Precision of Genotyping of Epstein-Barr Virus by Polymerase Chain Reaction Using Three Gene Loci (EBNA-2, EBNA-3C, and EBER): Predominance of Type A Virus Associated With Hodgkin’s Disease
To precisely determine the genotype of Epstein-Barr virus (EBV) in Hodgkin’s disease (HD), we simultaneously analyzed three divergent gene loci (EBNA-2, EBNA-3C, and EBER) that distinguish type A and B viruses. The primers designed to amplify these three gene loci encompass either type-specific dele...
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Veröffentlicht in: | Blood 1993-06, Vol.81 (12), p.3372-3381 |
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description | To precisely determine the genotype of Epstein-Barr virus (EBV) in Hodgkin’s disease (HD), we simultaneously analyzed three divergent gene loci (EBNA-2, EBNA-3C, and EBER) that distinguish type A and B viruses. The primers designed to amplify these three gene loci encompass either type-specific deletion sequences (EBNA-2 and EBNA-3C) or type-specific point mutations (EBER) that identify the virus strain based on the sizes of the polymerase chain reaction (PCR)-amplified products or the mobility shifts in single-strand conformation polymorphism analysis. The locations of point mutations were identified by direct sequencing of the PCR-amplified DNA. We analyzed 15 EBV-infected cell lines and found a good correlation between EBNA-2 and EBNA-3C typing results. In contrast, approximately 33% of the cell lines analyzed maintained type A sequences in EBNA-2 and EBNA-3C genes while carrying type B sequences in the EBER region. Data obtained from analysis of cell lines served as a reference for studying HD samples. EBV DNA was detected in about 70% of HD. Among the EBV-positive samples, 56% were associated with type A virus, 13% with type B, and 31 % with dual viral sequences. Thus, type A virus is predominant in HD. Based on the histology, the frequencies of EBV positivity were 83%, 71%, and 33% for mixed cellularity, nodular sclerosis, and lymphocyte predominance, respectively. The detection of high frequency of both type A and B sequences in HD may provide a lead in investigating the role of dual viral infection in EBV pathogenesis. |
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The primers designed to amplify these three gene loci encompass either type-specific deletion sequences (EBNA-2 and EBNA-3C) or type-specific point mutations (EBER) that identify the virus strain based on the sizes of the polymerase chain reaction (PCR)-amplified products or the mobility shifts in single-strand conformation polymorphism analysis. The locations of point mutations were identified by direct sequencing of the PCR-amplified DNA. We analyzed 15 EBV-infected cell lines and found a good correlation between EBNA-2 and EBNA-3C typing results. In contrast, approximately 33% of the cell lines analyzed maintained type A sequences in EBNA-2 and EBNA-3C genes while carrying type B sequences in the EBER region. Data obtained from analysis of cell lines served as a reference for studying HD samples. EBV DNA was detected in about 70% of HD. Among the EBV-positive samples, 56% were associated with type A virus, 13% with type B, and 31 % with dual viral sequences. Thus, type A virus is predominant in HD. Based on the histology, the frequencies of EBV positivity were 83%, 71%, and 33% for mixed cellularity, nodular sclerosis, and lymphocyte predominance, respectively. The detection of high frequency of both type A and B sequences in HD may provide a lead in investigating the role of dual viral infection in EBV pathogenesis.</description><identifier>ISSN: 0006-4971</identifier><identifier>EISSN: 1528-0020</identifier><identifier>DOI: 10.1182/blood.V81.12.3372.3372</identifier><identifier>PMID: 8389616</identifier><language>eng</language><publisher>Washington, DC: Elsevier Inc</publisher><subject>Antigens, Viral - genetics ; Base Sequence ; Biological and medical sciences ; Blotting, Southern ; Cell Line ; DNA-Binding Proteins - genetics ; Epstein-Barr Virus Nuclear Antigens ; Genes, Viral ; Genotype ; Hematologic and hematopoietic diseases ; Herpesvirus 4, Human - genetics ; Hodgkin Disease - microbiology ; Humans ; Leukemias. Malignant lymphomas. Malignant reticulosis. Myelofibrosis ; Medical sciences ; Molecular Sequence Data ; Polymerase Chain Reaction ; Ribosomal Proteins ; RNA-Binding Proteins - genetics</subject><ispartof>Blood, 1993-06, Vol.81 (12), p.3372-3381</ispartof><rights>1993 American Society of Hematology</rights><rights>1993 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c3552-340b04dd13d3e7c8268c0f13c60f6d29f45d1400984eff37e241daf275085b9a3</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=4821338$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8389616$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Lin, Jung-Chung</creatorcontrib><creatorcontrib>Lin, Seh-Ching</creatorcontrib><creatorcontrib>De, Barun K.</creatorcontrib><creatorcontrib>Chan, Weng-Pan</creatorcontrib><creatorcontrib>Evatt, Bruce L.</creatorcontrib><title>Precision of Genotyping of Epstein-Barr Virus by Polymerase Chain Reaction Using Three Gene Loci (EBNA-2, EBNA-3C, and EBER): Predominance of Type A Virus Associated With Hodgkin’s Disease</title><title>Blood</title><addtitle>Blood</addtitle><description>To precisely determine the genotype of Epstein-Barr virus (EBV) in Hodgkin’s disease (HD), we simultaneously analyzed three divergent gene loci (EBNA-2, EBNA-3C, and EBER) that distinguish type A and B viruses. The primers designed to amplify these three gene loci encompass either type-specific deletion sequences (EBNA-2 and EBNA-3C) or type-specific point mutations (EBER) that identify the virus strain based on the sizes of the polymerase chain reaction (PCR)-amplified products or the mobility shifts in single-strand conformation polymorphism analysis. The locations of point mutations were identified by direct sequencing of the PCR-amplified DNA. We analyzed 15 EBV-infected cell lines and found a good correlation between EBNA-2 and EBNA-3C typing results. In contrast, approximately 33% of the cell lines analyzed maintained type A sequences in EBNA-2 and EBNA-3C genes while carrying type B sequences in the EBER region. Data obtained from analysis of cell lines served as a reference for studying HD samples. EBV DNA was detected in about 70% of HD. Among the EBV-positive samples, 56% were associated with type A virus, 13% with type B, and 31 % with dual viral sequences. Thus, type A virus is predominant in HD. Based on the histology, the frequencies of EBV positivity were 83%, 71%, and 33% for mixed cellularity, nodular sclerosis, and lymphocyte predominance, respectively. The detection of high frequency of both type A and B sequences in HD may provide a lead in investigating the role of dual viral infection in EBV pathogenesis.</description><subject>Antigens, Viral - genetics</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Blotting, Southern</subject><subject>Cell Line</subject><subject>DNA-Binding Proteins - genetics</subject><subject>Epstein-Barr Virus Nuclear Antigens</subject><subject>Genes, Viral</subject><subject>Genotype</subject><subject>Hematologic and hematopoietic diseases</subject><subject>Herpesvirus 4, Human - genetics</subject><subject>Hodgkin Disease - microbiology</subject><subject>Humans</subject><subject>Leukemias. Malignant lymphomas. Malignant reticulosis. Myelofibrosis</subject><subject>Medical sciences</subject><subject>Molecular Sequence Data</subject><subject>Polymerase Chain Reaction</subject><subject>Ribosomal Proteins</subject><subject>RNA-Binding Proteins - genetics</subject><issn>0006-4971</issn><issn>1528-0020</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1993</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFUduO0zAQtRBoKQufAPIDQiBtii-5ODzRLWUXqYLVqrs8Wq492RqSONgpUt74DX6Gj-FLcNpoX3mZ0WjOnDM6B6EXlMwpFezttnbOzG8FnVM257w4lgdoRjMmEkIYeYhmhJA8ScuCPkZPQvhGCE05y07QieCizGk-Q3-uPGgbrGuxq_AFtK4fOtvejdOqCz3YNjlX3uNb6_cBbwd85eqhAa8C4OVO2RZfg9L9SHATxsPNzgOMTIDXTlv8enX-eZGwM3zofHmGVWvisLp-8w5HdeMa26pWwyi5GTrAi0lsEUIkUD0Y_NX2O3zpzN132_799TvgDzZAfOEpelSpOsCzqZ-im4-rzfIyWX-5-LRcrBPNs4wlPCVbkhpDueFQaMFyoUlFuc5JlRtWVmlmaEpIKVKoKl4AS6lRFSsyIrJtqfgpenXk7bz7sYfQy8YGDXWtWnD7IIusEDwrWATmR6D2LgQPley8bZQfJCVyDE4egpMxOEmZHDM7lHj4fFLYbxsw92dTUnH_ctqroFVd-WiZDfewVDDKuYiw90cYRDd-WvAyaAvRXWNj0L00zv7vk3_Ap7c-</recordid><startdate>19930615</startdate><enddate>19930615</enddate><creator>Lin, Jung-Chung</creator><creator>Lin, Seh-Ching</creator><creator>De, Barun K.</creator><creator>Chan, Weng-Pan</creator><creator>Evatt, Bruce L.</creator><general>Elsevier Inc</general><general>The Americain Society of Hematology</general><scope>6I.</scope><scope>AAFTH</scope><scope>IQODW</scope><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>7X8</scope></search><sort><creationdate>19930615</creationdate><title>Precision of Genotyping of Epstein-Barr Virus by Polymerase Chain Reaction Using Three Gene Loci (EBNA-2, EBNA-3C, and EBER): Predominance of Type A Virus Associated With Hodgkin’s Disease</title><author>Lin, Jung-Chung ; Lin, Seh-Ching ; De, Barun K. ; Chan, Weng-Pan ; Evatt, Bruce L.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c3552-340b04dd13d3e7c8268c0f13c60f6d29f45d1400984eff37e241daf275085b9a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1993</creationdate><topic>Antigens, Viral - genetics</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Blotting, Southern</topic><topic>Cell Line</topic><topic>DNA-Binding Proteins - genetics</topic><topic>Epstein-Barr Virus Nuclear Antigens</topic><topic>Genes, Viral</topic><topic>Genotype</topic><topic>Hematologic and hematopoietic diseases</topic><topic>Herpesvirus 4, Human - genetics</topic><topic>Hodgkin Disease - microbiology</topic><topic>Humans</topic><topic>Leukemias. Malignant lymphomas. Malignant reticulosis. Myelofibrosis</topic><topic>Medical sciences</topic><topic>Molecular Sequence Data</topic><topic>Polymerase Chain Reaction</topic><topic>Ribosomal Proteins</topic><topic>RNA-Binding Proteins - genetics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Lin, Jung-Chung</creatorcontrib><creatorcontrib>Lin, Seh-Ching</creatorcontrib><creatorcontrib>De, Barun K.</creatorcontrib><creatorcontrib>Chan, Weng-Pan</creatorcontrib><creatorcontrib>Evatt, Bruce L.</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Blood</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Lin, Jung-Chung</au><au>Lin, Seh-Ching</au><au>De, Barun K.</au><au>Chan, Weng-Pan</au><au>Evatt, Bruce L.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Precision of Genotyping of Epstein-Barr Virus by Polymerase Chain Reaction Using Three Gene Loci (EBNA-2, EBNA-3C, and EBER): Predominance of Type A Virus Associated With Hodgkin’s Disease</atitle><jtitle>Blood</jtitle><addtitle>Blood</addtitle><date>1993-06-15</date><risdate>1993</risdate><volume>81</volume><issue>12</issue><spage>3372</spage><epage>3381</epage><pages>3372-3381</pages><issn>0006-4971</issn><eissn>1528-0020</eissn><abstract>To precisely determine the genotype of Epstein-Barr virus (EBV) in Hodgkin’s disease (HD), we simultaneously analyzed three divergent gene loci (EBNA-2, EBNA-3C, and EBER) that distinguish type A and B viruses. The primers designed to amplify these three gene loci encompass either type-specific deletion sequences (EBNA-2 and EBNA-3C) or type-specific point mutations (EBER) that identify the virus strain based on the sizes of the polymerase chain reaction (PCR)-amplified products or the mobility shifts in single-strand conformation polymorphism analysis. The locations of point mutations were identified by direct sequencing of the PCR-amplified DNA. We analyzed 15 EBV-infected cell lines and found a good correlation between EBNA-2 and EBNA-3C typing results. In contrast, approximately 33% of the cell lines analyzed maintained type A sequences in EBNA-2 and EBNA-3C genes while carrying type B sequences in the EBER region. Data obtained from analysis of cell lines served as a reference for studying HD samples. EBV DNA was detected in about 70% of HD. Among the EBV-positive samples, 56% were associated with type A virus, 13% with type B, and 31 % with dual viral sequences. Thus, type A virus is predominant in HD. Based on the histology, the frequencies of EBV positivity were 83%, 71%, and 33% for mixed cellularity, nodular sclerosis, and lymphocyte predominance, respectively. The detection of high frequency of both type A and B sequences in HD may provide a lead in investigating the role of dual viral infection in EBV pathogenesis.</abstract><cop>Washington, DC</cop><pub>Elsevier Inc</pub><pmid>8389616</pmid><doi>10.1182/blood.V81.12.3372.3372</doi><tpages>10</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Antigens, Viral - genetics Base Sequence Biological and medical sciences Blotting, Southern Cell Line DNA-Binding Proteins - genetics Epstein-Barr Virus Nuclear Antigens Genes, Viral Genotype Hematologic and hematopoietic diseases Herpesvirus 4, Human - genetics Hodgkin Disease - microbiology Humans Leukemias. Malignant lymphomas. Malignant reticulosis. Myelofibrosis Medical sciences Molecular Sequence Data Polymerase Chain Reaction Ribosomal Proteins RNA-Binding Proteins - genetics |
title | Precision of Genotyping of Epstein-Barr Virus by Polymerase Chain Reaction Using Three Gene Loci (EBNA-2, EBNA-3C, and EBER): Predominance of Type A Virus Associated With Hodgkin’s Disease |
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