Detection of hepatitis A virus in shellfish (Mytilus galloprovincialis) with RT-PCR
A PCR assay for the detection of hepatitis A virus (HAV) in shellfish is described. The procedure involves the concentration of viral particles with the use of polyethylene glycol (PEG), followed by viral RNA extraction and purification with oligo(dT) cellulose. Reverse transcriptase-PCR detection w...
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| Veröffentlicht in: | Journal of food protection 2003-09, Vol.66 (9), p.1681-1685 |
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| container_title | Journal of food protection |
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| creator | Pinto, A. di Forte, V.T Tantillo, G.M Terio, V Buonavoglia, C |
| description | A PCR assay for the detection of hepatitis A virus (HAV) in shellfish is described. The procedure involves the concentration of viral particles with the use of polyethylene glycol (PEG), followed by viral RNA extraction and purification with oligo(dT) cellulose. Reverse transcriptase-PCR detection was accomplished in a single step with the use of primers specific for the VP3-VP1 region of the genome. The procedure detected one 50% tissue culture infective dose (0.6 PFU) per 25 g of shellfish homogenate. Heminested PCR was then carried out to verify the specificity of the PCR products. The method was used to detect HAV in shellfish samples from EU categories B and C and to evaluate the quality of shellfish in routine monitoring for HAV in view of the relevant public health implications of this foodborne disease. |
| doi_str_mv | 10.4315/0362-028X-66.9.1681 |
| format | Article |
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The procedure involves the concentration of viral particles with the use of polyethylene glycol (PEG), followed by viral RNA extraction and purification with oligo(dT) cellulose. Reverse transcriptase-PCR detection was accomplished in a single step with the use of primers specific for the VP3-VP1 region of the genome. The procedure detected one 50% tissue culture infective dose (0.6 PFU) per 25 g of shellfish homogenate. Heminested PCR was then carried out to verify the specificity of the PCR products. The method was used to detect HAV in shellfish samples from EU categories B and C and to evaluate the quality of shellfish in routine monitoring for HAV in view of the relevant public health implications of this foodborne disease.</description><identifier>ISSN: 0362-028X</identifier><identifier>EISSN: 1944-9097</identifier><identifier>DOI: 10.4315/0362-028X-66.9.1681</identifier><identifier>PMID: 14503725</identifier><identifier>CODEN: JFPRDR</identifier><language>eng</language><publisher>Des Moines, IA: International Association of Milk, Food and Environmental Sanitarians</publisher><subject>Animals ; assays ; Biological and medical sciences ; Bivalvia - microbiology ; Consumer Product Safety ; detection ; Fish and seafood industries ; food contamination ; Food industries ; Food microbiology ; food pathogens ; food safety ; Fundamental and applied biological sciences. Psychology ; Hepatitis A virus ; Hepatitis A virus - genetics ; Hepatitis A virus - isolation & purification ; mussels ; Mytilus galloprovincialis ; polymerase chain reaction ; real-time polymerase chain reaction ; Reverse Transcriptase Polymerase Chain Reaction - methods ; RNA, Viral - isolation & purification ; Sensitivity and Specificity ; shellfish ; Shellfish - microbiology ; Time Factors</subject><ispartof>Journal of food protection, 2003-09, Vol.66 (9), p.1681-1685</ispartof><rights>2004 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c431t-f4663c2012a430d767be6eb835dc514707e546e4b50279e5bc474d4db93e47fd3</citedby><cites>FETCH-LOGICAL-c431t-f4663c2012a430d767be6eb835dc514707e546e4b50279e5bc474d4db93e47fd3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>315,781,785,27929,27930</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=15125706$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/14503725$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Pinto, A. di</creatorcontrib><creatorcontrib>Forte, V.T</creatorcontrib><creatorcontrib>Tantillo, G.M</creatorcontrib><creatorcontrib>Terio, V</creatorcontrib><creatorcontrib>Buonavoglia, C</creatorcontrib><title>Detection of hepatitis A virus in shellfish (Mytilus galloprovincialis) with RT-PCR</title><title>Journal of food protection</title><addtitle>J Food Prot</addtitle><description>A PCR assay for the detection of hepatitis A virus (HAV) in shellfish is described. The procedure involves the concentration of viral particles with the use of polyethylene glycol (PEG), followed by viral RNA extraction and purification with oligo(dT) cellulose. Reverse transcriptase-PCR detection was accomplished in a single step with the use of primers specific for the VP3-VP1 region of the genome. The procedure detected one 50% tissue culture infective dose (0.6 PFU) per 25 g of shellfish homogenate. Heminested PCR was then carried out to verify the specificity of the PCR products. The method was used to detect HAV in shellfish samples from EU categories B and C and to evaluate the quality of shellfish in routine monitoring for HAV in view of the relevant public health implications of this foodborne disease.</description><subject>Animals</subject><subject>assays</subject><subject>Biological and medical sciences</subject><subject>Bivalvia - microbiology</subject><subject>Consumer Product Safety</subject><subject>detection</subject><subject>Fish and seafood industries</subject><subject>food contamination</subject><subject>Food industries</subject><subject>Food microbiology</subject><subject>food pathogens</subject><subject>food safety</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Hepatitis A virus</subject><subject>Hepatitis A virus - genetics</subject><subject>Hepatitis A virus - isolation & purification</subject><subject>mussels</subject><subject>Mytilus galloprovincialis</subject><subject>polymerase chain reaction</subject><subject>real-time polymerase chain reaction</subject><subject>Reverse Transcriptase Polymerase Chain Reaction - methods</subject><subject>RNA, Viral - isolation & purification</subject><subject>Sensitivity and Specificity</subject><subject>shellfish</subject><subject>Shellfish - microbiology</subject><subject>Time Factors</subject><issn>0362-028X</issn><issn>1944-9097</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2003</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkEtLAzEUhYMoWh-_QNBsFF1MvXlPlqU-oaJUBXchk8nYyLRTJ1PFf29Kiy5dXbh853D4EDok0OeMiAtgkmZA89dMyr7uE5mTDdQjmvNMg1abqPdL7KDdGN8BgGoqt9EO4QKYoqKHni59510XmhluKjzxc9uFLkQ8wJ-hXUQcZjhOfF1XIU7w2f13F-r0fbN13czb5jPMXLB1iOf4K3QTPH7OHofjfbRV2Tr6g_XdQy_XV8_D22z0cHM3HIwyl_Z3WcWlZI4CoZYzKJVUhZe-yJkonSBcgfKCS88LAVRpLwrHFS95WWjmuapKtodOV71pycfCx85MQ3RprJ35ZhGNEgqAU_UvSDRlQhOdQLYCXdvE2PrKzNswte23IWCW0s1SqVkqNVIabZbSU-poXb8opr78y6wtJ-BkDdjobF21NmmLf5wgNE2ViTtecZVtjH1rE_PylAQxIKDzXCj2AxnWkfk</recordid><startdate>20030901</startdate><enddate>20030901</enddate><creator>Pinto, A. di</creator><creator>Forte, V.T</creator><creator>Tantillo, G.M</creator><creator>Terio, V</creator><creator>Buonavoglia, C</creator><general>International Association of Milk, Food and Environmental Sanitarians</general><scope>FBQ</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>7T7</scope><scope>8FD</scope><scope>C1K</scope><scope>F1W</scope><scope>FR3</scope><scope>H97</scope><scope>L.G</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>20030901</creationdate><title>Detection of hepatitis A virus in shellfish (Mytilus galloprovincialis) with RT-PCR</title><author>Pinto, A. di ; Forte, V.T ; Tantillo, G.M ; Terio, V ; Buonavoglia, C</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c431t-f4663c2012a430d767be6eb835dc514707e546e4b50279e5bc474d4db93e47fd3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2003</creationdate><topic>Animals</topic><topic>assays</topic><topic>Biological and medical sciences</topic><topic>Bivalvia - microbiology</topic><topic>Consumer Product Safety</topic><topic>detection</topic><topic>Fish and seafood industries</topic><topic>food contamination</topic><topic>Food industries</topic><topic>Food microbiology</topic><topic>food pathogens</topic><topic>food safety</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Hepatitis A virus</topic><topic>Hepatitis A virus - genetics</topic><topic>Hepatitis A virus - isolation & purification</topic><topic>mussels</topic><topic>Mytilus galloprovincialis</topic><topic>polymerase chain reaction</topic><topic>real-time polymerase chain reaction</topic><topic>Reverse Transcriptase Polymerase Chain Reaction - methods</topic><topic>RNA, Viral - isolation & purification</topic><topic>Sensitivity and Specificity</topic><topic>shellfish</topic><topic>Shellfish - microbiology</topic><topic>Time Factors</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Pinto, A. di</creatorcontrib><creatorcontrib>Forte, V.T</creatorcontrib><creatorcontrib>Tantillo, G.M</creatorcontrib><creatorcontrib>Terio, V</creatorcontrib><creatorcontrib>Buonavoglia, C</creatorcontrib><collection>AGRIS</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>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ASFA: Aquatic Sciences and Fisheries Abstracts</collection><collection>Engineering Research Database</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) 3: Aquatic Pollution & Environmental Quality</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Professional</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of food protection</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Pinto, A. di</au><au>Forte, V.T</au><au>Tantillo, G.M</au><au>Terio, V</au><au>Buonavoglia, C</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Detection of hepatitis A virus in shellfish (Mytilus galloprovincialis) with RT-PCR</atitle><jtitle>Journal of food protection</jtitle><addtitle>J Food Prot</addtitle><date>2003-09-01</date><risdate>2003</risdate><volume>66</volume><issue>9</issue><spage>1681</spage><epage>1685</epage><pages>1681-1685</pages><issn>0362-028X</issn><eissn>1944-9097</eissn><coden>JFPRDR</coden><abstract>A PCR assay for the detection of hepatitis A virus (HAV) in shellfish is described. The procedure involves the concentration of viral particles with the use of polyethylene glycol (PEG), followed by viral RNA extraction and purification with oligo(dT) cellulose. Reverse transcriptase-PCR detection was accomplished in a single step with the use of primers specific for the VP3-VP1 region of the genome. The procedure detected one 50% tissue culture infective dose (0.6 PFU) per 25 g of shellfish homogenate. Heminested PCR was then carried out to verify the specificity of the PCR products. The method was used to detect HAV in shellfish samples from EU categories B and C and to evaluate the quality of shellfish in routine monitoring for HAV in view of the relevant public health implications of this foodborne disease.</abstract><cop>Des Moines, IA</cop><pub>International Association of Milk, Food and Environmental Sanitarians</pub><pmid>14503725</pmid><doi>10.4315/0362-028X-66.9.1681</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Animals assays Biological and medical sciences Bivalvia - microbiology Consumer Product Safety detection Fish and seafood industries food contamination Food industries Food microbiology food pathogens food safety Fundamental and applied biological sciences. Psychology Hepatitis A virus Hepatitis A virus - genetics Hepatitis A virus - isolation & purification mussels Mytilus galloprovincialis polymerase chain reaction real-time polymerase chain reaction Reverse Transcriptase Polymerase Chain Reaction - methods RNA, Viral - isolation & purification Sensitivity and Specificity shellfish Shellfish - microbiology Time Factors |
| title | Detection of hepatitis A virus in shellfish (Mytilus galloprovincialis) with RT-PCR |
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