Probe binding to host proteins: A cause for false positive signals in viroid detection by tissue hybridization
Molecular hybridization assay, especially involving the use of tissues directly, has been developed as a rapid, simple and important technique for plant pathogen detection and/or gene expression analysis on a large scale. In theory, this method relies on the specific binding of a labeled probe to a...
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Veröffentlicht in: | Virus research 2009-10, Vol.145 (1), p.26-30 |
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description | Molecular hybridization assay, especially involving the use of tissues directly, has been developed as a rapid, simple and important technique for plant pathogen detection and/or gene expression analysis on a large scale. In theory, this method relies on the specific binding of a labeled probe to a target nucleotide sequence. However, occasional false positive reactions can pose a problem in its application and the cause is often not well understood. Here, we show that in tissue-printing hybridization to detect
Peach latent mosaic viroid (PLMVd) strong signals could arise by interactions between the viroid probe and plant proteins. Such probe–protein interactions made it difficult to show significant correlations between viroid infection and the level of hybridization signals. These results challenge the traditional view that proteins may hamper PCR reactions but have no influence on molecular hybridization. They further demonstrate that such probe–protein interactions in a plant could compromise the quality of molecular hybridization assays for viroid detection. Our results uncovered an important source of false positive reactions in tissue-printing hybridization and suggest that specificity can be improved by removing proteins. |
doi_str_mv | 10.1016/j.virusres.2009.06.011 |
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Peach latent mosaic viroid (PLMVd) strong signals could arise by interactions between the viroid probe and plant proteins. Such probe–protein interactions made it difficult to show significant correlations between viroid infection and the level of hybridization signals. These results challenge the traditional view that proteins may hamper PCR reactions but have no influence on molecular hybridization. They further demonstrate that such probe–protein interactions in a plant could compromise the quality of molecular hybridization assays for viroid detection. Our results uncovered an important source of false positive reactions in tissue-printing hybridization and suggest that specificity can be improved by removing proteins.</description><identifier>ISSN: 0168-1702</identifier><identifier>EISSN: 1872-7492</identifier><identifier>DOI: 10.1016/j.virusres.2009.06.011</identifier><identifier>PMID: 19540886</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>Deoxyribonuclease I - metabolism ; DNA Probes - metabolism ; DNA, Viral - genetics ; DNA, Viral - metabolism ; Endopeptidase K - metabolism ; False positive reaction ; False Positive Reactions ; Molecular hybridization ; Nucleic Acid Hybridization ; Peach latent mosaic viroid ; Plant Leaves - metabolism ; Plant Proteins - metabolism ; Plant Viruses - genetics ; Plant Viruses - isolation & purification ; Prunus - metabolism ; Prunus - virology ; Reverse Transcriptase Polymerase Chain Reaction ; Ribonuclease, Pancreatic - metabolism ; Sensitivity and Specificity ; Tissue dot-blot hybridization ; Tissue-print hybridization ; Viroids - genetics ; Viroids - isolation & purification</subject><ispartof>Virus research, 2009-10, Vol.145 (1), p.26-30</ispartof><rights>2009 Elsevier B.V.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c397t-fc1166e0eb69d0e5d7da5e346af998a05f80ae34204ff88e6918ef184e92751c3</citedby><cites>FETCH-LOGICAL-c397t-fc1166e0eb69d0e5d7da5e346af998a05f80ae34204ff88e6918ef184e92751c3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.virusres.2009.06.011$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/19540886$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>WenXing, Xu</creatorcontrib><creatorcontrib>Ni, Hong</creatorcontrib><creatorcontrib>QiuTing, Jin</creatorcontrib><creatorcontrib>Farooq, Abu Bakr Umer</creatorcontrib><creatorcontrib>ZeQiong, Wang</creatorcontrib><creatorcontrib>YanSu, Song</creatorcontrib><creatorcontrib>ChengChun, Wu</creatorcontrib><creatorcontrib>LiPing, Wang</creatorcontrib><creatorcontrib>GuoPing, Wang</creatorcontrib><title>Probe binding to host proteins: A cause for false positive signals in viroid detection by tissue hybridization</title><title>Virus research</title><addtitle>Virus Res</addtitle><description>Molecular hybridization assay, especially involving the use of tissues directly, has been developed as a rapid, simple and important technique for plant pathogen detection and/or gene expression analysis on a large scale. In theory, this method relies on the specific binding of a labeled probe to a target nucleotide sequence. However, occasional false positive reactions can pose a problem in its application and the cause is often not well understood. Here, we show that in tissue-printing hybridization to detect
Peach latent mosaic viroid (PLMVd) strong signals could arise by interactions between the viroid probe and plant proteins. Such probe–protein interactions made it difficult to show significant correlations between viroid infection and the level of hybridization signals. These results challenge the traditional view that proteins may hamper PCR reactions but have no influence on molecular hybridization. They further demonstrate that such probe–protein interactions in a plant could compromise the quality of molecular hybridization assays for viroid detection. Our results uncovered an important source of false positive reactions in tissue-printing hybridization and suggest that specificity can be improved by removing proteins.</description><subject>Deoxyribonuclease I - metabolism</subject><subject>DNA Probes - metabolism</subject><subject>DNA, Viral - genetics</subject><subject>DNA, Viral - metabolism</subject><subject>Endopeptidase K - metabolism</subject><subject>False positive reaction</subject><subject>False Positive Reactions</subject><subject>Molecular hybridization</subject><subject>Nucleic Acid Hybridization</subject><subject>Peach latent mosaic viroid</subject><subject>Plant Leaves - metabolism</subject><subject>Plant Proteins - metabolism</subject><subject>Plant Viruses - genetics</subject><subject>Plant Viruses - isolation & purification</subject><subject>Prunus - metabolism</subject><subject>Prunus - virology</subject><subject>Reverse Transcriptase Polymerase Chain Reaction</subject><subject>Ribonuclease, Pancreatic - metabolism</subject><subject>Sensitivity and Specificity</subject><subject>Tissue dot-blot hybridization</subject><subject>Tissue-print hybridization</subject><subject>Viroids - genetics</subject><subject>Viroids - isolation & purification</subject><issn>0168-1702</issn><issn>1872-7492</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2009</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkU9vEzEQxS1ERUPhK1Q-cdtlvH-8NieqihakSnAoZ8u7HrcTJXawdyOFT49Dgjj2ZHv0m3nj9xi7FlALEPLjut5TWnLCXDcAugZZgxCv2EqooamGTjev2aqAqhIDNJfsbc5rAJDtIN-wS6H7DpSSKxZ-pDgiHyk4Ck98jvw55pnvUpyRQv7Eb_hkl4zcx8S93ZTbLmaaaY8801MoFU6Bl2UiOe5wxmmmGPh44DPlvCB_PoyJHP22x_o7dvF3yPvzecV-3n15vP1aPXy__3Z781BNrR7myk9CSImAo9QOsHeDsz22nbRea2Wh9wpseTfQea8USi0UeqE61M3Qi6m9Yh9Oc8tHfi2YZ7OlPOFmYwPGJRs5yCIE_YtgA4NWQrcFlCdwSjEX373ZJdradDACzDESszb_IjHHSAxIUyIpjddnhWXcovvfds6gAJ9PABZD9oTJ5IkwTOgoFTeNi_SSxh9nE6LX</recordid><startdate>20091001</startdate><enddate>20091001</enddate><creator>WenXing, Xu</creator><creator>Ni, Hong</creator><creator>QiuTing, Jin</creator><creator>Farooq, Abu Bakr Umer</creator><creator>ZeQiong, Wang</creator><creator>YanSu, Song</creator><creator>ChengChun, Wu</creator><creator>LiPing, Wang</creator><creator>GuoPing, Wang</creator><general>Elsevier B.V</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>7U9</scope><scope>H94</scope><scope>7X8</scope></search><sort><creationdate>20091001</creationdate><title>Probe binding to host proteins: A cause for false positive signals in viroid detection by tissue hybridization</title><author>WenXing, Xu ; Ni, Hong ; QiuTing, Jin ; Farooq, Abu Bakr Umer ; ZeQiong, Wang ; YanSu, Song ; ChengChun, Wu ; LiPing, Wang ; GuoPing, Wang</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c397t-fc1166e0eb69d0e5d7da5e346af998a05f80ae34204ff88e6918ef184e92751c3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2009</creationdate><topic>Deoxyribonuclease I - metabolism</topic><topic>DNA Probes - metabolism</topic><topic>DNA, Viral - genetics</topic><topic>DNA, Viral - metabolism</topic><topic>Endopeptidase K - metabolism</topic><topic>False positive reaction</topic><topic>False Positive Reactions</topic><topic>Molecular hybridization</topic><topic>Nucleic Acid Hybridization</topic><topic>Peach latent mosaic viroid</topic><topic>Plant Leaves - metabolism</topic><topic>Plant Proteins - metabolism</topic><topic>Plant Viruses - genetics</topic><topic>Plant Viruses - isolation & purification</topic><topic>Prunus - metabolism</topic><topic>Prunus - virology</topic><topic>Reverse Transcriptase Polymerase Chain Reaction</topic><topic>Ribonuclease, Pancreatic - metabolism</topic><topic>Sensitivity and Specificity</topic><topic>Tissue dot-blot hybridization</topic><topic>Tissue-print hybridization</topic><topic>Viroids - genetics</topic><topic>Viroids - isolation & purification</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>WenXing, Xu</creatorcontrib><creatorcontrib>Ni, Hong</creatorcontrib><creatorcontrib>QiuTing, Jin</creatorcontrib><creatorcontrib>Farooq, Abu Bakr Umer</creatorcontrib><creatorcontrib>ZeQiong, Wang</creatorcontrib><creatorcontrib>YanSu, Song</creatorcontrib><creatorcontrib>ChengChun, Wu</creatorcontrib><creatorcontrib>LiPing, Wang</creatorcontrib><creatorcontrib>GuoPing, Wang</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Virology and AIDS Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Virus research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>WenXing, Xu</au><au>Ni, Hong</au><au>QiuTing, Jin</au><au>Farooq, Abu Bakr Umer</au><au>ZeQiong, Wang</au><au>YanSu, Song</au><au>ChengChun, Wu</au><au>LiPing, Wang</au><au>GuoPing, Wang</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Probe binding to host proteins: A cause for false positive signals in viroid detection by tissue hybridization</atitle><jtitle>Virus research</jtitle><addtitle>Virus Res</addtitle><date>2009-10-01</date><risdate>2009</risdate><volume>145</volume><issue>1</issue><spage>26</spage><epage>30</epage><pages>26-30</pages><issn>0168-1702</issn><eissn>1872-7492</eissn><abstract>Molecular hybridization assay, especially involving the use of tissues directly, has been developed as a rapid, simple and important technique for plant pathogen detection and/or gene expression analysis on a large scale. In theory, this method relies on the specific binding of a labeled probe to a target nucleotide sequence. However, occasional false positive reactions can pose a problem in its application and the cause is often not well understood. Here, we show that in tissue-printing hybridization to detect
Peach latent mosaic viroid (PLMVd) strong signals could arise by interactions between the viroid probe and plant proteins. Such probe–protein interactions made it difficult to show significant correlations between viroid infection and the level of hybridization signals. These results challenge the traditional view that proteins may hamper PCR reactions but have no influence on molecular hybridization. They further demonstrate that such probe–protein interactions in a plant could compromise the quality of molecular hybridization assays for viroid detection. Our results uncovered an important source of false positive reactions in tissue-printing hybridization and suggest that specificity can be improved by removing proteins.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>19540886</pmid><doi>10.1016/j.virusres.2009.06.011</doi><tpages>5</tpages></addata></record> |
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subjects | Deoxyribonuclease I - metabolism DNA Probes - metabolism DNA, Viral - genetics DNA, Viral - metabolism Endopeptidase K - metabolism False positive reaction False Positive Reactions Molecular hybridization Nucleic Acid Hybridization Peach latent mosaic viroid Plant Leaves - metabolism Plant Proteins - metabolism Plant Viruses - genetics Plant Viruses - isolation & purification Prunus - metabolism Prunus - virology Reverse Transcriptase Polymerase Chain Reaction Ribonuclease, Pancreatic - metabolism Sensitivity and Specificity Tissue dot-blot hybridization Tissue-print hybridization Viroids - genetics Viroids - isolation & purification |
title | Probe binding to host proteins: A cause for false positive signals in viroid detection by tissue hybridization |
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