Multiplex PCR detection of slowly-evolving trypanosomatids and neogregarines in bumblebees using broad-range primers
The aims of this study were to design universal markers for different protozoan parasites of Bombus spp. based on the phylogenetic position of two important bumblebee parasites Crithidia bombi and Apicystis bombi. Standard PCR and extraction techniques were used to amplify and sequence 18S rDNA. Phy...
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| Veröffentlicht in: | Journal of applied microbiology 2010-07, Vol.109 (1), p.107-115 |
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| description | The aims of this study were to design universal markers for different protozoan parasites of Bombus spp. based on the phylogenetic position of two important bumblebee parasites Crithidia bombi and Apicystis bombi. Standard PCR and extraction techniques were used to amplify and sequence 18S rDNA. Phylogenetic analysis of the rDNA was performed in order to predict the parasite range of the primers. Crithidia bombi phylogenetically clusters with the trypanosomatids with slowly-evolving SSU-rRNA sequences (SE), while A. bombi is the closest sister group of Mattesia. A multiplex was designed containing an internal control and two broad-range primer pairs, detecting C. bombi and other SE trypanosomatids and also A. bombi and other neogregarines. Sequence data generated will further improve the current systematics of insect trypanosomatids and gregarines that remain troublesome. Broad-range markers for bumblebee parasites are necessary tools enabling the screening of commercially imported colonies and thus controlling their worldwide distribution and to discover related emerging parasites. |
| doi_str_mv | 10.1111/j.1365-2672.2009.04635.x |
| format | Article |
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Standard PCR and extraction techniques were used to amplify and sequence 18S rDNA. Phylogenetic analysis of the rDNA was performed in order to predict the parasite range of the primers. Crithidia bombi phylogenetically clusters with the trypanosomatids with slowly-evolving SSU-rRNA sequences (SE), while A. bombi is the closest sister group of Mattesia. A multiplex was designed containing an internal control and two broad-range primer pairs, detecting C. bombi and other SE trypanosomatids and also A. bombi and other neogregarines. Sequence data generated will further improve the current systematics of insect trypanosomatids and gregarines that remain troublesome. Broad-range markers for bumblebee parasites are necessary tools enabling the screening of commercially imported colonies and thus controlling their worldwide distribution and to discover related emerging parasites.</description><identifier>ISSN: 1364-5072</identifier><identifier>EISSN: 1365-2672</identifier><identifier>DOI: 10.1111/j.1365-2672.2009.04635.x</identifier><identifier>PMID: 20015206</identifier><language>eng</language><publisher>Oxford, UK: Oxford, UK : Blackwell Publishing Ltd</publisher><subject>18S rDNA ; Animals ; Apicomplexa - genetics ; Apicomplexa - isolation & purification ; Apicystis bombi ; Bees - parasitology ; Biological and medical sciences ; Bombus ; Bumblebees ; Crithidia - genetics ; Crithidia - isolation & purification ; Crithidia bombi ; DNA Primers - genetics ; DNA, Protozoan - genetics ; Evolution ; Fundamental and applied biological sciences. Psychology ; Insects ; Markers ; Microbiology ; Multiplexing ; Parasites ; Phylogenetics ; Phylogeny ; Polymerase Chain Reaction - methods ; Protozoa ; protozoa detection ; RNA, Ribosomal, 18S - genetics ; rRNA ; Sequence Analysis, DNA ; Systematics</subject><ispartof>Journal of applied microbiology, 2010-07, Vol.109 (1), p.107-115</ispartof><rights>2009 The Authors. Journal compilation © 2009 The Society for Applied Microbiology</rights><rights>2015 INIST-CNRS</rights><rights>2009 The Authors. Journal compilation © 2009 The Society for Applied Microbiology.</rights><rights>Copyright Wiley Subscription Services, Inc. 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Standard PCR and extraction techniques were used to amplify and sequence 18S rDNA. Phylogenetic analysis of the rDNA was performed in order to predict the parasite range of the primers. Crithidia bombi phylogenetically clusters with the trypanosomatids with slowly-evolving SSU-rRNA sequences (SE), while A. bombi is the closest sister group of Mattesia. A multiplex was designed containing an internal control and two broad-range primer pairs, detecting C. bombi and other SE trypanosomatids and also A. bombi and other neogregarines. Sequence data generated will further improve the current systematics of insect trypanosomatids and gregarines that remain troublesome. Broad-range markers for bumblebee parasites are necessary tools enabling the screening of commercially imported colonies and thus controlling their worldwide distribution and to discover related emerging parasites.</description><subject>18S rDNA</subject><subject>Animals</subject><subject>Apicomplexa - genetics</subject><subject>Apicomplexa - isolation & purification</subject><subject>Apicystis bombi</subject><subject>Bees - parasitology</subject><subject>Biological and medical sciences</subject><subject>Bombus</subject><subject>Bumblebees</subject><subject>Crithidia - genetics</subject><subject>Crithidia - isolation & purification</subject><subject>Crithidia bombi</subject><subject>DNA Primers - genetics</subject><subject>DNA, Protozoan - genetics</subject><subject>Evolution</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Insects</subject><subject>Markers</subject><subject>Microbiology</subject><subject>Multiplexing</subject><subject>Parasites</subject><subject>Phylogenetics</subject><subject>Phylogeny</subject><subject>Polymerase Chain Reaction - methods</subject><subject>Protozoa</subject><subject>protozoa detection</subject><subject>RNA, Ribosomal, 18S - genetics</subject><subject>rRNA</subject><subject>Sequence Analysis, DNA</subject><subject>Systematics</subject><issn>1364-5072</issn><issn>1365-2672</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2010</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkstu1DAUhiMEohd4BbCEEKsE3-MsWFQjrmoFArq2bOckysgTD3bSzrx9nc5QJDbgjY_t7_c5x7-LAhFckTzerivCpCiprGlFMW4qzCUT1e5Rcfpw8Pg-5qXANT0pzlJaY0wYFvJpcZI1RFAsT4vpavbTsPWwQ99W31ELE7hpCCMKHUo-3Pp9CTfB3wxjj6a435oxpLAx09AmZMYWjRD6CL2JwwgJDSOy88Z6sJBXc1pUNgbTltGMPaBtHDYQ07PiSWd8gufH-by4_vD-5-pTefn14-fVxWXpBKOitMxZzKQT3KkGK8FrUNQKkXeEI5YzwNRyLCVpJORA2BoTaF1DGoUbDOy8eHO4dxvDrxnSpDdDcuC9yWXPSddCESaUYv8mOZdUMq4y-eovch3mOOY2NF2q5oyLOlPqQLkYUorQ6aV1E_eaYL1YqNd6cUovTunFQn1vod5l6YtjgtluoH0Q_vYsA6-PgEnO-C4_rRvSH44qxRvcZO7dgbsdPOz_uwD95eJqibL-5UHfmaBNH3OO6x90-UNECdo0gt0BJ3G_9Q</recordid><startdate>201007</startdate><enddate>201007</enddate><creator>Meeus, I</creator><creator>de Graaf, D.C</creator><creator>Jans, K</creator><creator>Smagghe, G</creator><general>Oxford, UK : Blackwell Publishing Ltd</general><general>Blackwell Publishing Ltd</general><general>Blackwell</general><general>Oxford University Press</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>7QL</scope><scope>7QO</scope><scope>7T7</scope><scope>7TM</scope><scope>7U7</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>7SS</scope><scope>7X8</scope></search><sort><creationdate>201007</creationdate><title>Multiplex PCR detection of slowly-evolving trypanosomatids and neogregarines in bumblebees using broad-range primers</title><author>Meeus, I ; de Graaf, D.C ; Jans, K ; Smagghe, G</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5325-b3cb036c54c8908547e82b556c55c1b43e02b4066196eb405b701edc9198090e3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>18S rDNA</topic><topic>Animals</topic><topic>Apicomplexa - genetics</topic><topic>Apicomplexa - isolation & purification</topic><topic>Apicystis bombi</topic><topic>Bees - parasitology</topic><topic>Biological and medical sciences</topic><topic>Bombus</topic><topic>Bumblebees</topic><topic>Crithidia - genetics</topic><topic>Crithidia - isolation & purification</topic><topic>Crithidia bombi</topic><topic>DNA Primers - genetics</topic><topic>DNA, Protozoan - genetics</topic><topic>Evolution</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Insects</topic><topic>Markers</topic><topic>Microbiology</topic><topic>Multiplexing</topic><topic>Parasites</topic><topic>Phylogenetics</topic><topic>Phylogeny</topic><topic>Polymerase Chain Reaction - methods</topic><topic>Protozoa</topic><topic>protozoa detection</topic><topic>RNA, Ribosomal, 18S - genetics</topic><topic>rRNA</topic><topic>Sequence Analysis, DNA</topic><topic>Systematics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Meeus, I</creatorcontrib><creatorcontrib>de Graaf, D.C</creatorcontrib><creatorcontrib>Jans, K</creatorcontrib><creatorcontrib>Smagghe, G</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>Bacteriology Abstracts (Microbiology B)</collection><collection>Biotechnology Research Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Nucleic Acids Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>MEDLINE - Academic</collection><jtitle>Journal of applied microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Meeus, I</au><au>de Graaf, D.C</au><au>Jans, K</au><au>Smagghe, G</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Multiplex PCR detection of slowly-evolving trypanosomatids and neogregarines in bumblebees using broad-range primers</atitle><jtitle>Journal of applied microbiology</jtitle><addtitle>J Appl Microbiol</addtitle><date>2010-07</date><risdate>2010</risdate><volume>109</volume><issue>1</issue><spage>107</spage><epage>115</epage><pages>107-115</pages><issn>1364-5072</issn><eissn>1365-2672</eissn><abstract>The aims of this study were to design universal markers for different protozoan parasites of Bombus spp. based on the phylogenetic position of two important bumblebee parasites Crithidia bombi and Apicystis bombi. Standard PCR and extraction techniques were used to amplify and sequence 18S rDNA. Phylogenetic analysis of the rDNA was performed in order to predict the parasite range of the primers. Crithidia bombi phylogenetically clusters with the trypanosomatids with slowly-evolving SSU-rRNA sequences (SE), while A. bombi is the closest sister group of Mattesia. A multiplex was designed containing an internal control and two broad-range primer pairs, detecting C. bombi and other SE trypanosomatids and also A. bombi and other neogregarines. Sequence data generated will further improve the current systematics of insect trypanosomatids and gregarines that remain troublesome. 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| source | Oxford University Press Journals All Titles (1996-Current); MEDLINE; Wiley Online Library Journals Frontfile Complete |
| subjects | 18S rDNA Animals Apicomplexa - genetics Apicomplexa - isolation & purification Apicystis bombi Bees - parasitology Biological and medical sciences Bombus Bumblebees Crithidia - genetics Crithidia - isolation & purification Crithidia bombi DNA Primers - genetics DNA, Protozoan - genetics Evolution Fundamental and applied biological sciences. Psychology Insects Markers Microbiology Multiplexing Parasites Phylogenetics Phylogeny Polymerase Chain Reaction - methods Protozoa protozoa detection RNA, Ribosomal, 18S - genetics rRNA Sequence Analysis, DNA Systematics |
| title | Multiplex PCR detection of slowly-evolving trypanosomatids and neogregarines in bumblebees using broad-range primers |
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