Rapid technique of DNA-DNA in situ hybridisation on formalin fixed tissue sections using microwave irradiation
The relative sensitivities of different protocols for detecting cytomegalovirus nucleic acid sequences in histological specimens, using a biotinylated cDNA probe, were assessed. Several commonly used pre-treatment steps were not essential, nor was the use of a highly sensitive detection system. The...
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| Veröffentlicht in: | Journal of clinical pathology 1987-08, Vol.40 (8), p.865-869 |
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| container_end_page | 869 |
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| container_issue | 8 |
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| container_title | Journal of clinical pathology |
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| creator | Coates, P J Hall, P A Butler, M G D'Ardenne, A J |
| description | The relative sensitivities of different protocols for detecting cytomegalovirus nucleic acid sequences in histological specimens, using a biotinylated cDNA probe, were assessed. Several commonly used pre-treatment steps were not essential, nor was the use of a highly sensitive detection system. The choice of enzyme used for proteolytic digestion of tissue seems to be important, and increasing the temperature of denaturation of tissue and probe DNA to above 100 degrees C greatly increased the sensitivity of the method. Difficulties in achieving such high temperatures in a controlled manner were overcome by the use of a rapid microwave heating method that can be used routinely in laboratories. This technique detected cytomegalovirus infections in formalin fixed, paraffin processed tissue sections within a single working day. |
| doi_str_mv | 10.1136/jcp.40.8.865 |
| format | Article |
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Several commonly used pre-treatment steps were not essential, nor was the use of a highly sensitive detection system. The choice of enzyme used for proteolytic digestion of tissue seems to be important, and increasing the temperature of denaturation of tissue and probe DNA to above 100 degrees C greatly increased the sensitivity of the method. Difficulties in achieving such high temperatures in a controlled manner were overcome by the use of a rapid microwave heating method that can be used routinely in laboratories. This technique detected cytomegalovirus infections in formalin fixed, paraffin processed tissue sections within a single working day.</description><identifier>ISSN: 0021-9746</identifier><identifier>EISSN: 1472-4146</identifier><identifier>DOI: 10.1136/jcp.40.8.865</identifier><identifier>PMID: 2821079</identifier><identifier>CODEN: JCPAAK</identifier><language>eng</language><publisher>London: BMJ Publishing Group Ltd and Association of Clinical Pathologists</publisher><subject>Analytical, structural and metabolic biochemistry ; Biological and medical sciences ; Cytomegalovirus - isolation & purification ; Cytomegalovirus Infections - diagnosis ; Cytomegalovirus Infections - microbiology ; Dna, deoxyribonucleoproteins ; DNA, Viral - analysis ; Fundamental and applied biological sciences. Psychology ; Humans ; Microwaves ; Nucleic Acid Denaturation ; Nucleic Acid Hybridization ; Nucleic acids ; Temperature</subject><ispartof>Journal of clinical pathology, 1987-08, Vol.40 (8), p.865-869</ispartof><rights>1988 INIST-CNRS</rights><rights>Copyright BMJ Publishing Group LTD Aug 1987</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-b4215-6efcb9e11a42110841f6fb19a4a3ebe14dcbbd4254431ef8510762b7dcff54323</citedby><cites>FETCH-LOGICAL-b4215-6efcb9e11a42110841f6fb19a4a3ebe14dcbbd4254431ef8510762b7dcff54323</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC1141126/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC1141126/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,727,780,784,885,27923,27924,53790,53792</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=7689435$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/2821079$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Coates, P J</creatorcontrib><creatorcontrib>Hall, P A</creatorcontrib><creatorcontrib>Butler, M G</creatorcontrib><creatorcontrib>D'Ardenne, A J</creatorcontrib><title>Rapid technique of DNA-DNA in situ hybridisation on formalin fixed tissue sections using microwave irradiation</title><title>Journal of clinical pathology</title><addtitle>J Clin Pathol</addtitle><description>The relative sensitivities of different protocols for detecting cytomegalovirus nucleic acid sequences in histological specimens, using a biotinylated cDNA probe, were assessed. Several commonly used pre-treatment steps were not essential, nor was the use of a highly sensitive detection system. The choice of enzyme used for proteolytic digestion of tissue seems to be important, and increasing the temperature of denaturation of tissue and probe DNA to above 100 degrees C greatly increased the sensitivity of the method. Difficulties in achieving such high temperatures in a controlled manner were overcome by the use of a rapid microwave heating method that can be used routinely in laboratories. This technique detected cytomegalovirus infections in formalin fixed, paraffin processed tissue sections within a single working day.</description><subject>Analytical, structural and metabolic biochemistry</subject><subject>Biological and medical sciences</subject><subject>Cytomegalovirus - isolation & purification</subject><subject>Cytomegalovirus Infections - diagnosis</subject><subject>Cytomegalovirus Infections - microbiology</subject><subject>Dna, deoxyribonucleoproteins</subject><subject>DNA, Viral - analysis</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Humans</subject><subject>Microwaves</subject><subject>Nucleic Acid Denaturation</subject><subject>Nucleic Acid Hybridization</subject><subject>Nucleic acids</subject><subject>Temperature</subject><issn>0021-9746</issn><issn>1472-4146</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1987</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>GNUQQ</sourceid><recordid>eNp9kcGLEzEUxoMoa129eRUGFL04NS_JJDMXYalWhd2Koou3kGSSbepMpiYz6-5_b2pLUQ9CQgjf73289z6EHgOeA1D-amO2c4bn9bzm1R00AyZIyYDxu2iGMYGyEYzfRw9S2mAMVAA9QSekJoBFM0Phs9r6thitWQf_Y7LF4Io3q7My38KHIvlxKta3OvrWJzX6IRT5uCH2qsuy8zc2F_uUcmWyZgekYko-XBW9N3H4qa5t4WNUrf9d_RDdc6pL9tHhPUVfl2-_LN6X5x_ffVicnZeaEahKbp3RjQVQ-Qu4ZuC409AopqjVFlhrtG4ZqRijYF1d5WE40aI1zlWMEnqKXu99t5PubWtsGKPq5Db6XsVbOSgv_1aCX8ur4VoCMADCs8Hzg0Ec8lrSKHufjO06FewwJVkDJpxVkMGn_4CbYYohDydBCMxojRuRqZd7Ku8kpWjdsRXAcpeizClKhmUtc4oZf_Jn-0f4EFvWnx10lYzqXFTB-HTEBK8bRnc25R7zabQ3R1nF75ILKiq5ulzI5erbxcWn5aXczfJiz-t-8_8GfwHTrsKS</recordid><startdate>19870801</startdate><enddate>19870801</enddate><creator>Coates, P J</creator><creator>Hall, P A</creator><creator>Butler, M G</creator><creator>D'Ardenne, A J</creator><general>BMJ Publishing Group Ltd and Association of Clinical Pathologists</general><general>BMJ</general><general>BMJ Publishing Group LTD</general><scope>BSCLL</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>3V.</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>88I</scope><scope>8AF</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BENPR</scope><scope>BTHHO</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>M0S</scope><scope>M1P</scope><scope>M2P</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>Q9U</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>19870801</creationdate><title>Rapid technique of DNA-DNA in situ hybridisation on formalin fixed tissue sections using microwave irradiation</title><author>Coates, P J ; Hall, P A ; Butler, M G ; D'Ardenne, A J</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-b4215-6efcb9e11a42110841f6fb19a4a3ebe14dcbbd4254431ef8510762b7dcff54323</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1987</creationdate><topic>Analytical, structural and metabolic biochemistry</topic><topic>Biological and medical sciences</topic><topic>Cytomegalovirus - isolation & purification</topic><topic>Cytomegalovirus Infections - diagnosis</topic><topic>Cytomegalovirus Infections - microbiology</topic><topic>Dna, deoxyribonucleoproteins</topic><topic>DNA, Viral - analysis</topic><topic>Fundamental and applied biological sciences. 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| ispartof | Journal of clinical pathology, 1987-08, Vol.40 (8), p.865-869 |
| issn | 0021-9746 1472-4146 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_1141126 |
| source | MEDLINE; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central; Alma/SFX Local Collection |
| subjects | Analytical, structural and metabolic biochemistry Biological and medical sciences Cytomegalovirus - isolation & purification Cytomegalovirus Infections - diagnosis Cytomegalovirus Infections - microbiology Dna, deoxyribonucleoproteins DNA, Viral - analysis Fundamental and applied biological sciences. Psychology Humans Microwaves Nucleic Acid Denaturation Nucleic Acid Hybridization Nucleic acids Temperature |
| title | Rapid technique of DNA-DNA in situ hybridisation on formalin fixed tissue sections using microwave irradiation |
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