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Quantitative PCR demonstrates a positive correlation between a Rickettsia-like organism and severity of strawberry disease lesions in rainbow trout, Oncorhynchus mykiss (Walbaum)
Strawberry disease (SD) is an inflammatory skin disorder in rainbow trout, Oncorhynchus mykiss (Walbaum). The aetiology of SD is unknown although the 16S rDNA sequence of a Rickettsia‐like organism (RLO) has been associated with SD lesions using a nested PCR assay. In this study, we developed a Taqm...
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Published in: | Journal of fish diseases 2011-09, Vol.34 (9), p.701-709 |
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description | Strawberry disease (SD) is an inflammatory skin disorder in rainbow trout, Oncorhynchus mykiss (Walbaum). The aetiology of SD is unknown although the 16S rDNA sequence of a Rickettsia‐like organism (RLO) has been associated with SD lesions using a nested PCR assay. In this study, we developed a Taqman quantitative PCR assay (qPCR) that targeted the RLO 16S rDNA sequence to examine the distribution of RLO relative to lesion status. We compared 18 lesion samples from 13 fish representing high or low lesion severity as judged by gross examination. QPCR results showed that there was a higher number of RLO sequences in high severity lesions (mean of 12 068 copies) compared with fewer copies of RLO sequence in low severity lesions (mean of 3287 copies, P = 0.012). Grossly normal skin samples (n = 13) from SD‐affected fish were all negative by qPCR except two samples (121 and 139 copies). The qPCR assay described herein is a useful tool to investigate the role of RLO in SD in the absence of a culture system for RLO. Our results demonstrate a positive correlation between copy number and lesion severity consistent with the hypothesis that the RLO is the aetiologic agent of SD. |
doi_str_mv | 10.1111/j.1365-2761.2011.01285.x |
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The aetiology of SD is unknown although the 16S rDNA sequence of a Rickettsia‐like organism (RLO) has been associated with SD lesions using a nested PCR assay. In this study, we developed a Taqman quantitative PCR assay (qPCR) that targeted the RLO 16S rDNA sequence to examine the distribution of RLO relative to lesion status. We compared 18 lesion samples from 13 fish representing high or low lesion severity as judged by gross examination. QPCR results showed that there was a higher number of RLO sequences in high severity lesions (mean of 12 068 copies) compared with fewer copies of RLO sequence in low severity lesions (mean of 3287 copies, P = 0.012). Grossly normal skin samples (n = 13) from SD‐affected fish were all negative by qPCR except two samples (121 and 139 copies). The qPCR assay described herein is a useful tool to investigate the role of RLO in SD in the absence of a culture system for RLO. Our results demonstrate a positive correlation between copy number and lesion severity consistent with the hypothesis that the RLO is the aetiologic agent of SD.</description><identifier>ISSN: 0140-7775</identifier><identifier>EISSN: 1365-2761</identifier><identifier>DOI: 10.1111/j.1365-2761.2011.01285.x</identifier><identifier>PMID: 21838713</identifier><language>eng</language><publisher>Oxford, UK: Blackwell Publishing Ltd</publisher><subject>Animals ; Aquaculture ; DNA Primers - genetics ; Fish Diseases - microbiology ; Fish Diseases - pathology ; Fragaria ; Idaho ; Oncorhynchus mykiss ; qPCR ; rainbow trout ; Real-Time Polymerase Chain Reaction - veterinary ; Rickettsia - genetics ; Rickettsia Infections - genetics ; Rickettsia Infections - pathology ; Rickettsia Infections - veterinary ; Rickettsiaceae ; RNA, Ribosomal, 16S - genetics ; Skin Diseases - genetics ; Skin Diseases - pathology ; Skin Diseases - veterinary ; strawberry disease</subject><ispartof>Journal of fish diseases, 2011-09, Vol.34 (9), p.701-709</ispartof><rights>2011 Blackwell Publishing Ltd</rights><rights>2011 Blackwell Publishing Ltd.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c5155-cdf7beccc76c3185bcd817b8f78898c364335bc23ccc97793af3966b1cf544cc3</citedby><cites>FETCH-LOGICAL-c5155-cdf7beccc76c3185bcd817b8f78898c364335bc23ccc97793af3966b1cf544cc3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Fj.1365-2761.2011.01285.x$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Fj.1365-2761.2011.01285.x$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>315,786,790,27957,27958,50923,51032</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21838713$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Lloyd, S J</creatorcontrib><creatorcontrib>LaPatra, S E</creatorcontrib><creatorcontrib>Snekvik, K R</creatorcontrib><creatorcontrib>Cain, K D</creatorcontrib><creatorcontrib>Call, D R</creatorcontrib><title>Quantitative PCR demonstrates a positive correlation between a Rickettsia-like organism and severity of strawberry disease lesions in rainbow trout, Oncorhynchus mykiss (Walbaum)</title><title>Journal of fish diseases</title><addtitle>J Fish Dis</addtitle><description>Strawberry disease (SD) is an inflammatory skin disorder in rainbow trout, Oncorhynchus mykiss (Walbaum). The aetiology of SD is unknown although the 16S rDNA sequence of a Rickettsia‐like organism (RLO) has been associated with SD lesions using a nested PCR assay. In this study, we developed a Taqman quantitative PCR assay (qPCR) that targeted the RLO 16S rDNA sequence to examine the distribution of RLO relative to lesion status. We compared 18 lesion samples from 13 fish representing high or low lesion severity as judged by gross examination. QPCR results showed that there was a higher number of RLO sequences in high severity lesions (mean of 12 068 copies) compared with fewer copies of RLO sequence in low severity lesions (mean of 3287 copies, P = 0.012). Grossly normal skin samples (n = 13) from SD‐affected fish were all negative by qPCR except two samples (121 and 139 copies). The qPCR assay described herein is a useful tool to investigate the role of RLO in SD in the absence of a culture system for RLO. Our results demonstrate a positive correlation between copy number and lesion severity consistent with the hypothesis that the RLO is the aetiologic agent of SD.</description><subject>Animals</subject><subject>Aquaculture</subject><subject>DNA Primers - genetics</subject><subject>Fish Diseases - microbiology</subject><subject>Fish Diseases - pathology</subject><subject>Fragaria</subject><subject>Idaho</subject><subject>Oncorhynchus mykiss</subject><subject>qPCR</subject><subject>rainbow trout</subject><subject>Real-Time Polymerase Chain Reaction - veterinary</subject><subject>Rickettsia - genetics</subject><subject>Rickettsia Infections - genetics</subject><subject>Rickettsia Infections - pathology</subject><subject>Rickettsia Infections - veterinary</subject><subject>Rickettsiaceae</subject><subject>RNA, Ribosomal, 16S - genetics</subject><subject>Skin Diseases - genetics</subject><subject>Skin Diseases - pathology</subject><subject>Skin Diseases - veterinary</subject><subject>strawberry disease</subject><issn>0140-7775</issn><issn>1365-2761</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><recordid>eNqNkcFu1DAQhiMEotvCKyCLC1QiS7yOY-fQA1roFlQorUBFXCzHmVDvJvZiO93Na_GEON2yBy7gi62Z7_9Hnj9JEM6mOJ7XyykmBU1nrMDTWYbxNMMzTqfbB8lk33iYTDKcZyljjB4kh94vswwziovHycEMc8IZJpPk12UvTdBBBn0L6PP8CtXQWeODkwE8kmhtvb7rKesctJGzBlUQNgAmtq-0WkEIXsu01StA1v2QRvsOSVMjD7fgdBiQbdDouKnAuQHV2oP0gFrw0cwjbZCT2lR2g4KzfXiFLkycdjMYddN71A0r7T16eS3bSvbd8ZPkUSNbD0_v76Pk6-m7L_Oz9Pxi8X7-5jxVFFOaqrphFSilWKEI5rRSNces4g3jvOSKFDkhsTgjESkZK4lsSFkUFVYNzXOlyFHyYue7dvZnDz6ITnsFbSsN2N4LXpZx2QXP_03Gbedxwkg-_4tc2t6Z-I0RItEOZxHiO0g5672DRqyd7qQbBM7EmL9YijFmMcYsxvzFXf5iG6XP7v37qoN6L_wTeAROdsBGtzD8t7H4cPp2fEV9utNrH2C710u3EgUjjIrrTwvx8dsl_16cLQQlvwG0XdHz</recordid><startdate>201109</startdate><enddate>201109</enddate><creator>Lloyd, S J</creator><creator>LaPatra, S E</creator><creator>Snekvik, K R</creator><creator>Cain, K D</creator><creator>Call, D R</creator><general>Blackwell Publishing Ltd</general><scope>BSCLL</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>7QL</scope><scope>7TN</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>F1W</scope><scope>FR3</scope><scope>H94</scope><scope>H95</scope><scope>H98</scope><scope>H99</scope><scope>L.F</scope><scope>L.G</scope><scope>M7N</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>201109</creationdate><title>Quantitative PCR demonstrates a positive correlation between a Rickettsia-like organism and severity of strawberry disease lesions in rainbow trout, Oncorhynchus mykiss (Walbaum)</title><author>Lloyd, S J ; LaPatra, S E ; Snekvik, K R ; Cain, K D ; Call, D R</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5155-cdf7beccc76c3185bcd817b8f78898c364335bc23ccc97793af3966b1cf544cc3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>Animals</topic><topic>Aquaculture</topic><topic>DNA Primers - genetics</topic><topic>Fish Diseases - microbiology</topic><topic>Fish Diseases - pathology</topic><topic>Fragaria</topic><topic>Idaho</topic><topic>Oncorhynchus mykiss</topic><topic>qPCR</topic><topic>rainbow trout</topic><topic>Real-Time Polymerase Chain Reaction - veterinary</topic><topic>Rickettsia - genetics</topic><topic>Rickettsia Infections - genetics</topic><topic>Rickettsia Infections - pathology</topic><topic>Rickettsia Infections - veterinary</topic><topic>Rickettsiaceae</topic><topic>RNA, Ribosomal, 16S - genetics</topic><topic>Skin Diseases - genetics</topic><topic>Skin Diseases - pathology</topic><topic>Skin Diseases - veterinary</topic><topic>strawberry disease</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Lloyd, S J</creatorcontrib><creatorcontrib>LaPatra, S E</creatorcontrib><creatorcontrib>Snekvik, K R</creatorcontrib><creatorcontrib>Cain, K D</creatorcontrib><creatorcontrib>Call, D R</creatorcontrib><collection>Istex</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Oceanic Abstracts</collection><collection>Virology and AIDS Abstracts</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>AIDS and Cancer Research Abstracts</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) 1: Biological Sciences & Living Resources</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Aquaculture Abstracts</collection><collection>ASFA: Marine Biotechnology Abstracts</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Marine Biotechnology Abstracts</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Professional</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of fish diseases</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Lloyd, S J</au><au>LaPatra, S E</au><au>Snekvik, K R</au><au>Cain, K D</au><au>Call, D R</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Quantitative PCR demonstrates a positive correlation between a Rickettsia-like organism and severity of strawberry disease lesions in rainbow trout, Oncorhynchus mykiss (Walbaum)</atitle><jtitle>Journal of fish diseases</jtitle><addtitle>J Fish Dis</addtitle><date>2011-09</date><risdate>2011</risdate><volume>34</volume><issue>9</issue><spage>701</spage><epage>709</epage><pages>701-709</pages><issn>0140-7775</issn><eissn>1365-2761</eissn><notes>istex:512A2A322D96F36AC5233DE08AAFC24E424E0662</notes><notes>ArticleID:JFD1285</notes><notes>ark:/67375/WNG-MXQ8Z6HG-5</notes><notes>Present address: Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX 77555‐1070, USA.</notes><notes>ObjectType-Article-1</notes><notes>SourceType-Scholarly Journals-1</notes><notes>ObjectType-Feature-2</notes><notes>content type line 23</notes><abstract>Strawberry disease (SD) is an inflammatory skin disorder in rainbow trout, Oncorhynchus mykiss (Walbaum). The aetiology of SD is unknown although the 16S rDNA sequence of a Rickettsia‐like organism (RLO) has been associated with SD lesions using a nested PCR assay. In this study, we developed a Taqman quantitative PCR assay (qPCR) that targeted the RLO 16S rDNA sequence to examine the distribution of RLO relative to lesion status. We compared 18 lesion samples from 13 fish representing high or low lesion severity as judged by gross examination. QPCR results showed that there was a higher number of RLO sequences in high severity lesions (mean of 12 068 copies) compared with fewer copies of RLO sequence in low severity lesions (mean of 3287 copies, P = 0.012). Grossly normal skin samples (n = 13) from SD‐affected fish were all negative by qPCR except two samples (121 and 139 copies). The qPCR assay described herein is a useful tool to investigate the role of RLO in SD in the absence of a culture system for RLO. Our results demonstrate a positive correlation between copy number and lesion severity consistent with the hypothesis that the RLO is the aetiologic agent of SD.</abstract><cop>Oxford, UK</cop><pub>Blackwell Publishing Ltd</pub><pmid>21838713</pmid><doi>10.1111/j.1365-2761.2011.01285.x</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Animals Aquaculture DNA Primers - genetics Fish Diseases - microbiology Fish Diseases - pathology Fragaria Idaho Oncorhynchus mykiss qPCR rainbow trout Real-Time Polymerase Chain Reaction - veterinary Rickettsia - genetics Rickettsia Infections - genetics Rickettsia Infections - pathology Rickettsia Infections - veterinary Rickettsiaceae RNA, Ribosomal, 16S - genetics Skin Diseases - genetics Skin Diseases - pathology Skin Diseases - veterinary strawberry disease |
title | Quantitative PCR demonstrates a positive correlation between a Rickettsia-like organism and severity of strawberry disease lesions in rainbow trout, Oncorhynchus mykiss (Walbaum) |
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