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Use of an RT-PCR internal control to evaluate viral removal
We constructed an internal control based on the poliovirus genome and cloned it into a transcription plasmid to obtain a competitor RNA distinguishable by its size from the wild-type PCR product. A known number of copies was introduced into sewage samples before nucleic acid extraction in order to e...
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| Published in: | Water science and technology 1997-01, Vol.35 (11-12), p.461-465 |
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| Main Authors: | , , , , , |
| Format: | Article |
| Language: | English |
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| cited_by | cdi_FETCH-LOGICAL-c442t-f35dadd51da801c03bc6a8d488a17cd768b2c3090e9e8cb758cc940da2c08ae13 |
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| container_end_page | 465 |
| container_issue | 11-12 |
| container_start_page | 461 |
| container_title | Water science and technology |
| container_volume | 35 |
| creator | Le Guyader, F. Menard, D. Dubois, E. Haugarreau, L. Kopecka, H. Pommepuy, M. |
| description | We constructed an internal control based on the poliovirus genome and cloned it into a transcription plasmid to obtain a competitor RNA distinguishable by its size from the wild-type PCR product. A known number of copies was introduced into sewage samples before nucleic acid extraction in order to evaluate the reliability of the extraction step and amplification. The final gel was analysed with a densitometer and the number of viral RNA copies present in the sewage was calculated. In vitro experiments were carried Out with this internal control with peracetic acid and UV to estimate decontamination efficiency. A very high concentration of peracetic acid was required to eliminate viruses with a moderate UV dose to obtain a negative PCR. This study demonstrates that competitor RNA is useful both in controlling the different reaction steps (extraction and amplification) and inhibitor detection and in providing a quantitative estimation of contamination. |
| doi_str_mv | 10.1016/S0273-1223(97)00304-1 |
| format | article |
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A known number of copies was introduced into sewage samples before nucleic acid extraction in order to evaluate the reliability of the extraction step and amplification. The final gel was analysed with a densitometer and the number of viral RNA copies present in the sewage was calculated. In vitro experiments were carried Out with this internal control with peracetic acid and UV to estimate decontamination efficiency. A very high concentration of peracetic acid was required to eliminate viruses with a moderate UV dose to obtain a negative PCR. 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Psychology ; Gels ; General purification processes ; Genomes ; internal control ; Microbiology ; Nucleic acids ; Nucleotide sequence ; PCR ; Peracetic acid ; Plasmids ; Pollution ; Polymerase chain reaction ; Reliability analysis ; Removal ; Ribonucleic acid ; RNA ; Sewage ; Techniques used in virology ; Transcription ; Ultraviolet radiation ; Virology ; Viruses ; Wastewaters ; Water treatment and pollution</subject><ispartof>Water science and technology, 1997-01, Vol.35 (11-12), p.461-465</ispartof><rights>1997 International Association on Water Quality</rights><rights>1997 INIST-CNRS</rights><rights>Copyright IWA Publishing Jun 1997</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c442t-f35dadd51da801c03bc6a8d488a17cd768b2c3090e9e8cb758cc940da2c08ae13</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>309,310,314,777,781,786,787,27905,27906</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=2734657$$DView record in Pascal Francis$$Hfree_for_read</backlink></links><search><contributor>Grabow, WOK</contributor><contributor>Morris, R</contributor><contributor>Jofre, J (eds)</contributor><creatorcontrib>Le Guyader, F.</creatorcontrib><creatorcontrib>Menard, D.</creatorcontrib><creatorcontrib>Dubois, E.</creatorcontrib><creatorcontrib>Haugarreau, L.</creatorcontrib><creatorcontrib>Kopecka, H.</creatorcontrib><creatorcontrib>Pommepuy, M.</creatorcontrib><title>Use of an RT-PCR internal control to evaluate viral removal</title><title>Water science and technology</title><description>We constructed an internal control based on the poliovirus genome and cloned it into a transcription plasmid to obtain a competitor RNA distinguishable by its size from the wild-type PCR product. 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Psychology</topic><topic>Gels</topic><topic>General purification processes</topic><topic>Genomes</topic><topic>internal control</topic><topic>Microbiology</topic><topic>Nucleic acids</topic><topic>Nucleotide sequence</topic><topic>PCR</topic><topic>Peracetic acid</topic><topic>Plasmids</topic><topic>Pollution</topic><topic>Polymerase chain reaction</topic><topic>Reliability analysis</topic><topic>Removal</topic><topic>Ribonucleic acid</topic><topic>RNA</topic><topic>Sewage</topic><topic>Techniques used in virology</topic><topic>Transcription</topic><topic>Ultraviolet radiation</topic><topic>Virology</topic><topic>Viruses</topic><topic>Wastewaters</topic><topic>Water treatment and pollution</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Le Guyader, F.</creatorcontrib><creatorcontrib>Menard, D.</creatorcontrib><creatorcontrib>Dubois, E.</creatorcontrib><creatorcontrib>Haugarreau, L.</creatorcontrib><creatorcontrib>Kopecka, H.</creatorcontrib><creatorcontrib>Pommepuy, M.</creatorcontrib><collection>Pascal-Francis</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Aqualine</collection><collection>Water Resources Abstracts</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Technology Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>Materials Science & Engineering Collection</collection><collection>ProQuest Central (Alumni)</collection><collection>ProQuest One Sustainability</collection><collection>ProQuest Central</collection><collection>ProQuest Databases</collection><collection>Technology Collection</collection><collection>Natural Science Collection</collection><collection>Earth, Atmospheric & Aquatic Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central</collection><collection>ASFA: Aquatic Sciences and Fisheries Abstracts</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) 2: Ocean Technology, Policy & Non-Living Resources</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) 3: Aquatic Pollution & Environmental Quality</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Professional</collection><collection>ProQuest Engineering Collection</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Engineering Database</collection><collection>Earth, Atmospheric & Aquatic Science 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>Engineering Collection</collection><jtitle>Water science and technology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Le Guyader, F.</au><au>Menard, D.</au><au>Dubois, E.</au><au>Haugarreau, L.</au><au>Kopecka, H.</au><au>Pommepuy, M.</au><au>Grabow, WOK</au><au>Morris, R</au><au>Jofre, J (eds)</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Use of an RT-PCR internal control to evaluate viral removal</atitle><jtitle>Water science and technology</jtitle><date>1997-01-01</date><risdate>1997</risdate><volume>35</volume><issue>11-12</issue><spage>461</spage><epage>465</epage><pages>461-465</pages><issn>0273-1223</issn><eissn>1996-9732</eissn><isbn>9780080433004</isbn><isbn>0080433006</isbn><coden>WSTED4</coden><abstract>We constructed an internal control based on the poliovirus genome and cloned it into a transcription plasmid to obtain a competitor RNA distinguishable by its size from the wild-type PCR product. A known number of copies was introduced into sewage samples before nucleic acid extraction in order to evaluate the reliability of the extraction step and amplification. The final gel was analysed with a densitometer and the number of viral RNA copies present in the sewage was calculated. In vitro experiments were carried Out with this internal control with peracetic acid and UV to estimate decontamination efficiency. A very high concentration of peracetic acid was required to eliminate viruses with a moderate UV dose to obtain a negative PCR. This study demonstrates that competitor RNA is useful both in controlling the different reaction steps (extraction and amplification) and inhibitor detection and in providing a quantitative estimation of contamination.</abstract><cop>New York NY</cop><cop>Oxford</cop><pub>Elsevier Ltd</pub><doi>10.1016/S0273-1223(97)00304-1</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0273-1223 |
| ispartof | Water science and technology, 1997-01, Vol.35 (11-12), p.461-465 |
| issn | 0273-1223 1996-9732 |
| language | eng |
| recordid | cdi_proquest_miscellaneous_16463560 |
| source | Alma/SFX Local Collection |
| subjects | Amplification Applied sciences Biological and medical sciences competitive RT-PCR Contamination Control Decontamination Densitometers Detection DNA Enterovirus Exact sciences and technology Extraction Fundamental and applied biological sciences. Psychology Gels General purification processes Genomes internal control Microbiology Nucleic acids Nucleotide sequence PCR Peracetic acid Plasmids Pollution Polymerase chain reaction Reliability analysis Removal Ribonucleic acid RNA Sewage Techniques used in virology Transcription Ultraviolet radiation Virology Viruses Wastewaters Water treatment and pollution |
| title | Use of an RT-PCR internal control to evaluate viral removal |
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