Paper-based ELISA diagnosis technology for human brucellosis based on a multiepitope fusion protein
Brucellosis, as a serious zoonotic infectious disease, has been recognized as a re-emerging disease in the developing countries worldwide. In china, the incidence of brucellosis is increasing each year, seriously threatening the health of humans as well as animal populations. Despite a quite number...
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| creator | Yin, Dehui Bai, Qiongqiong Wu, Xiling Li, Han Shao, Jihong Sun, Mingjun Jiang, Hai Zhang, Jingpeng |
| description | Brucellosis, as a serious zoonotic infectious disease, has been recognized as a re-emerging disease in the developing countries worldwide. In china, the incidence of brucellosis is increasing each year, seriously threatening the health of humans as well as animal populations. Despite a quite number of diagnostic methods currently being used for brucellosis, innovative technologies are still needed for its rapid and accurate diagnosis, especially in area where traditional diagnostic is unavailable.
In this study, a total of 22 B cell linear epitopes were predicted from five Brucella outer membrane proteins (OMPs) using an immunoinformatic approach. These epitopes were then chemically synthesized, and with the method of indirect ELISA (iELISA), each of them displayed a certain degree of capability in identifying human brucellosis positive sera. Subsequently, a fusion protein consisting of the 22 predicted epitopes was prokaryotically expressed and used as diagnostic antigen in a newly established brucellosis testing method, nano-ZnO modified paper-based ELISA (nano-p-ELISA). According to the verifying test using a collection of sera collected from brucellosis and non-brucellosis patients, the sensitivity and specificity of multiepitope based nano-p-ELISA were 92.38% and 98.35% respectively. The positive predictive value was 98.26% and the negative predictive value was 91.67%. The multiepitope based fusion protein also displayed significantly higher specificity than Brucella lipopolysaccharide (LPS) antigen.
B cell epitopes are important candidates for serologically testing brucellosis. Multiepitope fusion protein based nano-p-ELISA displayed significantly sensitivity and specificity compared to Brucella LPS antigen. The strategy applied in this study will be helpful to develop rapid and accurate diagnostic method for brucellosis in human as well as animal populations. |
| doi_str_mv | 10.1371/journal.pntd.0009695 |
| format | Article |
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In this study, a total of 22 B cell linear epitopes were predicted from five Brucella outer membrane proteins (OMPs) using an immunoinformatic approach. These epitopes were then chemically synthesized, and with the method of indirect ELISA (iELISA), each of them displayed a certain degree of capability in identifying human brucellosis positive sera. Subsequently, a fusion protein consisting of the 22 predicted epitopes was prokaryotically expressed and used as diagnostic antigen in a newly established brucellosis testing method, nano-ZnO modified paper-based ELISA (nano-p-ELISA). According to the verifying test using a collection of sera collected from brucellosis and non-brucellosis patients, the sensitivity and specificity of multiepitope based nano-p-ELISA were 92.38% and 98.35% respectively. The positive predictive value was 98.26% and the negative predictive value was 91.67%. The multiepitope based fusion protein also displayed significantly higher specificity than Brucella lipopolysaccharide (LPS) antigen.
B cell epitopes are important candidates for serologically testing brucellosis. Multiepitope fusion protein based nano-p-ELISA displayed significantly sensitivity and specificity compared to Brucella LPS antigen. The strategy applied in this study will be helpful to develop rapid and accurate diagnostic method for brucellosis in human as well as animal populations.</description><identifier>ISSN: 1935-2735</identifier><identifier>ISSN: 1935-2727</identifier><identifier>EISSN: 1935-2735</identifier><identifier>DOI: 10.1371/journal.pntd.0009695</identifier><identifier>PMID: 34403421</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Agglutination tests ; Animal population ; Animal populations ; Antibodies ; Antigenic determinants ; Antigens ; Antigens, Bacterial - analysis ; Antigens, Bacterial - genetics ; Antigens, Bacterial - immunology ; Bacteria ; Bacterial Outer Membrane Proteins - analysis ; Bacterial Outer Membrane Proteins - genetics ; Bacterial Outer Membrane Proteins - immunology ; Biology and Life Sciences ; Biotechnology industry ; Brucella ; Brucella - genetics ; Brucella - immunology ; Brucella - isolation & purification ; Brucellosis ; Brucellosis - diagnosis ; Brucellosis - microbiology ; China ; Developing countries ; Diagnosis ; Diagnostic systems ; Effluents ; ELISA ; Engineering and Technology ; Enzyme-linked immunosorbent assay ; Enzyme-Linked Immunosorbent Assay - instrumentation ; Enzyme-Linked Immunosorbent Assay - methods ; Enzymes ; Epitopes ; Epitopes - analysis ; Epitopes - genetics ; Epitopes - immunology ; Fusion protein ; Health risks ; Humans ; Identification and classification ; Infectious diseases ; Laboratories ; LDCs ; Lipopolysaccharides ; Medical research ; Medicine and Health Sciences ; Membrane proteins ; Methods ; Molecular diagnostic techniques ; Outer membrane proteins ; Packaging ; Peptides ; Populations ; Proteins ; Research and Analysis Methods ; Sensitivity ; Sensitivity and Specificity ; Specificity ; Testing ; Tropical diseases ; Zinc oxide ; Zoonoses</subject><ispartof>PLoS neglected tropical diseases, 2021-08, Vol.15 (8), p.e0009695-e0009695</ispartof><rights>COPYRIGHT 2021 Public Library of Science</rights><rights>2021 Yin et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2021 Yin et al 2021 Yin et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c624t-2ae11c7b9a306c4f39f471c60ab2195ff5628d20cdbe24c6a418c5c73f3764c93</citedby><cites>FETCH-LOGICAL-c624t-2ae11c7b9a306c4f39f471c60ab2195ff5628d20cdbe24c6a418c5c73f3764c93</cites><orcidid>0000-0002-7164-9320</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC8396774/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC8396774/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,724,777,781,861,882,2096,2915,23847,27905,27906,53772,53774,79349,79350</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/34403421$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Barry, Alyssa E</contributor><creatorcontrib>Yin, Dehui</creatorcontrib><creatorcontrib>Bai, Qiongqiong</creatorcontrib><creatorcontrib>Wu, Xiling</creatorcontrib><creatorcontrib>Li, Han</creatorcontrib><creatorcontrib>Shao, Jihong</creatorcontrib><creatorcontrib>Sun, Mingjun</creatorcontrib><creatorcontrib>Jiang, Hai</creatorcontrib><creatorcontrib>Zhang, Jingpeng</creatorcontrib><title>Paper-based ELISA diagnosis technology for human brucellosis based on a multiepitope fusion protein</title><title>PLoS neglected tropical diseases</title><addtitle>PLoS Negl Trop Dis</addtitle><description>Brucellosis, as a serious zoonotic infectious disease, has been recognized as a re-emerging disease in the developing countries worldwide. In china, the incidence of brucellosis is increasing each year, seriously threatening the health of humans as well as animal populations. Despite a quite number of diagnostic methods currently being used for brucellosis, innovative technologies are still needed for its rapid and accurate diagnosis, especially in area where traditional diagnostic is unavailable.
In this study, a total of 22 B cell linear epitopes were predicted from five Brucella outer membrane proteins (OMPs) using an immunoinformatic approach. These epitopes were then chemically synthesized, and with the method of indirect ELISA (iELISA), each of them displayed a certain degree of capability in identifying human brucellosis positive sera. Subsequently, a fusion protein consisting of the 22 predicted epitopes was prokaryotically expressed and used as diagnostic antigen in a newly established brucellosis testing method, nano-ZnO modified paper-based ELISA (nano-p-ELISA). According to the verifying test using a collection of sera collected from brucellosis and non-brucellosis patients, the sensitivity and specificity of multiepitope based nano-p-ELISA were 92.38% and 98.35% respectively. The positive predictive value was 98.26% and the negative predictive value was 91.67%. The multiepitope based fusion protein also displayed significantly higher specificity than Brucella lipopolysaccharide (LPS) antigen.
B cell epitopes are important candidates for serologically testing brucellosis. Multiepitope fusion protein based nano-p-ELISA displayed significantly sensitivity and specificity compared to Brucella LPS antigen. The strategy applied in this study will be helpful to develop rapid and accurate diagnostic method for brucellosis in human as well as animal populations.</description><subject>Agglutination tests</subject><subject>Animal population</subject><subject>Animal populations</subject><subject>Antibodies</subject><subject>Antigenic determinants</subject><subject>Antigens</subject><subject>Antigens, Bacterial - analysis</subject><subject>Antigens, Bacterial - genetics</subject><subject>Antigens, Bacterial - immunology</subject><subject>Bacteria</subject><subject>Bacterial Outer Membrane Proteins - analysis</subject><subject>Bacterial Outer Membrane Proteins - genetics</subject><subject>Bacterial Outer Membrane Proteins - immunology</subject><subject>Biology and Life Sciences</subject><subject>Biotechnology industry</subject><subject>Brucella</subject><subject>Brucella - genetics</subject><subject>Brucella - immunology</subject><subject>Brucella - isolation & purification</subject><subject>Brucellosis</subject><subject>Brucellosis - diagnosis</subject><subject>Brucellosis - microbiology</subject><subject>China</subject><subject>Developing countries</subject><subject>Diagnosis</subject><subject>Diagnostic systems</subject><subject>Effluents</subject><subject>ELISA</subject><subject>Engineering and Technology</subject><subject>Enzyme-linked immunosorbent assay</subject><subject>Enzyme-Linked Immunosorbent Assay - instrumentation</subject><subject>Enzyme-Linked Immunosorbent Assay - methods</subject><subject>Enzymes</subject><subject>Epitopes</subject><subject>Epitopes - analysis</subject><subject>Epitopes - genetics</subject><subject>Epitopes - immunology</subject><subject>Fusion protein</subject><subject>Health risks</subject><subject>Humans</subject><subject>Identification and classification</subject><subject>Infectious diseases</subject><subject>Laboratories</subject><subject>LDCs</subject><subject>Lipopolysaccharides</subject><subject>Medical research</subject><subject>Medicine and Health Sciences</subject><subject>Membrane proteins</subject><subject>Methods</subject><subject>Molecular diagnostic techniques</subject><subject>Outer membrane proteins</subject><subject>Packaging</subject><subject>Peptides</subject><subject>Populations</subject><subject>Proteins</subject><subject>Research and Analysis Methods</subject><subject>Sensitivity</subject><subject>Sensitivity and Specificity</subject><subject>Specificity</subject><subject>Testing</subject><subject>Tropical diseases</subject><subject>Zinc oxide</subject><subject>Zoonoses</subject><issn>1935-2735</issn><issn>1935-2727</issn><issn>1935-2735</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2021</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>DOA</sourceid><recordid>eNptkl2L1DAUhoso7rr6D0QLgngzY74zvVkYllUHBhTU65DmYyZDm9QkFfbfm-50lxlZetFy8rzvOTl9q-otBEuIOfx8CGP0slsOPuslAKBhDX1WXcIG0wXimD4_-b6oXqV0AIA2dAVfVheYEIAJgpeV-iEHExetTEbXt9vNz3Wtndz5kFyqs1F7H7qwu6ttiPV-7KWv2zgq03X3wFEWfC3rfuyyM4PLYTC1HZMr1SGGbJx_Xb2wskvmzfy-qn5_uf11822x_f51c7PeLhRDJC-QNBAq3jYSA6aIxY0lHCoGZItgQ62lDK00Akq3BhHFJIErRRXHFnNGVIOvqvdH36FMJ-b9JIEox4RSxmghNkdCB3kQQ3S9jHciSCfuCyHuhIzZqc4IKK2B1gKglSScsVaRptWtJhZagvXkdT13G9veaGV8jrI7Mz0_8W4vduGvWOGGcU6KwafZIIY_o0lZ9C5Nq5XehHGamyGK4Arjgn74D336djO1k-UCzttQ-qrJVKwZx01JApzaLp-gyqNN71TwxrpSPxN8PBHsjezyPoVuzOUXp3OQHEEVQ0rR2MdlQCCmzD5MLabMijmzRfbudJGPooeQ4n9WNuma</recordid><startdate>20210801</startdate><enddate>20210801</enddate><creator>Yin, Dehui</creator><creator>Bai, Qiongqiong</creator><creator>Wu, Xiling</creator><creator>Li, Han</creator><creator>Shao, Jihong</creator><creator>Sun, Mingjun</creator><creator>Jiang, Hai</creator><creator>Zhang, Jingpeng</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7QL</scope><scope>7SS</scope><scope>7T2</scope><scope>7T7</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8C1</scope><scope>8FD</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BENPR</scope><scope>C1K</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>F1W</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>H94</scope><scope>H95</scope><scope>H97</scope><scope>K9.</scope><scope>L.G</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>P64</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope><orcidid>https://orcid.org/0000-0002-7164-9320</orcidid></search><sort><creationdate>20210801</creationdate><title>Paper-based ELISA diagnosis technology for human brucellosis based on a multiepitope fusion protein</title><author>Yin, Dehui ; Bai, Qiongqiong ; Wu, Xiling ; Li, Han ; Shao, Jihong ; Sun, Mingjun ; Jiang, Hai ; Zhang, Jingpeng</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c624t-2ae11c7b9a306c4f39f471c60ab2195ff5628d20cdbe24c6a418c5c73f3764c93</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2021</creationdate><topic>Agglutination tests</topic><topic>Animal population</topic><topic>Animal populations</topic><topic>Antibodies</topic><topic>Antigenic determinants</topic><topic>Antigens</topic><topic>Antigens, Bacterial - analysis</topic><topic>Antigens, Bacterial - genetics</topic><topic>Antigens, Bacterial - immunology</topic><topic>Bacteria</topic><topic>Bacterial Outer Membrane Proteins - analysis</topic><topic>Bacterial Outer Membrane Proteins - genetics</topic><topic>Bacterial Outer Membrane Proteins - immunology</topic><topic>Biology and Life Sciences</topic><topic>Biotechnology industry</topic><topic>Brucella</topic><topic>Brucella - genetics</topic><topic>Brucella - immunology</topic><topic>Brucella - isolation & purification</topic><topic>Brucellosis</topic><topic>Brucellosis - diagnosis</topic><topic>Brucellosis - microbiology</topic><topic>China</topic><topic>Developing countries</topic><topic>Diagnosis</topic><topic>Diagnostic systems</topic><topic>Effluents</topic><topic>ELISA</topic><topic>Engineering and Technology</topic><topic>Enzyme-linked immunosorbent assay</topic><topic>Enzyme-Linked Immunosorbent Assay - instrumentation</topic><topic>Enzyme-Linked Immunosorbent Assay - methods</topic><topic>Enzymes</topic><topic>Epitopes</topic><topic>Epitopes - 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PLoS neglected tropical diseases</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Yin, Dehui</au><au>Bai, Qiongqiong</au><au>Wu, Xiling</au><au>Li, Han</au><au>Shao, Jihong</au><au>Sun, Mingjun</au><au>Jiang, Hai</au><au>Zhang, Jingpeng</au><au>Barry, Alyssa E</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Paper-based ELISA diagnosis technology for human brucellosis based on a multiepitope fusion protein</atitle><jtitle>PLoS neglected tropical diseases</jtitle><addtitle>PLoS Negl Trop Dis</addtitle><date>2021-08-01</date><risdate>2021</risdate><volume>15</volume><issue>8</issue><spage>e0009695</spage><epage>e0009695</epage><pages>e0009695-e0009695</pages><issn>1935-2735</issn><issn>1935-2727</issn><eissn>1935-2735</eissn><abstract>Brucellosis, as a serious zoonotic infectious disease, has been recognized as a re-emerging disease in the developing countries worldwide. In china, the incidence of brucellosis is increasing each year, seriously threatening the health of humans as well as animal populations. Despite a quite number of diagnostic methods currently being used for brucellosis, innovative technologies are still needed for its rapid and accurate diagnosis, especially in area where traditional diagnostic is unavailable.
In this study, a total of 22 B cell linear epitopes were predicted from five Brucella outer membrane proteins (OMPs) using an immunoinformatic approach. These epitopes were then chemically synthesized, and with the method of indirect ELISA (iELISA), each of them displayed a certain degree of capability in identifying human brucellosis positive sera. Subsequently, a fusion protein consisting of the 22 predicted epitopes was prokaryotically expressed and used as diagnostic antigen in a newly established brucellosis testing method, nano-ZnO modified paper-based ELISA (nano-p-ELISA). According to the verifying test using a collection of sera collected from brucellosis and non-brucellosis patients, the sensitivity and specificity of multiepitope based nano-p-ELISA were 92.38% and 98.35% respectively. The positive predictive value was 98.26% and the negative predictive value was 91.67%. The multiepitope based fusion protein also displayed significantly higher specificity than Brucella lipopolysaccharide (LPS) antigen.
B cell epitopes are important candidates for serologically testing brucellosis. Multiepitope fusion protein based nano-p-ELISA displayed significantly sensitivity and specificity compared to Brucella LPS antigen. The strategy applied in this study will be helpful to develop rapid and accurate diagnostic method for brucellosis in human as well as animal populations.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>34403421</pmid><doi>10.1371/journal.pntd.0009695</doi><orcidid>https://orcid.org/0000-0002-7164-9320</orcidid><oa>free_for_read</oa></addata></record> |
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| identifier | ISSN: 1935-2735 |
| ispartof | PLoS neglected tropical diseases, 2021-08, Vol.15 (8), p.e0009695-e0009695 |
| issn | 1935-2735 1935-2727 1935-2735 |
| language | eng |
| recordid | cdi_gale_infotracmisc_A673927314 |
| source | MEDLINE; DOAJ Directory of Open Access Journals; PubMed Central Open Access; Public Library of Science (PLoS); EZB-FREE-00999 freely available EZB journals; PubMed Central |
| subjects | Agglutination tests Animal population Animal populations Antibodies Antigenic determinants Antigens Antigens, Bacterial - analysis Antigens, Bacterial - genetics Antigens, Bacterial - immunology Bacteria Bacterial Outer Membrane Proteins - analysis Bacterial Outer Membrane Proteins - genetics Bacterial Outer Membrane Proteins - immunology Biology and Life Sciences Biotechnology industry Brucella Brucella - genetics Brucella - immunology Brucella - isolation & purification Brucellosis Brucellosis - diagnosis Brucellosis - microbiology China Developing countries Diagnosis Diagnostic systems Effluents ELISA Engineering and Technology Enzyme-linked immunosorbent assay Enzyme-Linked Immunosorbent Assay - instrumentation Enzyme-Linked Immunosorbent Assay - methods Enzymes Epitopes Epitopes - analysis Epitopes - genetics Epitopes - immunology Fusion protein Health risks Humans Identification and classification Infectious diseases Laboratories LDCs Lipopolysaccharides Medical research Medicine and Health Sciences Membrane proteins Methods Molecular diagnostic techniques Outer membrane proteins Packaging Peptides Populations Proteins Research and Analysis Methods Sensitivity Sensitivity and Specificity Specificity Testing Tropical diseases Zinc oxide Zoonoses |
| title | Paper-based ELISA diagnosis technology for human brucellosis based on a multiepitope fusion protein |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-01-21T04%3A52%3A54IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Paper-based%20ELISA%20diagnosis%20technology%20for%20human%20brucellosis%20based%20on%20a%20multiepitope%20fusion%20protein&rft.jtitle=PLoS%20neglected%20tropical%20diseases&rft.au=Yin,%20Dehui&rft.date=2021-08-01&rft.volume=15&rft.issue=8&rft.spage=e0009695&rft.epage=e0009695&rft.pages=e0009695-e0009695&rft.issn=1935-2735&rft.eissn=1935-2735&rft_id=info:doi/10.1371/journal.pntd.0009695&rft_dat=%3Cgale_plos_%3EA673927314%3C/gale_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=2573455665&rft_id=info:pmid/34403421&rft_galeid=A673927314&rft_doaj_id=oai_doaj_org_article_1afe1ff00dca4766bc49bdbd4f1f43d5&rfr_iscdi=true |