Establishment and Evaluation of Stable Cell Lines Inhibiting Foot-and-Mouth Disease Virus by RNA Interference
RNA interference (RNAi) has been proved to be a powerful tool for foot-and-mouth disease virus FMDV inhibition in vitro and in vivo. We established five stable baby hamster kidney 21 cell lines (BHK-21) containing five short hairpin RNAs (shRNAs) expression plasmids (p3D1shRNA, p3D2shRNA, p3D3shRNA,...
Gespeichert in:
| Veröffentlicht in: | BioMed research international 2014-01, Vol.2014 (2014), p.1-7 |
|---|---|
| Hauptverfasser: | , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 7 |
|---|---|
| container_issue | 2014 |
| container_start_page | 1 |
| container_title | BioMed research international |
| container_volume | 2014 |
| creator | Zhang, Jie Gao, Zong-liang Zhou, Jian-hua Liu, Yong-sheng Gu, Yuan-xing |
| description | RNA interference (RNAi) has been proved to be a powerful tool for foot-and-mouth disease virus FMDV inhibition in vitro and in vivo. We established five stable baby hamster kidney 21 cell lines (BHK-21) containing five short hairpin RNAs (shRNAs) expression plasmids (p3D1shRNA, p3D2shRNA, p3D3shRNA, p3D4shRNA, and p3D5shRNA) targeting 3D gene of FMDV. Immunofluorescent assay, virus titration, and real-time quantitative reverse transcription polymerase chain reaction (Q-RT-PCR) were conducted to detect the effect of shRNAs on FMDV replication. After challenged with FMDV of O/CHA/99, two cell lines (p3D1shRNA and p3D4shRNA) showed a significant reduction in the synthesis of viral protein and RNA, accompanied by a sharp decrease in viral yield, and the inhibition could last for at least thirty passages. We developed an efficient procedure for the establishment and evaluation of stable cell lines for anti-FMDV research based on RNAi technology, which can be a candidate method for anti-FMDV research. |
| doi_str_mv | 10.1155/2014/109428 |
| format | Article |
| fullrecord | <record><control><sourceid>gale_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_3934452</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><galeid>A427021816</galeid><sourcerecordid>A427021816</sourcerecordid><originalsourceid>FETCH-LOGICAL-c527t-7ba341adccac879330131422d08325a3033af94f951d30946c9809749aa8678c3</originalsourceid><addsrcrecordid>eNqN0stv1DAQB-AIgWhVeuIMssQFgUJtj_PwpdJq2UKlBSReV2viOBtXiV3spKj_PV6lLI8L5OJI_vyTZzxZ9pjRV4wVxRmnTJwxKgWv72XHHJjISybY_cM_wFF2GuMVTV_NSirLh9kRF2UNBcjjbNzECZvBxn40biLoWrK5wWHGyXpHfEc-7bcNWZthIFvrTCSXrreNnazbkQvvpzydyd_5eerJaxsNRkO-2jBH0tySj-9XiU8mdCYYp82j7EGHQzSnd-tJ9uVi83n9Nt9-eHO5Xm1zXfBqyqsGQTBstUZdVxKAslQM5y2tgRcIFAA7KTpZsBZS7aWWNZWVkIh1WdUaTrLzJfd6bkbT6lRawEFdBztiuFUerfpzx9le7fyNAglCFDwFPL8LCP7bbOKkRht16gE64-eoWAGi5hxE-R-Ucc7T5Viiz_6iV34OLnUiKVowJoDJX2qHg1HWdT5dUe9D1UrwinKW3jGpl4vSwccYTHeojlG1Hw21Hw21jEbST39vyMH-HIQEXiygt67F7_YfaU8WbBIxHR6wEFWy8ANH-8bR</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>1505114319</pqid></control><display><type>article</type><title>Establishment and Evaluation of Stable Cell Lines Inhibiting Foot-and-Mouth Disease Virus by RNA Interference</title><source>MEDLINE</source><source>Wiley Online Library</source><source>PubMed Central</source><source>Alma/SFX Local Collection</source><source>PubMed Central Open Access</source><creator>Zhang, Jie ; Gao, Zong-liang ; Zhou, Jian-hua ; Liu, Yong-sheng ; Gu, Yuan-xing</creator><contributor>Dang, Zhisheng</contributor><creatorcontrib>Zhang, Jie ; Gao, Zong-liang ; Zhou, Jian-hua ; Liu, Yong-sheng ; Gu, Yuan-xing ; Dang, Zhisheng</creatorcontrib><description>RNA interference (RNAi) has been proved to be a powerful tool for foot-and-mouth disease virus FMDV inhibition in vitro and in vivo. We established five stable baby hamster kidney 21 cell lines (BHK-21) containing five short hairpin RNAs (shRNAs) expression plasmids (p3D1shRNA, p3D2shRNA, p3D3shRNA, p3D4shRNA, and p3D5shRNA) targeting 3D gene of FMDV. Immunofluorescent assay, virus titration, and real-time quantitative reverse transcription polymerase chain reaction (Q-RT-PCR) were conducted to detect the effect of shRNAs on FMDV replication. After challenged with FMDV of O/CHA/99, two cell lines (p3D1shRNA and p3D4shRNA) showed a significant reduction in the synthesis of viral protein and RNA, accompanied by a sharp decrease in viral yield, and the inhibition could last for at least thirty passages. We developed an efficient procedure for the establishment and evaluation of stable cell lines for anti-FMDV research based on RNAi technology, which can be a candidate method for anti-FMDV research.</description><identifier>ISSN: 2314-6133</identifier><identifier>EISSN: 2314-6141</identifier><identifier>DOI: 10.1155/2014/109428</identifier><identifier>PMID: 24683539</identifier><language>eng</language><publisher>Cairo, Egypt: Hindawi Puplishing Corporation</publisher><subject>Animals ; Cell Culture Techniques - methods ; Cell Line - virology ; Cell lines ; Cloning ; Cricetinae ; Deoxyribonucleic acid ; DNA ; Fluorescent Antibody Technique ; Foot & mouth disease ; Foot-and-mouth disease virus ; Foot-and-Mouth Disease Virus - physiology ; Genes ; Genetic aspects ; Genetic Vectors - metabolism ; Genomes ; Health aspects ; Plasmids ; Polymerase Chain Reaction ; Real-Time Polymerase Chain Reaction ; RNA Interference ; RNA polymerase ; RNA-protein interactions ; Rodents ; Science ; Serial Passage ; Transfection</subject><ispartof>BioMed research international, 2014-01, Vol.2014 (2014), p.1-7</ispartof><rights>Copyright © 2014 Yuan-xing Gu et al.</rights><rights>COPYRIGHT 2014 John Wiley & Sons, Inc.</rights><rights>Copyright © 2014 Yuan-xing Gu et al. Yuan-xing Gu et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.</rights><rights>Copyright © 2014 Yuan-xing Gu et al. 2014</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c527t-7ba341adccac879330131422d08325a3033af94f951d30946c9809749aa8678c3</citedby><cites>FETCH-LOGICAL-c527t-7ba341adccac879330131422d08325a3033af94f951d30946c9809749aa8678c3</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/PMC3934452/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC3934452/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,881,27901,27902,53766,53768</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24683539$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Dang, Zhisheng</contributor><creatorcontrib>Zhang, Jie</creatorcontrib><creatorcontrib>Gao, Zong-liang</creatorcontrib><creatorcontrib>Zhou, Jian-hua</creatorcontrib><creatorcontrib>Liu, Yong-sheng</creatorcontrib><creatorcontrib>Gu, Yuan-xing</creatorcontrib><title>Establishment and Evaluation of Stable Cell Lines Inhibiting Foot-and-Mouth Disease Virus by RNA Interference</title><title>BioMed research international</title><addtitle>Biomed Res Int</addtitle><description>RNA interference (RNAi) has been proved to be a powerful tool for foot-and-mouth disease virus FMDV inhibition in vitro and in vivo. We established five stable baby hamster kidney 21 cell lines (BHK-21) containing five short hairpin RNAs (shRNAs) expression plasmids (p3D1shRNA, p3D2shRNA, p3D3shRNA, p3D4shRNA, and p3D5shRNA) targeting 3D gene of FMDV. Immunofluorescent assay, virus titration, and real-time quantitative reverse transcription polymerase chain reaction (Q-RT-PCR) were conducted to detect the effect of shRNAs on FMDV replication. After challenged with FMDV of O/CHA/99, two cell lines (p3D1shRNA and p3D4shRNA) showed a significant reduction in the synthesis of viral protein and RNA, accompanied by a sharp decrease in viral yield, and the inhibition could last for at least thirty passages. We developed an efficient procedure for the establishment and evaluation of stable cell lines for anti-FMDV research based on RNAi technology, which can be a candidate method for anti-FMDV research.</description><subject>Animals</subject><subject>Cell Culture Techniques - methods</subject><subject>Cell Line - virology</subject><subject>Cell lines</subject><subject>Cloning</subject><subject>Cricetinae</subject><subject>Deoxyribonucleic acid</subject><subject>DNA</subject><subject>Fluorescent Antibody Technique</subject><subject>Foot & mouth disease</subject><subject>Foot-and-mouth disease virus</subject><subject>Foot-and-Mouth Disease Virus - physiology</subject><subject>Genes</subject><subject>Genetic aspects</subject><subject>Genetic Vectors - metabolism</subject><subject>Genomes</subject><subject>Health aspects</subject><subject>Plasmids</subject><subject>Polymerase Chain Reaction</subject><subject>Real-Time Polymerase Chain Reaction</subject><subject>RNA Interference</subject><subject>RNA polymerase</subject><subject>RNA-protein interactions</subject><subject>Rodents</subject><subject>Science</subject><subject>Serial Passage</subject><subject>Transfection</subject><issn>2314-6133</issn><issn>2314-6141</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>RHX</sourceid><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><recordid>eNqN0stv1DAQB-AIgWhVeuIMssQFgUJtj_PwpdJq2UKlBSReV2viOBtXiV3spKj_PV6lLI8L5OJI_vyTZzxZ9pjRV4wVxRmnTJwxKgWv72XHHJjISybY_cM_wFF2GuMVTV_NSirLh9kRF2UNBcjjbNzECZvBxn40biLoWrK5wWHGyXpHfEc-7bcNWZthIFvrTCSXrreNnazbkQvvpzydyd_5eerJaxsNRkO-2jBH0tySj-9XiU8mdCYYp82j7EGHQzSnd-tJ9uVi83n9Nt9-eHO5Xm1zXfBqyqsGQTBstUZdVxKAslQM5y2tgRcIFAA7KTpZsBZS7aWWNZWVkIh1WdUaTrLzJfd6bkbT6lRawEFdBztiuFUerfpzx9le7fyNAglCFDwFPL8LCP7bbOKkRht16gE64-eoWAGi5hxE-R-Ucc7T5Viiz_6iV34OLnUiKVowJoDJX2qHg1HWdT5dUe9D1UrwinKW3jGpl4vSwccYTHeojlG1Hw21Hw21jEbST39vyMH-HIQEXiygt67F7_YfaU8WbBIxHR6wEFWy8ANH-8bR</recordid><startdate>20140101</startdate><enddate>20140101</enddate><creator>Zhang, Jie</creator><creator>Gao, Zong-liang</creator><creator>Zhou, Jian-hua</creator><creator>Liu, Yong-sheng</creator><creator>Gu, Yuan-xing</creator><general>Hindawi Puplishing Corporation</general><general>Hindawi Publishing Corporation</general><general>John Wiley & Sons, Inc</general><general>Hindawi Limited</general><scope>ADJCN</scope><scope>AHFXO</scope><scope>RHU</scope><scope>RHW</scope><scope>RHX</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>7QL</scope><scope>7QO</scope><scope>7T7</scope><scope>7TK</scope><scope>7U7</scope><scope>7U9</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>CWDGH</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>7X8</scope><scope>7TM</scope><scope>5PM</scope></search><sort><creationdate>20140101</creationdate><title>Establishment and Evaluation of Stable Cell Lines Inhibiting Foot-and-Mouth Disease Virus by RNA Interference</title><author>Zhang, Jie ; Gao, Zong-liang ; Zhou, Jian-hua ; Liu, Yong-sheng ; Gu, Yuan-xing</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c527t-7ba341adccac879330131422d08325a3033af94f951d30946c9809749aa8678c3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Animals</topic><topic>Cell Culture Techniques - methods</topic><topic>Cell Line - virology</topic><topic>Cell lines</topic><topic>Cloning</topic><topic>Cricetinae</topic><topic>Deoxyribonucleic acid</topic><topic>DNA</topic><topic>Fluorescent Antibody Technique</topic><topic>Foot & mouth disease</topic><topic>Foot-and-mouth disease virus</topic><topic>Foot-and-Mouth Disease Virus - physiology</topic><topic>Genes</topic><topic>Genetic aspects</topic><topic>Genetic Vectors - metabolism</topic><topic>Genomes</topic><topic>Health aspects</topic><topic>Plasmids</topic><topic>Polymerase Chain Reaction</topic><topic>Real-Time Polymerase Chain Reaction</topic><topic>RNA Interference</topic><topic>RNA polymerase</topic><topic>RNA-protein interactions</topic><topic>Rodents</topic><topic>Science</topic><topic>Serial Passage</topic><topic>Transfection</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Zhang, Jie</creatorcontrib><creatorcontrib>Gao, Zong-liang</creatorcontrib><creatorcontrib>Zhou, Jian-hua</creatorcontrib><creatorcontrib>Liu, Yong-sheng</creatorcontrib><creatorcontrib>Gu, Yuan-xing</creatorcontrib><collection>الدوريات العلمية والإحصائية - e-Marefa Academic and Statistical Periodicals</collection><collection>معرفة - المحتوى العربي الأكاديمي المتكامل - e-Marefa Academic Complete</collection><collection>Hindawi Publishing Complete</collection><collection>Hindawi Publishing Subscription Journals</collection><collection>Hindawi Publishing Open Access</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Neurosciences Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Health Medical collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni)</collection><collection>ProQuest Central UK/Ireland</collection><collection>Advanced Technologies & Aerospace Database (1962 - current)</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>ProQuest Central</collection><collection>Technology Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>Middle East & Africa Database</collection><collection>ProQuest Central</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Biological Sciences</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>PML(ProQuest Medical Library)</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>ProQuest advanced technologies & aerospace journals</collection><collection>ProQuest Advanced Technologies & Aerospace Collection</collection><collection>Biotechnology and BioEngineering Abstracts</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>MEDLINE - Academic</collection><collection>Nucleic Acids Abstracts</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>BioMed research international</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Zhang, Jie</au><au>Gao, Zong-liang</au><au>Zhou, Jian-hua</au><au>Liu, Yong-sheng</au><au>Gu, Yuan-xing</au><au>Dang, Zhisheng</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Establishment and Evaluation of Stable Cell Lines Inhibiting Foot-and-Mouth Disease Virus by RNA Interference</atitle><jtitle>BioMed research international</jtitle><addtitle>Biomed Res Int</addtitle><date>2014-01-01</date><risdate>2014</risdate><volume>2014</volume><issue>2014</issue><spage>1</spage><epage>7</epage><pages>1-7</pages><issn>2314-6133</issn><eissn>2314-6141</eissn><abstract>RNA interference (RNAi) has been proved to be a powerful tool for foot-and-mouth disease virus FMDV inhibition in vitro and in vivo. We established five stable baby hamster kidney 21 cell lines (BHK-21) containing five short hairpin RNAs (shRNAs) expression plasmids (p3D1shRNA, p3D2shRNA, p3D3shRNA, p3D4shRNA, and p3D5shRNA) targeting 3D gene of FMDV. Immunofluorescent assay, virus titration, and real-time quantitative reverse transcription polymerase chain reaction (Q-RT-PCR) were conducted to detect the effect of shRNAs on FMDV replication. After challenged with FMDV of O/CHA/99, two cell lines (p3D1shRNA and p3D4shRNA) showed a significant reduction in the synthesis of viral protein and RNA, accompanied by a sharp decrease in viral yield, and the inhibition could last for at least thirty passages. We developed an efficient procedure for the establishment and evaluation of stable cell lines for anti-FMDV research based on RNAi technology, which can be a candidate method for anti-FMDV research.</abstract><cop>Cairo, Egypt</cop><pub>Hindawi Puplishing Corporation</pub><pmid>24683539</pmid><doi>10.1155/2014/109428</doi><tpages>7</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 2314-6133 |
| ispartof | BioMed research international, 2014-01, Vol.2014 (2014), p.1-7 |
| issn | 2314-6133 2314-6141 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_3934452 |
| source | MEDLINE; Wiley Online Library; PubMed Central; Alma/SFX Local Collection; PubMed Central Open Access |
| subjects | Animals Cell Culture Techniques - methods Cell Line - virology Cell lines Cloning Cricetinae Deoxyribonucleic acid DNA Fluorescent Antibody Technique Foot & mouth disease Foot-and-mouth disease virus Foot-and-Mouth Disease Virus - physiology Genes Genetic aspects Genetic Vectors - metabolism Genomes Health aspects Plasmids Polymerase Chain Reaction Real-Time Polymerase Chain Reaction RNA Interference RNA polymerase RNA-protein interactions Rodents Science Serial Passage Transfection |
| title | Establishment and Evaluation of Stable Cell Lines Inhibiting Foot-and-Mouth Disease Virus by RNA Interference |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2025-02-11T18%3A50%3A48IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-gale_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Establishment%20and%20Evaluation%20of%20Stable%20Cell%20Lines%20Inhibiting%20Foot-and-Mouth%20Disease%20Virus%20by%20RNA%20Interference&rft.jtitle=BioMed%20research%20international&rft.au=Zhang,%20Jie&rft.date=2014-01-01&rft.volume=2014&rft.issue=2014&rft.spage=1&rft.epage=7&rft.pages=1-7&rft.issn=2314-6133&rft.eissn=2314-6141&rft_id=info:doi/10.1155/2014/109428&rft_dat=%3Cgale_pubme%3EA427021816%3C/gale_pubme%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=1505114319&rft_id=info:pmid/24683539&rft_galeid=A427021816&rfr_iscdi=true |