Genotyping Trichomonas vaginalis
A genotyping method has been developed to distinguish each Trichomonas vaginalis isolate and has provided the first genome mapping studies of this protist with an estimated 180 Mb genome. The technique was developed using high molecular weight DNA prepared from five laboratory isolates from Australi...
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| Veröffentlicht in: | International journal for parasitology 2006-06, Vol.36 (7), p.821-828 |
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| container_title | International journal for parasitology |
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| creator | Upcroft, Jacqueline A. Delgadillo-Correa, Maria G. Dunne, Rebecca L. Sturm, A. Willem Johnson, Patricia J. Upcroft, Peter |
| description | A genotyping method has been developed to distinguish each
Trichomonas vaginalis isolate and has provided the first genome mapping studies of this protist with an estimated 180
Mb genome. The technique was developed using high molecular weight DNA prepared from five laboratory isolates from Australia and USA and 20 clinical isolates from South Africa. Inhibition of the notorious
T. vaginalis endogenous nucleases by addition of potent inhibitors was essential to the success of this study. Chromosomal DNA larger than 2.2
Mb was macrorestricted to a minimum segment size of ∼50
kb, separated by pulsed field gel electrophoresis and hybridised with a variety of gene probes. Each isolate generated a unique pattern that was distinguished by each of the probes. Four single copy gene probes (
fd,
hmp35,
ibp39 and
pfoD) were identified but probes which identified several bands (
pfoB and
α-scs) per isolate were most informative for genotyping. The pyruvate:ferredoxin oxidoreductase B gene probe identified two to seven copies of
pfoB (or its closely related homologue
pfoA) per genome in different isolates and is an obvious candidate probe to identify epidemiological linkage between infections by this genotyping method. Cleavage of the genomes into smaller fragments failed to distinguish isolates from diverse locations indicating the proximal regions of genes are conserved. |
| doi_str_mv | 10.1016/j.ijpara.2006.02.018 |
| format | Article |
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Trichomonas vaginalis isolate and has provided the first genome mapping studies of this protist with an estimated 180
Mb genome. The technique was developed using high molecular weight DNA prepared from five laboratory isolates from Australia and USA and 20 clinical isolates from South Africa. Inhibition of the notorious
T. vaginalis endogenous nucleases by addition of potent inhibitors was essential to the success of this study. Chromosomal DNA larger than 2.2
Mb was macrorestricted to a minimum segment size of ∼50
kb, separated by pulsed field gel electrophoresis and hybridised with a variety of gene probes. Each isolate generated a unique pattern that was distinguished by each of the probes. Four single copy gene probes (
fd,
hmp35,
ibp39 and
pfoD) were identified but probes which identified several bands (
pfoB and
α-scs) per isolate were most informative for genotyping. The pyruvate:ferredoxin oxidoreductase B gene probe identified two to seven copies of
pfoB (or its closely related homologue
pfoA) per genome in different isolates and is an obvious candidate probe to identify epidemiological linkage between infections by this genotyping method. Cleavage of the genomes into smaller fragments failed to distinguish isolates from diverse locations indicating the proximal regions of genes are conserved.</description><identifier>ISSN: 0020-7519</identifier><identifier>EISSN: 1879-0135</identifier><identifier>DOI: 10.1016/j.ijpara.2006.02.018</identifier><identifier>PMID: 16698025</identifier><identifier>CODEN: IJPYBT</identifier><language>eng</language><publisher>Oxford: Elsevier Ltd</publisher><subject>Animals ; Biological and medical sciences ; Chromosomal DNA ; DNA Probes ; DNA, Protozoan - genetics ; Electrophoresis, Gel, Pulsed-Field ; Endogenous nucleases ; Fundamental and applied biological sciences. Psychology ; Genome mapping ; Genome, Protozoan ; Genotype ; Humans ; Hybridisation ; Life cycle. Host-agent relationship. Pathogenesis ; Macrorestriction ; Protozoa ; Pulsed field gel electrophoresis ; Pyruvate Synthase - genetics ; Pyruvate:ferredoxin oxidoreductase ; Trichomonas vaginalis ; Trichomonas vaginalis - classification ; Trichomonas vaginalis - genetics ; Trichomonas vaginalis - isolation & purification</subject><ispartof>International journal for parasitology, 2006-06, Vol.36 (7), p.821-828</ispartof><rights>2006 Australian Society for Parasitology Inc</rights><rights>2006 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c421t-1f29fe2b87eeebfc1def9d386eb89a42d09b0c5aad3317119bb6213f6cf726743</citedby><cites>FETCH-LOGICAL-c421t-1f29fe2b87eeebfc1def9d386eb89a42d09b0c5aad3317119bb6213f6cf726743</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.ijpara.2006.02.018$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=17887013$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/16698025$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Upcroft, Jacqueline A.</creatorcontrib><creatorcontrib>Delgadillo-Correa, Maria G.</creatorcontrib><creatorcontrib>Dunne, Rebecca L.</creatorcontrib><creatorcontrib>Sturm, A. Willem</creatorcontrib><creatorcontrib>Johnson, Patricia J.</creatorcontrib><creatorcontrib>Upcroft, Peter</creatorcontrib><title>Genotyping Trichomonas vaginalis</title><title>International journal for parasitology</title><addtitle>Int J Parasitol</addtitle><description>A genotyping method has been developed to distinguish each
Trichomonas vaginalis isolate and has provided the first genome mapping studies of this protist with an estimated 180
Mb genome. The technique was developed using high molecular weight DNA prepared from five laboratory isolates from Australia and USA and 20 clinical isolates from South Africa. Inhibition of the notorious
T. vaginalis endogenous nucleases by addition of potent inhibitors was essential to the success of this study. Chromosomal DNA larger than 2.2
Mb was macrorestricted to a minimum segment size of ∼50
kb, separated by pulsed field gel electrophoresis and hybridised with a variety of gene probes. Each isolate generated a unique pattern that was distinguished by each of the probes. Four single copy gene probes (
fd,
hmp35,
ibp39 and
pfoD) were identified but probes which identified several bands (
pfoB and
α-scs) per isolate were most informative for genotyping. The pyruvate:ferredoxin oxidoreductase B gene probe identified two to seven copies of
pfoB (or its closely related homologue
pfoA) per genome in different isolates and is an obvious candidate probe to identify epidemiological linkage between infections by this genotyping method. Cleavage of the genomes into smaller fragments failed to distinguish isolates from diverse locations indicating the proximal regions of genes are conserved.</description><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Chromosomal DNA</subject><subject>DNA Probes</subject><subject>DNA, Protozoan - genetics</subject><subject>Electrophoresis, Gel, Pulsed-Field</subject><subject>Endogenous nucleases</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genome mapping</subject><subject>Genome, Protozoan</subject><subject>Genotype</subject><subject>Humans</subject><subject>Hybridisation</subject><subject>Life cycle. Host-agent relationship. Pathogenesis</subject><subject>Macrorestriction</subject><subject>Protozoa</subject><subject>Pulsed field gel electrophoresis</subject><subject>Pyruvate Synthase - genetics</subject><subject>Pyruvate:ferredoxin oxidoreductase</subject><subject>Trichomonas vaginalis</subject><subject>Trichomonas vaginalis - classification</subject><subject>Trichomonas vaginalis - genetics</subject><subject>Trichomonas vaginalis - isolation & purification</subject><issn>0020-7519</issn><issn>1879-0135</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2006</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqF0U1LAzEQgOEgiq3VfyDSi952ncnu5uMiiGgVBC_1HLLZiaZsd2vSCv33rrTQm55yeWYY3jB2iZAjoLhd5GGxstHmHEDkwHNAdcTGqKTOAIvqmI0BOGSyQj1iZyktALAqyvKUjVAIrYBXYzadUdevt6vQfUznMbjPftl3Nk2_7UfobBvSOTvxtk10sX8n7P3pcf7wnL2-zV4e7l8zV3JcZ-i59sRrJYmo9g4b8roplKBaaVvyBnQNrrK2KQqUiLquBcfCC-clF7IsJuxmt3cV-68NpbVZhuSobW1H_SYZoUCLSlX_QpQcSgVygOUOutinFMmbVQxLG7cGwfwmNAuzS2h-ExrgZkg4jF3t92_qJTWHoX2zAVzvgU3Otj7azoV0cFIpOXzA4O52joZs34GiSS5Q56gJkdzaNH34-5IfcfSQAg</recordid><startdate>20060601</startdate><enddate>20060601</enddate><creator>Upcroft, Jacqueline A.</creator><creator>Delgadillo-Correa, Maria G.</creator><creator>Dunne, Rebecca L.</creator><creator>Sturm, A. Willem</creator><creator>Johnson, Patricia J.</creator><creator>Upcroft, Peter</creator><general>Elsevier Ltd</general><general>Elsevier Science</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>8FD</scope><scope>FR3</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>20060601</creationdate><title>Genotyping Trichomonas vaginalis</title><author>Upcroft, Jacqueline A. ; Delgadillo-Correa, Maria G. ; Dunne, Rebecca L. ; Sturm, A. Willem ; Johnson, Patricia J. ; Upcroft, Peter</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c421t-1f29fe2b87eeebfc1def9d386eb89a42d09b0c5aad3317119bb6213f6cf726743</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2006</creationdate><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>Chromosomal DNA</topic><topic>DNA Probes</topic><topic>DNA, Protozoan - genetics</topic><topic>Electrophoresis, Gel, Pulsed-Field</topic><topic>Endogenous nucleases</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Genome mapping</topic><topic>Genome, Protozoan</topic><topic>Genotype</topic><topic>Humans</topic><topic>Hybridisation</topic><topic>Life cycle. Host-agent relationship. Pathogenesis</topic><topic>Macrorestriction</topic><topic>Protozoa</topic><topic>Pulsed field gel electrophoresis</topic><topic>Pyruvate Synthase - genetics</topic><topic>Pyruvate:ferredoxin oxidoreductase</topic><topic>Trichomonas vaginalis</topic><topic>Trichomonas vaginalis - classification</topic><topic>Trichomonas vaginalis - genetics</topic><topic>Trichomonas vaginalis - isolation & purification</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Upcroft, Jacqueline A.</creatorcontrib><creatorcontrib>Delgadillo-Correa, Maria G.</creatorcontrib><creatorcontrib>Dunne, Rebecca L.</creatorcontrib><creatorcontrib>Sturm, A. Willem</creatorcontrib><creatorcontrib>Johnson, Patricia J.</creatorcontrib><creatorcontrib>Upcroft, Peter</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>Technology Research Database</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>MEDLINE - Academic</collection><jtitle>International journal for parasitology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Upcroft, Jacqueline A.</au><au>Delgadillo-Correa, Maria G.</au><au>Dunne, Rebecca L.</au><au>Sturm, A. Willem</au><au>Johnson, Patricia J.</au><au>Upcroft, Peter</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Genotyping Trichomonas vaginalis</atitle><jtitle>International journal for parasitology</jtitle><addtitle>Int J Parasitol</addtitle><date>2006-06-01</date><risdate>2006</risdate><volume>36</volume><issue>7</issue><spage>821</spage><epage>828</epage><pages>821-828</pages><issn>0020-7519</issn><eissn>1879-0135</eissn><coden>IJPYBT</coden><abstract>A genotyping method has been developed to distinguish each
Trichomonas vaginalis isolate and has provided the first genome mapping studies of this protist with an estimated 180
Mb genome. The technique was developed using high molecular weight DNA prepared from five laboratory isolates from Australia and USA and 20 clinical isolates from South Africa. Inhibition of the notorious
T. vaginalis endogenous nucleases by addition of potent inhibitors was essential to the success of this study. Chromosomal DNA larger than 2.2
Mb was macrorestricted to a minimum segment size of ∼50
kb, separated by pulsed field gel electrophoresis and hybridised with a variety of gene probes. Each isolate generated a unique pattern that was distinguished by each of the probes. Four single copy gene probes (
fd,
hmp35,
ibp39 and
pfoD) were identified but probes which identified several bands (
pfoB and
α-scs) per isolate were most informative for genotyping. The pyruvate:ferredoxin oxidoreductase B gene probe identified two to seven copies of
pfoB (or its closely related homologue
pfoA) per genome in different isolates and is an obvious candidate probe to identify epidemiological linkage between infections by this genotyping method. Cleavage of the genomes into smaller fragments failed to distinguish isolates from diverse locations indicating the proximal regions of genes are conserved.</abstract><cop>Oxford</cop><pub>Elsevier Ltd</pub><pmid>16698025</pmid><doi>10.1016/j.ijpara.2006.02.018</doi><tpages>8</tpages></addata></record> |
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| ispartof | International journal for parasitology, 2006-06, Vol.36 (7), p.821-828 |
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| subjects | Animals Biological and medical sciences Chromosomal DNA DNA Probes DNA, Protozoan - genetics Electrophoresis, Gel, Pulsed-Field Endogenous nucleases Fundamental and applied biological sciences. Psychology Genome mapping Genome, Protozoan Genotype Humans Hybridisation Life cycle. Host-agent relationship. Pathogenesis Macrorestriction Protozoa Pulsed field gel electrophoresis Pyruvate Synthase - genetics Pyruvate:ferredoxin oxidoreductase Trichomonas vaginalis Trichomonas vaginalis - classification Trichomonas vaginalis - genetics Trichomonas vaginalis - isolation & purification |
| title | Genotyping Trichomonas vaginalis |
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