Multiplex PCR assay for detection of four major bacterial pathogens causing rainbow trout disease
A multiplex PCR (mPCR) method was designed for the simultaneous detection of 4 major fish pathogens, Flavobacterium psychrophilum, Lactococcus garvieae, Pseudomonas aeruginosa, and P. putida. Each of the 4 pairs of oligonucleotide primers exclusively amplified the 16S rDNA gene of their targeted mic...
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| Veröffentlicht in: | Diseases of aquatic organisms 2011-02, Vol.93 (3), p.199-206 |
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| creator | ALTINOK, Ilhan |
| description | A multiplex PCR (mPCR) method was designed for the simultaneous detection of 4 major fish pathogens, Flavobacterium psychrophilum, Lactococcus garvieae, Pseudomonas aeruginosa, and P. putida. Each of the 4 pairs of oligonucleotide primers exclusively amplified the 16S rDNA gene of their targeted microorganism. The average detection limits for each organism amplified by mPCR were 2 colony-forming units (CFU) of F. psychrophilum, 3 CFU of L. garvieae, 3 CFU of P. aeruginosa, and 5 CFU of P. putida in mixed cultures. Multiplex PCR did not produce any nonspecific amplification products when tested against 28 related species of bacteria. High amounts of DNA from 1 bacterial species had a significant effect on the amplification sensitivity of the other bacterial species when these were present in lower concentrations in the multiplex reaction. The mPCR assay proved useful for the detection of the bacteria in naturally infected fish. The assay is a sensitive, specific, and reproducible diagnostic tool for the simultaneous detection of 4 pathogenic bacteria that cause disease in fish and offers a potentially useful alternative to the conventional culture-based method. |
| doi_str_mv | 10.3354/dao02300 |
| format | Article |
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Each of the 4 pairs of oligonucleotide primers exclusively amplified the 16S rDNA gene of their targeted microorganism. The average detection limits for each organism amplified by mPCR were 2 colony-forming units (CFU) of F. psychrophilum, 3 CFU of L. garvieae, 3 CFU of P. aeruginosa, and 5 CFU of P. putida in mixed cultures. Multiplex PCR did not produce any nonspecific amplification products when tested against 28 related species of bacteria. High amounts of DNA from 1 bacterial species had a significant effect on the amplification sensitivity of the other bacterial species when these were present in lower concentrations in the multiplex reaction. The mPCR assay proved useful for the detection of the bacteria in naturally infected fish. The assay is a sensitive, specific, and reproducible diagnostic tool for the simultaneous detection of 4 pathogenic bacteria that cause disease in fish and offers a potentially useful alternative to the conventional culture-based method.</description><identifier>ISSN: 0177-5103</identifier><identifier>EISSN: 1616-1580</identifier><identifier>DOI: 10.3354/dao02300</identifier><identifier>PMID: 21516972</identifier><identifier>CODEN: DAOREO</identifier><language>eng</language><publisher>Oldendorf: Inter-Research</publisher><subject>Animal aquaculture ; Animal productions ; Animals ; Bacteria ; Bacterial Infections - microbiology ; Bacterial Infections - veterinary ; Biological and medical sciences ; DNA, Bacterial ; Fish Diseases - diagnosis ; Fish Diseases - microbiology ; Flavobacterium psychrophilum ; Fundamental and applied biological sciences. Psychology ; Lactococcus garvieae ; Oncorhynchus mykiss ; Pisciculture ; Polymerase Chain Reaction - methods ; Polymerase Chain Reaction - veterinary ; Pseudomonas aeruginosa ; Vertebrate aquaculture</subject><ispartof>Diseases of aquatic organisms, 2011-02, Vol.93 (3), p.199-206</ispartof><rights>2015 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c378t-f8d1c003d8c6718025238d66ae62f0ab86abd19e3cc0a650ece6cf316ccc9a383</citedby><cites>FETCH-LOGICAL-c378t-f8d1c003d8c6718025238d66ae62f0ab86abd19e3cc0a650ece6cf316ccc9a383</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,3757,27923,27924</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=23942533$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21516972$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>ALTINOK, Ilhan</creatorcontrib><title>Multiplex PCR assay for detection of four major bacterial pathogens causing rainbow trout disease</title><title>Diseases of aquatic organisms</title><addtitle>Dis Aquat Organ</addtitle><description>A multiplex PCR (mPCR) method was designed for the simultaneous detection of 4 major fish pathogens, Flavobacterium psychrophilum, Lactococcus garvieae, Pseudomonas aeruginosa, and P. putida. Each of the 4 pairs of oligonucleotide primers exclusively amplified the 16S rDNA gene of their targeted microorganism. The average detection limits for each organism amplified by mPCR were 2 colony-forming units (CFU) of F. psychrophilum, 3 CFU of L. garvieae, 3 CFU of P. aeruginosa, and 5 CFU of P. putida in mixed cultures. Multiplex PCR did not produce any nonspecific amplification products when tested against 28 related species of bacteria. High amounts of DNA from 1 bacterial species had a significant effect on the amplification sensitivity of the other bacterial species when these were present in lower concentrations in the multiplex reaction. The mPCR assay proved useful for the detection of the bacteria in naturally infected fish. The assay is a sensitive, specific, and reproducible diagnostic tool for the simultaneous detection of 4 pathogenic bacteria that cause disease in fish and offers a potentially useful alternative to the conventional culture-based method.</description><subject>Animal aquaculture</subject><subject>Animal productions</subject><subject>Animals</subject><subject>Bacteria</subject><subject>Bacterial Infections - microbiology</subject><subject>Bacterial Infections - veterinary</subject><subject>Biological and medical sciences</subject><subject>DNA, Bacterial</subject><subject>Fish Diseases - diagnosis</subject><subject>Fish Diseases - microbiology</subject><subject>Flavobacterium psychrophilum</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Lactococcus garvieae</subject><subject>Oncorhynchus mykiss</subject><subject>Pisciculture</subject><subject>Polymerase Chain Reaction - methods</subject><subject>Polymerase Chain Reaction - veterinary</subject><subject>Pseudomonas aeruginosa</subject><subject>Vertebrate aquaculture</subject><issn>0177-5103</issn><issn>1616-1580</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqN0EFv1DAQBWALtaLbgsQvQL5UcAl4MhvHOaJVoZVagRCco8lkUlxl48V2RPffE9Rtufb0pKdP7_CUegPmA2K1_thTMCUa80KtwIItoHLmSK0M1HVRgcETdZrSnTFQNhW8VCclVGCbulwpupnH7Hej3Otvm--aUqK9HkLUvWTh7MOkw7AUc9Rbulv6jjhL9DTqHeVf4VampJnm5KdbHclPXfijcwxz1r1PQkleqeOBxiSvD3mmfn6--LG5LK6_frnafLouGGuXi8H1wMZg79jW4ExZleh6a0lsORjqnKWuh0aQ2ZCtjLBYHhAsMzeEDs_Uu4fdXQy_Z0m53frEMo40SZhT6-y6sQjwHIm1ta6BRb5_kBxDSlGGdhf9luK-BdP-e759fH6hbw-jc7eV_gk-Xr2A8wOgxDQOkSb26b_DZl1WiPgXwtSL7w</recordid><startdate>20110222</startdate><enddate>20110222</enddate><creator>ALTINOK, Ilhan</creator><general>Inter-Research</general><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>7X8</scope><scope>7QL</scope><scope>7T7</scope><scope>8FD</scope><scope>C1K</scope><scope>F1W</scope><scope>FR3</scope><scope>H95</scope><scope>H98</scope><scope>L.G</scope><scope>P64</scope></search><sort><creationdate>20110222</creationdate><title>Multiplex PCR assay for detection of four major bacterial pathogens causing rainbow trout disease</title><author>ALTINOK, Ilhan</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c378t-f8d1c003d8c6718025238d66ae62f0ab86abd19e3cc0a650ece6cf316ccc9a383</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>Animal aquaculture</topic><topic>Animal productions</topic><topic>Animals</topic><topic>Bacteria</topic><topic>Bacterial Infections - microbiology</topic><topic>Bacterial Infections - veterinary</topic><topic>Biological and medical sciences</topic><topic>DNA, Bacterial</topic><topic>Fish Diseases - diagnosis</topic><topic>Fish Diseases - microbiology</topic><topic>Flavobacterium psychrophilum</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Lactococcus garvieae</topic><topic>Oncorhynchus mykiss</topic><topic>Pisciculture</topic><topic>Polymerase Chain Reaction - methods</topic><topic>Polymerase Chain Reaction - veterinary</topic><topic>Pseudomonas aeruginosa</topic><topic>Vertebrate aquaculture</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>ALTINOK, Ilhan</creatorcontrib><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>MEDLINE - Academic</collection><collection>Bacteriology Abstracts (Microbiology B)</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) 1: Biological Sciences & Living Resources</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Aquaculture Abstracts</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Professional</collection><collection>Biotechnology and BioEngineering Abstracts</collection><jtitle>Diseases of aquatic organisms</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>ALTINOK, Ilhan</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Multiplex PCR assay for detection of four major bacterial pathogens causing rainbow trout disease</atitle><jtitle>Diseases of aquatic organisms</jtitle><addtitle>Dis Aquat Organ</addtitle><date>2011-02-22</date><risdate>2011</risdate><volume>93</volume><issue>3</issue><spage>199</spage><epage>206</epage><pages>199-206</pages><issn>0177-5103</issn><eissn>1616-1580</eissn><coden>DAOREO</coden><abstract>A multiplex PCR (mPCR) method was designed for the simultaneous detection of 4 major fish pathogens, Flavobacterium psychrophilum, Lactococcus garvieae, Pseudomonas aeruginosa, and P. putida. Each of the 4 pairs of oligonucleotide primers exclusively amplified the 16S rDNA gene of their targeted microorganism. The average detection limits for each organism amplified by mPCR were 2 colony-forming units (CFU) of F. psychrophilum, 3 CFU of L. garvieae, 3 CFU of P. aeruginosa, and 5 CFU of P. putida in mixed cultures. Multiplex PCR did not produce any nonspecific amplification products when tested against 28 related species of bacteria. High amounts of DNA from 1 bacterial species had a significant effect on the amplification sensitivity of the other bacterial species when these were present in lower concentrations in the multiplex reaction. The mPCR assay proved useful for the detection of the bacteria in naturally infected fish. 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| ispartof | Diseases of aquatic organisms, 2011-02, Vol.93 (3), p.199-206 |
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| source | MEDLINE; Inter-Research; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Alma/SFX Local Collection |
| subjects | Animal aquaculture Animal productions Animals Bacteria Bacterial Infections - microbiology Bacterial Infections - veterinary Biological and medical sciences DNA, Bacterial Fish Diseases - diagnosis Fish Diseases - microbiology Flavobacterium psychrophilum Fundamental and applied biological sciences. Psychology Lactococcus garvieae Oncorhynchus mykiss Pisciculture Polymerase Chain Reaction - methods Polymerase Chain Reaction - veterinary Pseudomonas aeruginosa Vertebrate aquaculture |
| title | Multiplex PCR assay for detection of four major bacterial pathogens causing rainbow trout disease |
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