One-step, rapid, nanoparticle-based biosensor platform for the simultaneous identification of hepatitis B virus and hepatitis C virus in clinical applications
Abstract Objectives Viral hepatitis caused by hepatitis B virus (HBV) and hepatitis C virus (HCV) infections remain a major global public health challenge, particularly in low- and middle-income countries. It is crucial to utilize a pointof-care (POC) testing platform that is sensitive, specific, ra...
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BMC
2024-11-01
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| Series: | BMC Microbiology |
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| Online Access: | https://doi.org/10.1186/s12866-024-03610-z |
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| author | Xu Chen Yuanfang Shi Qi Zhao Yu Wang Xinggui Yang Yan Tan Yi Wang Shilei Dong Zhenghua Xiao |
| author_facet | Xu Chen Yuanfang Shi Qi Zhao Yu Wang Xinggui Yang Yan Tan Yi Wang Shilei Dong Zhenghua Xiao |
| author_sort | Xu Chen |
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| description | Abstract Objectives Viral hepatitis caused by hepatitis B virus (HBV) and hepatitis C virus (HCV) infections remain a major global public health challenge, particularly in low- and middle-income countries. It is crucial to utilize a pointof-care (POC) testing platform that is sensitive, specific, rapid, and user-friendly for screening and diagnosis of the two infections. Here, a novel molecular diagnostic assay, integrating multiplex loop-mediated isothermal amplification with a gold nanoparticle-based lateral flow biosensor (mLAMP-AuNPs-LFB) was developed and applied for one-step, visual, rapid, sensitive, and specific identification of HBV and HCV. Methods The AuNPs-based LFB was devised and constructed for the simultaneous detection of HBV and HCV. The HBV-LAMP and HCV-LAMP primers were designed against the S and 5′-untranslated region (5′-UTR) genes from the major HBV genotypes (B, C, D, B/C recombinant, and C/D recombinant) and HCV subtypes (1b, 2a, 3a, 3b, and 6a) in China, respectively. Our assay conditions, both multiplex-LAMP amplification temperature and time were optimized. The sensitivity and specificity of our assay were tested, and the feasibility of our assay was verified through clinical samples. Results The AuNPs-based LFB used here was successfully manufactured according to our devise manual. The two unique independent primer pairs were successfully designed based on the S and 5′-UTR genes, respectively. The optimal mLAMP-AuNPs-LFB detection process, involving rapid nucleic acid isolation (10 min), mLAMP (63 °C for 35 min), and visual AuNPs-LFB interpretation (less than 2 min), could be completed within 50 min. The HBV&HCV-mLAMP-AuNPs-LFB assay can detect the target genes (HBV-S and HCV-5′-UTR) with as low as 20 copies of plasmid template per test, and the specificity was 100% for the experimental pathogens. Conclusions The preliminary results manifested that our mLAMP-AuNPs-LFB assay is a valuable tool and has tremendous potential as a POC testing approach for HBV and HCV identification, especially in undeveloped regions. |
| format | Article |
| id | doaj-art-b2ee781ea1dd4fc69d0907ae576be0d6 |
| institution | Kabale University |
| issn | 1471-2180 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Microbiology |
| spelling | doaj-art-b2ee781ea1dd4fc69d0907ae576be0d62024-11-10T12:10:52ZengBMCBMC Microbiology1471-21802024-11-0124111310.1186/s12866-024-03610-zOne-step, rapid, nanoparticle-based biosensor platform for the simultaneous identification of hepatitis B virus and hepatitis C virus in clinical applicationsXu Chen0Yuanfang Shi1Qi Zhao2Yu Wang3Xinggui Yang4Yan Tan5Yi Wang6Shilei Dong7Zhenghua Xiao8The Second Clinical Medical College, Guizhou University of Traditional Chinese MedicineThe Second Clinical Medical College, Guizhou University of Traditional Chinese MedicineDepartment of gastroenterology, the Second Affiliated Hospital, Guizhou University of Traditional Chinese MedicineDepartment of Clinical Laboratory Centre, The First People’s Hospital of GuiyangExperimental Center, Guizhou Provincial Centre for Disease Control and PreventionClinical Laboratory, Guizhou Provincial Center for Clinical LaboratoryExperimental Research Center, Capital Institute of PediatricsDepartment of Clinical Laboratory, Zhejiang HospitalThe Second Clinical Medical College, Guizhou University of Traditional Chinese MedicineAbstract Objectives Viral hepatitis caused by hepatitis B virus (HBV) and hepatitis C virus (HCV) infections remain a major global public health challenge, particularly in low- and middle-income countries. It is crucial to utilize a pointof-care (POC) testing platform that is sensitive, specific, rapid, and user-friendly for screening and diagnosis of the two infections. Here, a novel molecular diagnostic assay, integrating multiplex loop-mediated isothermal amplification with a gold nanoparticle-based lateral flow biosensor (mLAMP-AuNPs-LFB) was developed and applied for one-step, visual, rapid, sensitive, and specific identification of HBV and HCV. Methods The AuNPs-based LFB was devised and constructed for the simultaneous detection of HBV and HCV. The HBV-LAMP and HCV-LAMP primers were designed against the S and 5′-untranslated region (5′-UTR) genes from the major HBV genotypes (B, C, D, B/C recombinant, and C/D recombinant) and HCV subtypes (1b, 2a, 3a, 3b, and 6a) in China, respectively. Our assay conditions, both multiplex-LAMP amplification temperature and time were optimized. The sensitivity and specificity of our assay were tested, and the feasibility of our assay was verified through clinical samples. Results The AuNPs-based LFB used here was successfully manufactured according to our devise manual. The two unique independent primer pairs were successfully designed based on the S and 5′-UTR genes, respectively. The optimal mLAMP-AuNPs-LFB detection process, involving rapid nucleic acid isolation (10 min), mLAMP (63 °C for 35 min), and visual AuNPs-LFB interpretation (less than 2 min), could be completed within 50 min. The HBV&HCV-mLAMP-AuNPs-LFB assay can detect the target genes (HBV-S and HCV-5′-UTR) with as low as 20 copies of plasmid template per test, and the specificity was 100% for the experimental pathogens. Conclusions The preliminary results manifested that our mLAMP-AuNPs-LFB assay is a valuable tool and has tremendous potential as a POC testing approach for HBV and HCV identification, especially in undeveloped regions.https://doi.org/10.1186/s12866-024-03610-zHepatitis B virusHepatitis C virusLoop-mediated isothermal amplificationBiosensorPoint-of-care platform |
| spellingShingle | Xu Chen Yuanfang Shi Qi Zhao Yu Wang Xinggui Yang Yan Tan Yi Wang Shilei Dong Zhenghua Xiao One-step, rapid, nanoparticle-based biosensor platform for the simultaneous identification of hepatitis B virus and hepatitis C virus in clinical applications BMC Microbiology Hepatitis B virus Hepatitis C virus Loop-mediated isothermal amplification Biosensor Point-of-care platform |
| title | One-step, rapid, nanoparticle-based biosensor platform for the simultaneous identification of hepatitis B virus and hepatitis C virus in clinical applications |
| title_full | One-step, rapid, nanoparticle-based biosensor platform for the simultaneous identification of hepatitis B virus and hepatitis C virus in clinical applications |
| title_fullStr | One-step, rapid, nanoparticle-based biosensor platform for the simultaneous identification of hepatitis B virus and hepatitis C virus in clinical applications |
| title_full_unstemmed | One-step, rapid, nanoparticle-based biosensor platform for the simultaneous identification of hepatitis B virus and hepatitis C virus in clinical applications |
| title_short | One-step, rapid, nanoparticle-based biosensor platform for the simultaneous identification of hepatitis B virus and hepatitis C virus in clinical applications |
| title_sort | one step rapid nanoparticle based biosensor platform for the simultaneous identification of hepatitis b virus and hepatitis c virus in clinical applications |
| topic | Hepatitis B virus Hepatitis C virus Loop-mediated isothermal amplification Biosensor Point-of-care platform |
| url | https://doi.org/10.1186/s12866-024-03610-z |
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