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Detection of low-load Epstein-Barr virus in blood samples by enriched recombinase aided amplification assay
Epstein-Barr virus (EBV), a common human γ-herpesvirus, infects more than 90% of adults worldwide. The purpose of this study was to establish a novel EBV detection method by combining the recombinase aided amplification (RAA) assay with an initial enrichment step that utilizes magnetic beads coated...
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| Published in: | AMB Express 2022-06, Vol.12 (1), p.71-71, Article 71 |
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| creator | Li, Jing-yi Chen, Xiao-ping Tie, Yan-qing Sun, Xiu-li Zhang, Rui-qing He, An-na Nie, Ming-zhu Fan, Guo-hao Li, Feng-yu Tian, Feng-yu Shen, Xin-xin Feng, Zhi-shan Ma, Xue-jun |
| description | Epstein-Barr virus (EBV), a common human γ-herpesvirus, infects more than 90% of adults worldwide. The purpose of this study was to establish a novel EBV detection method by combining the recombinase aided amplification (RAA) assay with an initial enrichment step that utilizes magnetic beads coated with a recombinant human mannan-binding lectin (rhMBL, M1 protein). An M1 protein–protein A magnetic bead complex (M1 beads) was prepared and used to achieve separation and enrichment of EBV from blood. After nucleic acid extraction, DNA was amplified by RAA. Using 388 whole blood samples and 1 serum sample, we explored the specificity, sensitivity and applicability of the newly developed detection method and compared it with commercial quantitative real-time polymerase chain reaction (qPCR) following M1 bead enrichment, traditional qPCR and traditional RAA. After enrichment, the positivity rate of EBV was increased from 15.94% to 17.74% by RAA (
P
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| doi_str_mv | 10.1186/s13568-022-01415-9 |
| format | article |
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P
< 0.05) and from 7.20% to 15.17% by qPCR (
P
< 0.05). The viral loads after enrichment were increased by 1.13 to 23.19-fold (
P
< 0.05). Our data demonstrates that an RAA assay incorporating M1 bead enrichment is a promising tool for detecting low EBV viral loads in blood samples that will facilitate an early response to EBV infection.
Graphical abstract
Key points
The RAA with an enrichment step that utilizes magnetic beads coated with M1 protein.
A very effective method for detecting low-load virus in blood samples.
The first report describing virus detection using this method.</description><identifier>ISSN: 2191-0855</identifier><identifier>EISSN: 2191-0855</identifier><identifier>DOI: 10.1186/s13568-022-01415-9</identifier><identifier>PMID: 35689713</identifier><language>eng</language><publisher>Berlin/Heidelberg: Springer Berlin Heidelberg</publisher><subject>Amplification ; Assaying ; Beads ; Biomedical and Life Sciences ; Biotechnology ; Blood ; Enrichment ; Epstein-Barr virus ; Life Sciences ; Low viral load ; M1 beads ; Mannan ; Mannose-binding lectin ; Microbial Genetics and Genomics ; Microbiology ; Nucleic acids ; Original ; Original Article ; Polymerase chain reaction ; Protein A ; Proteins ; RAA ; Recombinase ; Sensitive detection ; Viruses</subject><ispartof>AMB Express, 2022-06, Vol.12 (1), p.71-71, Article 71</ispartof><rights>The Author(s) 2022</rights><rights>2022. The Author(s).</rights><rights>The Author(s) 2022. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c607t-674a9719844e15c44dcee9fc372868a93a1bd9c260f888313506d8e4d6bf0e4f3</citedby><cites>FETCH-LOGICAL-c607t-674a9719844e15c44dcee9fc372868a93a1bd9c260f888313506d8e4d6bf0e4f3</cites><orcidid>0000-0001-5891-5260</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.proquest.com/docview/2675274196/fulltextPDF?pq-origsite=primo$$EPDF$$P50$$Gproquest$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.proquest.com/docview/2675274196?pq-origsite=primo$$EHTML$$P50$$Gproquest$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,885,25753,27924,27925,37012,37013,38516,43895,44590,53791,53793,74412,75126</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/35689713$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Li, Jing-yi</creatorcontrib><creatorcontrib>Chen, Xiao-ping</creatorcontrib><creatorcontrib>Tie, Yan-qing</creatorcontrib><creatorcontrib>Sun, Xiu-li</creatorcontrib><creatorcontrib>Zhang, Rui-qing</creatorcontrib><creatorcontrib>He, An-na</creatorcontrib><creatorcontrib>Nie, Ming-zhu</creatorcontrib><creatorcontrib>Fan, Guo-hao</creatorcontrib><creatorcontrib>Li, Feng-yu</creatorcontrib><creatorcontrib>Tian, Feng-yu</creatorcontrib><creatorcontrib>Shen, Xin-xin</creatorcontrib><creatorcontrib>Feng, Zhi-shan</creatorcontrib><creatorcontrib>Ma, Xue-jun</creatorcontrib><title>Detection of low-load Epstein-Barr virus in blood samples by enriched recombinase aided amplification assay</title><title>AMB Express</title><addtitle>AMB Expr</addtitle><addtitle>AMB Express</addtitle><description>Epstein-Barr virus (EBV), a common human γ-herpesvirus, infects more than 90% of adults worldwide. The purpose of this study was to establish a novel EBV detection method by combining the recombinase aided amplification (RAA) assay with an initial enrichment step that utilizes magnetic beads coated with a recombinant human mannan-binding lectin (rhMBL, M1 protein). An M1 protein–protein A magnetic bead complex (M1 beads) was prepared and used to achieve separation and enrichment of EBV from blood. After nucleic acid extraction, DNA was amplified by RAA. Using 388 whole blood samples and 1 serum sample, we explored the specificity, sensitivity and applicability of the newly developed detection method and compared it with commercial quantitative real-time polymerase chain reaction (qPCR) following M1 bead enrichment, traditional qPCR and traditional RAA. After enrichment, the positivity rate of EBV was increased from 15.94% to 17.74% by RAA (
P
< 0.05) and from 7.20% to 15.17% by qPCR (
P
< 0.05). The viral loads after enrichment were increased by 1.13 to 23.19-fold (
P
< 0.05). Our data demonstrates that an RAA assay incorporating M1 bead enrichment is a promising tool for detecting low EBV viral loads in blood samples that will facilitate an early response to EBV infection.
Graphical abstract
Key points
The RAA with an enrichment step that utilizes magnetic beads coated with M1 protein.
A very effective method for detecting low-load virus in blood samples.
The first report describing virus detection using this method.</description><subject>Amplification</subject><subject>Assaying</subject><subject>Beads</subject><subject>Biomedical and Life Sciences</subject><subject>Biotechnology</subject><subject>Blood</subject><subject>Enrichment</subject><subject>Epstein-Barr virus</subject><subject>Life Sciences</subject><subject>Low viral load</subject><subject>M1 beads</subject><subject>Mannan</subject><subject>Mannose-binding lectin</subject><subject>Microbial Genetics and Genomics</subject><subject>Microbiology</subject><subject>Nucleic acids</subject><subject>Original</subject><subject>Original Article</subject><subject>Polymerase chain reaction</subject><subject>Protein A</subject><subject>Proteins</subject><subject>RAA</subject><subject>Recombinase</subject><subject>Sensitive detection</subject><subject>Viruses</subject><issn>2191-0855</issn><issn>2191-0855</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2022</creationdate><recordtype>article</recordtype><sourceid>COVID</sourceid><sourceid>PIMPY</sourceid><sourceid>DOA</sourceid><recordid>eNp9kkFvFCEUx4nR2GbtF_BgSLx4GQWGYeBiorVqkyZe9EwYeGxZZ2GFmTb77WV3am09yAHIe7_35_HyR-glJW8pleJdoW0nZEMYawjltGvUE3TKqKINkV339MH9BJ2VsiF1dYQo0T1HJ4dS1dP2FP38BBPYKaSIk8djum3GZBy-2JUJQmw-mpzxTchzwSHiYUzJ4WK2uxEKHvYYYg72GhzOYNN2CNEUwCa4GjlAwQdrjtqmFLN_gZ55MxY4uztX6Mfni-_nX5urb18uzz9cNVaQfmpEz01tTknOgXaWc2cBlLdtz6SQRrWGDk5ZJoiXUrZ1DkQ4CdyJwRPgvl2hy0XXJbPRuxy2Ju91MkEfAymvtclTsCNoYrnvHDN1pxwEDJb3HoxTrR96pUjVer9o7eZhC7WTOGUzPhJ9nInhWq_TjVZUSlGbW6E3dwI5_ZqhTHobioVxNBHSXDQTfScIZYpX9PU_6CbNOdZRHSnWc6pEpdhC2ZxKyeDvm6FEH6yhF2voag19tIZWtejVw2_cl_wxQgXaBSg1FdeQ_779H9nfhq7Fdg</recordid><startdate>20220611</startdate><enddate>20220611</enddate><creator>Li, Jing-yi</creator><creator>Chen, Xiao-ping</creator><creator>Tie, Yan-qing</creator><creator>Sun, Xiu-li</creator><creator>Zhang, Rui-qing</creator><creator>He, An-na</creator><creator>Nie, Ming-zhu</creator><creator>Fan, Guo-hao</creator><creator>Li, Feng-yu</creator><creator>Tian, Feng-yu</creator><creator>Shen, Xin-xin</creator><creator>Feng, Zhi-shan</creator><creator>Ma, Xue-jun</creator><general>Springer Berlin Heidelberg</general><general>Springer Nature B.V</general><general>SpringerOpen</general><scope>C6C</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>COVID</scope><scope>DWQXO</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>L6V</scope><scope>LK8</scope><scope>M7P</scope><scope>M7S</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>PTHSS</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope><orcidid>https://orcid.org/0000-0001-5891-5260</orcidid></search><sort><creationdate>20220611</creationdate><title>Detection of low-load Epstein-Barr virus in blood samples by enriched recombinase aided amplification assay</title><author>Li, Jing-yi ; Chen, Xiao-ping ; Tie, Yan-qing ; Sun, Xiu-li ; Zhang, Rui-qing ; He, An-na ; Nie, Ming-zhu ; Fan, Guo-hao ; Li, Feng-yu ; Tian, Feng-yu ; Shen, Xin-xin ; Feng, Zhi-shan ; Ma, Xue-jun</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c607t-674a9719844e15c44dcee9fc372868a93a1bd9c260f888313506d8e4d6bf0e4f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2022</creationdate><topic>Amplification</topic><topic>Assaying</topic><topic>Beads</topic><topic>Biomedical and Life Sciences</topic><topic>Biotechnology</topic><topic>Blood</topic><topic>Enrichment</topic><topic>Epstein-Barr virus</topic><topic>Life Sciences</topic><topic>Low viral load</topic><topic>M1 beads</topic><topic>Mannan</topic><topic>Mannose-binding lectin</topic><topic>Microbial Genetics and Genomics</topic><topic>Microbiology</topic><topic>Nucleic acids</topic><topic>Original</topic><topic>Original Article</topic><topic>Polymerase chain reaction</topic><topic>Protein A</topic><topic>Proteins</topic><topic>RAA</topic><topic>Recombinase</topic><topic>Sensitive detection</topic><topic>Viruses</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Li, Jing-yi</creatorcontrib><creatorcontrib>Chen, Xiao-ping</creatorcontrib><creatorcontrib>Tie, Yan-qing</creatorcontrib><creatorcontrib>Sun, Xiu-li</creatorcontrib><creatorcontrib>Zhang, Rui-qing</creatorcontrib><creatorcontrib>He, An-na</creatorcontrib><creatorcontrib>Nie, Ming-zhu</creatorcontrib><creatorcontrib>Fan, Guo-hao</creatorcontrib><creatorcontrib>Li, Feng-yu</creatorcontrib><creatorcontrib>Tian, Feng-yu</creatorcontrib><creatorcontrib>Shen, Xin-xin</creatorcontrib><creatorcontrib>Feng, Zhi-shan</creatorcontrib><creatorcontrib>Ma, Xue-jun</creatorcontrib><collection>SpringerOpen</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Materials Science & Engineering Collection</collection><collection>ProQuest Central (Alumni)</collection><collection>ProQuest Central</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>AUTh Library subscriptions: ProQuest Central</collection><collection>Technology Collection</collection><collection>Natural Science Collection</collection><collection>ProQuest One Community College</collection><collection>Coronavirus Research Database</collection><collection>ProQuest Central Korea</collection><collection>ProQuest Central Student</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Engineering Collection</collection><collection>ProQuest Biological Science Collection</collection><collection>Biological Science Database</collection><collection>Engineering Database</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>Engineering Collection</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>AMB Express</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Li, Jing-yi</au><au>Chen, Xiao-ping</au><au>Tie, Yan-qing</au><au>Sun, Xiu-li</au><au>Zhang, Rui-qing</au><au>He, An-na</au><au>Nie, Ming-zhu</au><au>Fan, Guo-hao</au><au>Li, Feng-yu</au><au>Tian, Feng-yu</au><au>Shen, Xin-xin</au><au>Feng, Zhi-shan</au><au>Ma, Xue-jun</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Detection of low-load Epstein-Barr virus in blood samples by enriched recombinase aided amplification assay</atitle><jtitle>AMB Express</jtitle><stitle>AMB Expr</stitle><addtitle>AMB Express</addtitle><date>2022-06-11</date><risdate>2022</risdate><volume>12</volume><issue>1</issue><spage>71</spage><epage>71</epage><pages>71-71</pages><artnum>71</artnum><issn>2191-0855</issn><eissn>2191-0855</eissn><abstract>Epstein-Barr virus (EBV), a common human γ-herpesvirus, infects more than 90% of adults worldwide. The purpose of this study was to establish a novel EBV detection method by combining the recombinase aided amplification (RAA) assay with an initial enrichment step that utilizes magnetic beads coated with a recombinant human mannan-binding lectin (rhMBL, M1 protein). An M1 protein–protein A magnetic bead complex (M1 beads) was prepared and used to achieve separation and enrichment of EBV from blood. After nucleic acid extraction, DNA was amplified by RAA. Using 388 whole blood samples and 1 serum sample, we explored the specificity, sensitivity and applicability of the newly developed detection method and compared it with commercial quantitative real-time polymerase chain reaction (qPCR) following M1 bead enrichment, traditional qPCR and traditional RAA. After enrichment, the positivity rate of EBV was increased from 15.94% to 17.74% by RAA (
P
< 0.05) and from 7.20% to 15.17% by qPCR (
P
< 0.05). The viral loads after enrichment were increased by 1.13 to 23.19-fold (
P
< 0.05). Our data demonstrates that an RAA assay incorporating M1 bead enrichment is a promising tool for detecting low EBV viral loads in blood samples that will facilitate an early response to EBV infection.
Graphical abstract
Key points
The RAA with an enrichment step that utilizes magnetic beads coated with M1 protein.
A very effective method for detecting low-load virus in blood samples.
The first report describing virus detection using this method.</abstract><cop>Berlin/Heidelberg</cop><pub>Springer Berlin Heidelberg</pub><pmid>35689713</pmid><doi>10.1186/s13568-022-01415-9</doi><tpages>1</tpages><orcidid>https://orcid.org/0000-0001-5891-5260</orcidid><oa>free_for_read</oa></addata></record> |
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| ispartof | AMB Express, 2022-06, Vol.12 (1), p.71-71, Article 71 |
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| source | Publicly Available Content Database; Springer Nature - SpringerLink Journals - Fully Open Access ; PubMed Central; Free Full-Text Journals in Chemistry; Coronavirus Research Database |
| subjects | Amplification Assaying Beads Biomedical and Life Sciences Biotechnology Blood Enrichment Epstein-Barr virus Life Sciences Low viral load M1 beads Mannan Mannose-binding lectin Microbial Genetics and Genomics Microbiology Nucleic acids Original Original Article Polymerase chain reaction Protein A Proteins RAA Recombinase Sensitive detection Viruses |
| title | Detection of low-load Epstein-Barr virus in blood samples by enriched recombinase aided amplification assay |
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