Efficacy of magnetic capture in comparison with conventional DNA isolation in a survey of Toxoplasma gondii in wild house mice
Toxoplasma gondii is a zoonotic parasite with a world-wide distribution. House mice (Mus musculus) play an important role as a reservoir host in the parasite life cycle. However, their detection in mouse brain is limited because the host potentially harbours only a few tissue cysts. In order to impr...
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Veröffentlicht in: | European journal of protistology 2014-02, Vol.50 (1), p.11-15 |
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creator | Juránková, Jana Hůrková-Hofmannová, Lada Volf, Jiří Baláž, Vojtech Piálek, Jaroslav |
description | Toxoplasma gondii is a zoonotic parasite with a world-wide distribution. House mice (Mus musculus) play an important role as a reservoir host in the parasite life cycle. However, their detection in mouse brain is limited because the host potentially harbours only a few tissue cysts. In order to improve the diagnosis, we tested a novel protocol for T. gondii detection in mice and compared this technique to a standard PCR-based protocol using a commercial kit for DNA isolation. Efficacy of magnetic capture for isolation of T. gondii DNA from whole host brains was tested in brain samples of laboratory mice spiked with 1 up to 104 tachyzoites. Real-time PCR revealed that even 1–5 tachyzoites can be detected after magnetic capture. Also this method is suitable to quantify parasite numbers in mouse brains with more than 10 tachyzoite equivalents. To assess the two techniques in wild mice, we employed a dataset consisting of 243 individuals. The prevalence of T. gondii detected by magnetic capture and qPCR and by commercial isolation and PCR was 1.2% and 0%, respectively. The magnetic capture and quantitative PCR seems to be a highly sensitive and specific diagnostic method for both laboratory research and wild population surveys. |
doi_str_mv | 10.1016/j.ejop.2013.08.002 |
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House mice (Mus musculus) play an important role as a reservoir host in the parasite life cycle. However, their detection in mouse brain is limited because the host potentially harbours only a few tissue cysts. In order to improve the diagnosis, we tested a novel protocol for T. gondii detection in mice and compared this technique to a standard PCR-based protocol using a commercial kit for DNA isolation. Efficacy of magnetic capture for isolation of T. gondii DNA from whole host brains was tested in brain samples of laboratory mice spiked with 1 up to 104 tachyzoites. Real-time PCR revealed that even 1–5 tachyzoites can be detected after magnetic capture. Also this method is suitable to quantify parasite numbers in mouse brains with more than 10 tachyzoite equivalents. To assess the two techniques in wild mice, we employed a dataset consisting of 243 individuals. The prevalence of T. gondii detected by magnetic capture and qPCR and by commercial isolation and PCR was 1.2% and 0%, respectively. The magnetic capture and quantitative PCR seems to be a highly sensitive and specific diagnostic method for both laboratory research and wild population surveys.</description><identifier>ISSN: 0932-4739</identifier><identifier>EISSN: 1618-0429</identifier><identifier>DOI: 10.1016/j.ejop.2013.08.002</identifier><identifier>PMID: 24280460</identifier><language>eng</language><publisher>Munich: Elsevier GmbH</publisher><subject>Animal protozoal diseases ; Animals ; Biological and medical sciences ; DNA, Protozoan - analysis ; DNA, Protozoan - isolation & purification ; Female ; Fundamental and applied biological sciences. Psychology ; Genetic Techniques - standards ; House mice ; Infectious diseases ; Life cycle. Host-agent relationship. Pathogenesis ; Limit of Detection ; Magnetic capture ; Magnetic Phenomena ; Medical sciences ; Mice ; Mus musculus ; Parasitic diseases ; Polymerase Chain Reaction - standards ; Protozoa ; Protozoal diseases ; Quantitative PCR ; Toxoplasma - genetics ; Toxoplasma gondii ; Toxoplasmosis, Animal - diagnosis</subject><ispartof>European journal of protistology, 2014-02, Vol.50 (1), p.11-15</ispartof><rights>2013 Elsevier GmbH</rights><rights>2015 INIST-CNRS</rights><rights>Copyright © 2013 Elsevier GmbH. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c419t-90b14953c72abbb7bd244c6277e7607f69df00c9970b0ed55fc25ad7ce8437103</citedby><cites>FETCH-LOGICAL-c419t-90b14953c72abbb7bd244c6277e7607f69df00c9970b0ed55fc25ad7ce8437103</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0932473913000692$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27901,27902,65306</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=28150561$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24280460$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Juránková, Jana</creatorcontrib><creatorcontrib>Hůrková-Hofmannová, Lada</creatorcontrib><creatorcontrib>Volf, Jiří</creatorcontrib><creatorcontrib>Baláž, Vojtech</creatorcontrib><creatorcontrib>Piálek, Jaroslav</creatorcontrib><title>Efficacy of magnetic capture in comparison with conventional DNA isolation in a survey of Toxoplasma gondii in wild house mice</title><title>European journal of protistology</title><addtitle>Eur J Protistol</addtitle><description>Toxoplasma gondii is a zoonotic parasite with a world-wide distribution. House mice (Mus musculus) play an important role as a reservoir host in the parasite life cycle. However, their detection in mouse brain is limited because the host potentially harbours only a few tissue cysts. In order to improve the diagnosis, we tested a novel protocol for T. gondii detection in mice and compared this technique to a standard PCR-based protocol using a commercial kit for DNA isolation. Efficacy of magnetic capture for isolation of T. gondii DNA from whole host brains was tested in brain samples of laboratory mice spiked with 1 up to 104 tachyzoites. Real-time PCR revealed that even 1–5 tachyzoites can be detected after magnetic capture. Also this method is suitable to quantify parasite numbers in mouse brains with more than 10 tachyzoite equivalents. To assess the two techniques in wild mice, we employed a dataset consisting of 243 individuals. The prevalence of T. gondii detected by magnetic capture and qPCR and by commercial isolation and PCR was 1.2% and 0%, respectively. The magnetic capture and quantitative PCR seems to be a highly sensitive and specific diagnostic method for both laboratory research and wild population surveys.</description><subject>Animal protozoal diseases</subject><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>DNA, Protozoan - analysis</subject><subject>DNA, Protozoan - isolation & purification</subject><subject>Female</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Genetic Techniques - standards</subject><subject>House mice</subject><subject>Infectious diseases</subject><subject>Life cycle. Host-agent relationship. Pathogenesis</subject><subject>Limit of Detection</subject><subject>Magnetic capture</subject><subject>Magnetic Phenomena</subject><subject>Medical sciences</subject><subject>Mice</subject><subject>Mus musculus</subject><subject>Parasitic diseases</subject><subject>Polymerase Chain Reaction - standards</subject><subject>Protozoa</subject><subject>Protozoal diseases</subject><subject>Quantitative PCR</subject><subject>Toxoplasma - genetics</subject><subject>Toxoplasma gondii</subject><subject>Toxoplasmosis, Animal - diagnosis</subject><issn>0932-4739</issn><issn>1618-0429</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkU1v1DAQhi0EotvCH-CAfEHiknTsOHEscalKP5AquJSz5TiT1qvEDnaybS_8dhJ2gVs5WSM_8-rVPIS8Y5AzYNXpNsdtGHMOrMihzgH4C7JhFaszEFy9JBtQBc-ELNQROU5pCwCKVeVrcsQFr0FUsCE_L7rOWWOfaOjoYO48Ts5Sa8ZpjkidpzYMo4kuBU8f3HS_zH6HfnLBm55-_npGl6_erPNKG5rmuMPfabfhMYy9SYOhd8G3zq3Ag-tbeh_mhHRwFt-QV53pE749vCfk--XF7fl1dvPt6sv52U1mBVNTpqBhQpWFldw0TSOblgthKy4lygpkV6m2A7BKSWgA27LsLC9NKy3WopAMihPycZ87xvBjxjTpwSWLfW88LmU0K1klRSEK9n9UKC6ZYlwuKN-jNoaUInZ6jG4w8Ukz0KsivdWrIr0q0lDrRdGy9P6QPzcDtn9X_jhZgA8HwCRr-i4ab136x9WshLJai37ac7gcbucw6mQdeouti2gn3Qb3XI9fawuvyw</recordid><startdate>20140201</startdate><enddate>20140201</enddate><creator>Juránková, Jana</creator><creator>Hůrková-Hofmannová, Lada</creator><creator>Volf, Jiří</creator><creator>Baláž, Vojtech</creator><creator>Piálek, Jaroslav</creator><general>Elsevier GmbH</general><general>Elsevier</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>7TM</scope><scope>M7N</scope></search><sort><creationdate>20140201</creationdate><title>Efficacy of magnetic capture in comparison with conventional DNA isolation in a survey of Toxoplasma gondii in wild house mice</title><author>Juránková, Jana ; Hůrková-Hofmannová, Lada ; Volf, Jiří ; Baláž, Vojtech ; Piálek, Jaroslav</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c419t-90b14953c72abbb7bd244c6277e7607f69df00c9970b0ed55fc25ad7ce8437103</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Animal protozoal diseases</topic><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>DNA, Protozoan - analysis</topic><topic>DNA, Protozoan - isolation & purification</topic><topic>Female</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Genetic Techniques - standards</topic><topic>House mice</topic><topic>Infectious diseases</topic><topic>Life cycle. Host-agent relationship. Pathogenesis</topic><topic>Limit of Detection</topic><topic>Magnetic capture</topic><topic>Magnetic Phenomena</topic><topic>Medical sciences</topic><topic>Mice</topic><topic>Mus musculus</topic><topic>Parasitic diseases</topic><topic>Polymerase Chain Reaction - standards</topic><topic>Protozoa</topic><topic>Protozoal diseases</topic><topic>Quantitative PCR</topic><topic>Toxoplasma - genetics</topic><topic>Toxoplasma gondii</topic><topic>Toxoplasmosis, Animal - diagnosis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Juránková, Jana</creatorcontrib><creatorcontrib>Hůrková-Hofmannová, Lada</creatorcontrib><creatorcontrib>Volf, Jiří</creatorcontrib><creatorcontrib>Baláž, Vojtech</creatorcontrib><creatorcontrib>Piálek, Jaroslav</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>Nucleic Acids Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><jtitle>European journal of protistology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Juránková, Jana</au><au>Hůrková-Hofmannová, Lada</au><au>Volf, Jiří</au><au>Baláž, Vojtech</au><au>Piálek, Jaroslav</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Efficacy of magnetic capture in comparison with conventional DNA isolation in a survey of Toxoplasma gondii in wild house mice</atitle><jtitle>European journal of protistology</jtitle><addtitle>Eur J Protistol</addtitle><date>2014-02-01</date><risdate>2014</risdate><volume>50</volume><issue>1</issue><spage>11</spage><epage>15</epage><pages>11-15</pages><issn>0932-4739</issn><eissn>1618-0429</eissn><abstract>Toxoplasma gondii is a zoonotic parasite with a world-wide distribution. House mice (Mus musculus) play an important role as a reservoir host in the parasite life cycle. However, their detection in mouse brain is limited because the host potentially harbours only a few tissue cysts. In order to improve the diagnosis, we tested a novel protocol for T. gondii detection in mice and compared this technique to a standard PCR-based protocol using a commercial kit for DNA isolation. Efficacy of magnetic capture for isolation of T. gondii DNA from whole host brains was tested in brain samples of laboratory mice spiked with 1 up to 104 tachyzoites. Real-time PCR revealed that even 1–5 tachyzoites can be detected after magnetic capture. Also this method is suitable to quantify parasite numbers in mouse brains with more than 10 tachyzoite equivalents. To assess the two techniques in wild mice, we employed a dataset consisting of 243 individuals. 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subjects | Animal protozoal diseases Animals Biological and medical sciences DNA, Protozoan - analysis DNA, Protozoan - isolation & purification Female Fundamental and applied biological sciences. Psychology Genetic Techniques - standards House mice Infectious diseases Life cycle. Host-agent relationship. Pathogenesis Limit of Detection Magnetic capture Magnetic Phenomena Medical sciences Mice Mus musculus Parasitic diseases Polymerase Chain Reaction - standards Protozoa Protozoal diseases Quantitative PCR Toxoplasma - genetics Toxoplasma gondii Toxoplasmosis, Animal - diagnosis |
title | Efficacy of magnetic capture in comparison with conventional DNA isolation in a survey of Toxoplasma gondii in wild house mice |
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