limit of detection |
| title | Rapid, visual, label-based biosensor platform for identification of hepatitis C virus in clinical applications |
| title_full | Rapid, visual, label-based biosensor platform for identification of hepatitis C virus in clinical applications |
| title_fullStr | Rapid, visual, label-based biosensor platform for identification of hepatitis C virus in clinical applications |
| title_full_unstemmed | Rapid, visual, label-based biosensor platform for identification of hepatitis C virus in clinical applications |
| title_short | Rapid, visual, label-based biosensor platform for identification of hepatitis C virus in clinical applications |
| title_sort | rapid visual label based biosensor platform for identification of hepatitis c virus in clinical applications |
| topic | Hepatitis C virus Reverse transcription loop-mediated isothermal amplification Biosensor Point-of-care platform Limit of detection |
| url | https://doi.org/10.1186/s12866-024-03220-9 |
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