Multiple cross displacement amplification combined with nanoparticle-based lateral flow biosensor for rapid and sensitive detection of Epstein-Barr virus
IntroductionEpstein-Barr virus (EBV) is a highly dangerous virus that is globally prevalent and closely linked to the development of nasopharyngeal cancer (NPC). Plasma EBV DNA analysis is an effective strategy for early detection, prognostication and monitoring of treatment response of NPC.MethodsH...
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Frontiers Media S.A.
2024-01-01
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| Series: | Frontiers in Cellular and Infection Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcimb.2023.1321394/full |
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| author | Xinbei Jia Juan Zhou Fei Xiao Xiaolan Huang Wenqiang He Wen Hu Yaru Kong Weiheng Yan Jie Ji Yuwei Qi Yi Wang Jun Tai |
| author_facet | Xinbei Jia Juan Zhou Fei Xiao Xiaolan Huang Wenqiang He Wen Hu Yaru Kong Weiheng Yan Jie Ji Yuwei Qi Yi Wang Jun Tai |
| author_sort | Xinbei Jia |
| collection | DOAJ |
| description | IntroductionEpstein-Barr virus (EBV) is a highly dangerous virus that is globally prevalent and closely linked to the development of nasopharyngeal cancer (NPC). Plasma EBV DNA analysis is an effective strategy for early detection, prognostication and monitoring of treatment response of NPC.MethodsHere, we present a novel molecular diagnostic technique termed EBV-MCDA-LFB, which integrates multiple cross displacement amplification (MCDA) with nanoparticle-based lateral flow (LFB) to enable simple, rapid and specific detection of EBV. In the EBV-MCDA-LFB system, a set of 10 primers was designed for rapidly amplifying the highly conserved tandem repeat BamHI-W region of the EBV genome. Subsequently, the LFB facilitate direct assay reading, eliminating the use of extra instruments and reagents.ResultsThe outcomes showed that the 65°C within 40 minutes was the optimal reaction setting for the EBV-MCDA system. The sensitivity of EBV-MCDA-LFB assay reached 7 copies per reaction when using EBV recombinant plasmid, and it showed 100% specificity without any cross-reactivity with other pathogens. The feasibility of the EBV-MCDA-LFB method for EBV detection was successfully validated by 49 clinical plasma samples. The complete detection process, consisting of rapid template extraction (15 minutes), MCDA reaction (65°C for 40 minutes), and LFB result reading (2 minutes), can be finalized within a 60-minutes duration.DiscussionEBV-MCDA-LFB assay designed here is a fast, extremely sensitive and specific technique for detecting EBV in field and at the point-of-care (PoC), which is especially beneficial for countries and regions with a high prevalence of the disease and limited economic resources. |
| first_indexed | 2024-03-08T16:06:32Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 2235-2988 |
| language | English |
| last_indexed | 2024-03-08T16:06:32Z |
| publishDate | 2024-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cellular and Infection Microbiology |
| spelling | doaj.art-85ad0382dbd94c03802f0db81c2bee8f2024-01-08T05:37:54ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882024-01-011310.3389/fcimb.2023.13213941321394Multiple cross displacement amplification combined with nanoparticle-based lateral flow biosensor for rapid and sensitive detection of Epstein-Barr virusXinbei Jia0Juan Zhou1Fei Xiao2Xiaolan Huang3Wenqiang He4Wen Hu5Yaru Kong6Weiheng Yan7Jie Ji8Yuwei Qi9Yi Wang10Jun Tai11Department of Otorhinolaryngology Head and Neck Surgery, Children’s Hospital Capital Institute of Pediatrics, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, ChinaExperimental Research Center, Capital Institute of Pediatrics, Beijing, ChinaExperimental Research Center, Capital Institute of Pediatrics, Beijing, ChinaExperimental Research Center, Capital Institute of Pediatrics, Beijing, ChinaDepartment of Otolaryngology, Head and Neck Surgery, Children’s Hospital Capital Institute of Pediatrics, Beijing, ChinaDepartment of Otolaryngology, Head and Neck Surgery, Children’s Hospital Capital Institute of Pediatrics, Beijing, ChinaDepartment of Otorhinolaryngology Head and Neck Surgery, Children’s Hospital Capital Institute of Pediatrics, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, ChinaDepartment of Otorhinolaryngology Head and Neck Surgery, Children’s Hospital Capital Institute of Pediatrics, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, ChinaDepartment of Otorhinolaryngology Head and Neck Surgery, Beijing Children′s Hospital, Capital Medical University, National Center for Children′s Health, Beijing, ChinaDepartment of Otolaryngology, Head and Neck Surgery, Children’s Hospital Capital Institute of Pediatrics, Beijing, ChinaExperimental Research Center, Capital Institute of Pediatrics, Beijing, ChinaDepartment of Otorhinolaryngology Head and Neck Surgery, Children’s Hospital Capital Institute of Pediatrics, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, ChinaIntroductionEpstein-Barr virus (EBV) is a highly dangerous virus that is globally prevalent and closely linked to the development of nasopharyngeal cancer (NPC). Plasma EBV DNA analysis is an effective strategy for early detection, prognostication and monitoring of treatment response of NPC.MethodsHere, we present a novel molecular diagnostic technique termed EBV-MCDA-LFB, which integrates multiple cross displacement amplification (MCDA) with nanoparticle-based lateral flow (LFB) to enable simple, rapid and specific detection of EBV. In the EBV-MCDA-LFB system, a set of 10 primers was designed for rapidly amplifying the highly conserved tandem repeat BamHI-W region of the EBV genome. Subsequently, the LFB facilitate direct assay reading, eliminating the use of extra instruments and reagents.ResultsThe outcomes showed that the 65°C within 40 minutes was the optimal reaction setting for the EBV-MCDA system. The sensitivity of EBV-MCDA-LFB assay reached 7 copies per reaction when using EBV recombinant plasmid, and it showed 100% specificity without any cross-reactivity with other pathogens. The feasibility of the EBV-MCDA-LFB method for EBV detection was successfully validated by 49 clinical plasma samples. The complete detection process, consisting of rapid template extraction (15 minutes), MCDA reaction (65°C for 40 minutes), and LFB result reading (2 minutes), can be finalized within a 60-minutes duration.DiscussionEBV-MCDA-LFB assay designed here is a fast, extremely sensitive and specific technique for detecting EBV in field and at the point-of-care (PoC), which is especially beneficial for countries and regions with a high prevalence of the disease and limited economic resources.https://www.frontiersin.org/articles/10.3389/fcimb.2023.1321394/fullEpstein-Barr virusmultiple cross displacement amplificationlateral flow biosensorMCDA-LFBnasopharyngeal carcinoma |
| spellingShingle | Xinbei Jia Juan Zhou Fei Xiao Xiaolan Huang Wenqiang He Wen Hu Yaru Kong Weiheng Yan Jie Ji Yuwei Qi Yi Wang Jun Tai Multiple cross displacement amplification combined with nanoparticle-based lateral flow biosensor for rapid and sensitive detection of Epstein-Barr virus Frontiers in Cellular and Infection Microbiology Epstein-Barr virus multiple cross displacement amplification lateral flow biosensor MCDA-LFB nasopharyngeal carcinoma |
| title | Multiple cross displacement amplification combined with nanoparticle-based lateral flow biosensor for rapid and sensitive detection of Epstein-Barr virus |
| title_full | Multiple cross displacement amplification combined with nanoparticle-based lateral flow biosensor for rapid and sensitive detection of Epstein-Barr virus |
| title_fullStr | Multiple cross displacement amplification combined with nanoparticle-based lateral flow biosensor for rapid and sensitive detection of Epstein-Barr virus |
| title_full_unstemmed | Multiple cross displacement amplification combined with nanoparticle-based lateral flow biosensor for rapid and sensitive detection of Epstein-Barr virus |
| title_short | Multiple cross displacement amplification combined with nanoparticle-based lateral flow biosensor for rapid and sensitive detection of Epstein-Barr virus |
| title_sort | multiple cross displacement amplification combined with nanoparticle based lateral flow biosensor for rapid and sensitive detection of epstein barr virus |
| topic | Epstein-Barr virus multiple cross displacement amplification lateral flow biosensor MCDA-LFB nasopharyngeal carcinoma |
| url | https://www.frontiersin.org/articles/10.3389/fcimb.2023.1321394/full |
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