Genotyping of single Cryptosporidium oocysts in sewage by semi-nested PCR and direct sequencing
This study describes an approach for genotyping individual Cryptosporidium oocysts obtained from sewage. We isolated single immunofluorescent assay (IFA)-stained Cryptosporidium oocysts from sewage concentrate using glass capillary pipettes and inverted epifluorescence microscopy. Each isolated Cryp...
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| Veröffentlicht in: | Water research (Oxford) 2006-07, Vol.40 (13), p.2527-2532 |
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| creator | Hashimoto, Atsushi Sugimoto, Hitomi Morita, Shigemitsu Hirata, Tsuyoshi |
| description | This study describes an approach for genotyping individual
Cryptosporidium oocysts obtained from sewage. We isolated single immunofluorescent assay (IFA)-stained
Cryptosporidium oocysts from sewage concentrate using glass capillary pipettes and inverted epifluorescence microscopy. Each isolated
Cryptosporidium oocyst was analyzed by semi-nested PCR for the 18S rRNA gene and direct sequencing of the PCR products. A total of 74 of 107 oocysts isolated from sewage were genotyped successfully. Of the 74 genotyped isolates, 51% (38 oocysts) were identified as
C. parvum genotype 1, 4% (3 oocysts) of
C. parvum VF383 human isolates, 20% (15 oocysts) of
C. parvum genotype 2, 14% (10 oocysts) of
C. meleagridis, 7% (5 oocysts) of
C. sp. Pig 1, 3% (2 oocysts) of
C. sp PG1-26 pig isolates and 1% (1 oocyst) of
C. parvum CPM1 isolated from mouse. The results of this study demonstrate that 18S rRNA-based semi-nested PCR and direct sequencing can be used to characterize individual
Cryptosporidium oocysts and also to reveal the distribution of
Cryptosporidium genotypes in environmental waters. |
| doi_str_mv | 10.1016/j.watres.2006.04.038 |
| format | Article |
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Cryptosporidium oocysts obtained from sewage. We isolated single immunofluorescent assay (IFA)-stained
Cryptosporidium oocysts from sewage concentrate using glass capillary pipettes and inverted epifluorescence microscopy. Each isolated
Cryptosporidium oocyst was analyzed by semi-nested PCR for the 18S rRNA gene and direct sequencing of the PCR products. A total of 74 of 107 oocysts isolated from sewage were genotyped successfully. Of the 74 genotyped isolates, 51% (38 oocysts) were identified as
C. parvum genotype 1, 4% (3 oocysts) of
C. parvum VF383 human isolates, 20% (15 oocysts) of
C. parvum genotype 2, 14% (10 oocysts) of
C. meleagridis, 7% (5 oocysts) of
C. sp. Pig 1, 3% (2 oocysts) of
C. sp PG1-26 pig isolates and 1% (1 oocyst) of
C. parvum CPM1 isolated from mouse. The results of this study demonstrate that 18S rRNA-based semi-nested PCR and direct sequencing can be used to characterize individual
Cryptosporidium oocysts and also to reveal the distribution of
Cryptosporidium genotypes in environmental waters.</description><identifier>ISSN: 0043-1354</identifier><identifier>EISSN: 1879-2448</identifier><identifier>DOI: 10.1016/j.watres.2006.04.038</identifier><identifier>PMID: 16790257</identifier><language>eng</language><publisher>England: Elsevier Ltd</publisher><subject>18S rRNA ; animal parasites and pests ; Animals ; Cryptosporidium ; Cryptosporidium meleagridis ; Cryptosporidium parvum ; Cryptosporidium parvum - genetics ; fluorescent antibody technique ; genes ; Genotype ; microscopy ; molecular epidemiology ; Oocysts ; parasites ; pathogen identification ; PCR ; polymerase chain reaction ; Polymerase Chain Reaction - methods ; ribosomal RNA ; RNA, Ribosomal, 18S - genetics ; Sensitivity and Specificity ; sequence analysis ; Sequence Analysis, DNA - methods ; Sewage ; Sewage - parasitology ; water pollution</subject><ispartof>Water research (Oxford), 2006-07, Vol.40 (13), p.2527-2532</ispartof><rights>2006 Elsevier Ltd</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c481t-db446973317dd2ef03876426fc34df322991e8530e16954f152a4c963756402a3</citedby><cites>FETCH-LOGICAL-c481t-db446973317dd2ef03876426fc34df322991e8530e16954f152a4c963756402a3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.watres.2006.04.038$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/16790257$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Hashimoto, Atsushi</creatorcontrib><creatorcontrib>Sugimoto, Hitomi</creatorcontrib><creatorcontrib>Morita, Shigemitsu</creatorcontrib><creatorcontrib>Hirata, Tsuyoshi</creatorcontrib><title>Genotyping of single Cryptosporidium oocysts in sewage by semi-nested PCR and direct sequencing</title><title>Water research (Oxford)</title><addtitle>Water Res</addtitle><description>This study describes an approach for genotyping individual
Cryptosporidium oocysts obtained from sewage. We isolated single immunofluorescent assay (IFA)-stained
Cryptosporidium oocysts from sewage concentrate using glass capillary pipettes and inverted epifluorescence microscopy. Each isolated
Cryptosporidium oocyst was analyzed by semi-nested PCR for the 18S rRNA gene and direct sequencing of the PCR products. A total of 74 of 107 oocysts isolated from sewage were genotyped successfully. Of the 74 genotyped isolates, 51% (38 oocysts) were identified as
C. parvum genotype 1, 4% (3 oocysts) of
C. parvum VF383 human isolates, 20% (15 oocysts) of
C. parvum genotype 2, 14% (10 oocysts) of
C. meleagridis, 7% (5 oocysts) of
C. sp. Pig 1, 3% (2 oocysts) of
C. sp PG1-26 pig isolates and 1% (1 oocyst) of
C. parvum CPM1 isolated from mouse. The results of this study demonstrate that 18S rRNA-based semi-nested PCR and direct sequencing can be used to characterize individual
Cryptosporidium oocysts and also to reveal the distribution of
Cryptosporidium genotypes in environmental waters.</description><subject>18S rRNA</subject><subject>animal parasites and pests</subject><subject>Animals</subject><subject>Cryptosporidium</subject><subject>Cryptosporidium meleagridis</subject><subject>Cryptosporidium parvum</subject><subject>Cryptosporidium parvum - genetics</subject><subject>fluorescent antibody technique</subject><subject>genes</subject><subject>Genotype</subject><subject>microscopy</subject><subject>molecular epidemiology</subject><subject>Oocysts</subject><subject>parasites</subject><subject>pathogen identification</subject><subject>PCR</subject><subject>polymerase chain reaction</subject><subject>Polymerase Chain Reaction - methods</subject><subject>ribosomal RNA</subject><subject>RNA, Ribosomal, 18S - genetics</subject><subject>Sensitivity and Specificity</subject><subject>sequence analysis</subject><subject>Sequence Analysis, DNA - methods</subject><subject>Sewage</subject><subject>Sewage - parasitology</subject><subject>water pollution</subject><issn>0043-1354</issn><issn>1879-2448</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2006</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kE1r3DAQhkVpabZJ_kFpdcrN7ujDkn0plKVNCoGWtjkLrTRetKwtV_I2-N9HwQu95TSCeead0UPIewY1A6Y-HepHOyfMNQdQNcgaRPuKbFiru4pL2b4mGwApKiYaeUHe5XwAAM5F95ZcMKU74I3eEHOLY5yXKYx7GnuaSz0i3aZlmmOeYgo-nAYao1vynGkYacZHu0e6W8prCNWIeUZPf25_UTt66kNCN5fW3xOOroRdkTe9PWa8PtdL8vDt65_tXXX_4_b79st95WTL5srvpFSdFoJp7zn25S9aSa56J6TvBeddx7BtBCBTXSN71nArXaeEbpQEbsUluVlzpxTL7jybIWSHx6MdMZ6yYZoLLbUqoFxBl2LOCXszpTDYtBgG5lmsOZhVrHkWa0CackwZ-3DOP-0G9P-HziYL8HEFehuN3aeQzcNvDkwAg7aTghXi80pg8fAvYDLZhWIJV2nGx_DyDU-u0ZSF</recordid><startdate>20060701</startdate><enddate>20060701</enddate><creator>Hashimoto, Atsushi</creator><creator>Sugimoto, Hitomi</creator><creator>Morita, Shigemitsu</creator><creator>Hirata, Tsuyoshi</creator><general>Elsevier Ltd</general><scope>FBQ</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>7QH</scope><scope>7UA</scope><scope>8FD</scope><scope>C1K</scope><scope>F1W</scope><scope>FR3</scope><scope>H96</scope><scope>L.G</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope></search><sort><creationdate>20060701</creationdate><title>Genotyping of single Cryptosporidium oocysts in sewage by semi-nested PCR and direct sequencing</title><author>Hashimoto, Atsushi ; Sugimoto, Hitomi ; Morita, Shigemitsu ; Hirata, Tsuyoshi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c481t-db446973317dd2ef03876426fc34df322991e8530e16954f152a4c963756402a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2006</creationdate><topic>18S rRNA</topic><topic>animal parasites and pests</topic><topic>Animals</topic><topic>Cryptosporidium</topic><topic>Cryptosporidium meleagridis</topic><topic>Cryptosporidium parvum</topic><topic>Cryptosporidium parvum - genetics</topic><topic>fluorescent antibody technique</topic><topic>genes</topic><topic>Genotype</topic><topic>microscopy</topic><topic>molecular epidemiology</topic><topic>Oocysts</topic><topic>parasites</topic><topic>pathogen identification</topic><topic>PCR</topic><topic>polymerase chain reaction</topic><topic>Polymerase Chain Reaction - methods</topic><topic>ribosomal RNA</topic><topic>RNA, Ribosomal, 18S - genetics</topic><topic>Sensitivity and Specificity</topic><topic>sequence analysis</topic><topic>Sequence Analysis, DNA - methods</topic><topic>Sewage</topic><topic>Sewage - parasitology</topic><topic>water pollution</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Hashimoto, Atsushi</creatorcontrib><creatorcontrib>Sugimoto, Hitomi</creatorcontrib><creatorcontrib>Morita, Shigemitsu</creatorcontrib><creatorcontrib>Hirata, Tsuyoshi</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Aqualine</collection><collection>Water Resources 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>Aquatic Science & Fisheries Abstracts (ASFA) 2: Ocean Technology, Policy & Non-Living Resources</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>Genetics Abstracts</collection><jtitle>Water research (Oxford)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Hashimoto, Atsushi</au><au>Sugimoto, Hitomi</au><au>Morita, Shigemitsu</au><au>Hirata, Tsuyoshi</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Genotyping of single Cryptosporidium oocysts in sewage by semi-nested PCR and direct sequencing</atitle><jtitle>Water research (Oxford)</jtitle><addtitle>Water Res</addtitle><date>2006-07-01</date><risdate>2006</risdate><volume>40</volume><issue>13</issue><spage>2527</spage><epage>2532</epage><pages>2527-2532</pages><issn>0043-1354</issn><eissn>1879-2448</eissn><abstract>This study describes an approach for genotyping individual
Cryptosporidium oocysts obtained from sewage. We isolated single immunofluorescent assay (IFA)-stained
Cryptosporidium oocysts from sewage concentrate using glass capillary pipettes and inverted epifluorescence microscopy. Each isolated
Cryptosporidium oocyst was analyzed by semi-nested PCR for the 18S rRNA gene and direct sequencing of the PCR products. A total of 74 of 107 oocysts isolated from sewage were genotyped successfully. Of the 74 genotyped isolates, 51% (38 oocysts) were identified as
C. parvum genotype 1, 4% (3 oocysts) of
C. parvum VF383 human isolates, 20% (15 oocysts) of
C. parvum genotype 2, 14% (10 oocysts) of
C. meleagridis, 7% (5 oocysts) of
C. sp. Pig 1, 3% (2 oocysts) of
C. sp PG1-26 pig isolates and 1% (1 oocyst) of
C. parvum CPM1 isolated from mouse. The results of this study demonstrate that 18S rRNA-based semi-nested PCR and direct sequencing can be used to characterize individual
Cryptosporidium oocysts and also to reveal the distribution of
Cryptosporidium genotypes in environmental waters.</abstract><cop>England</cop><pub>Elsevier Ltd</pub><pmid>16790257</pmid><doi>10.1016/j.watres.2006.04.038</doi><tpages>6</tpages></addata></record> |
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| subjects | 18S rRNA animal parasites and pests Animals Cryptosporidium Cryptosporidium meleagridis Cryptosporidium parvum Cryptosporidium parvum - genetics fluorescent antibody technique genes Genotype microscopy molecular epidemiology Oocysts parasites pathogen identification PCR polymerase chain reaction Polymerase Chain Reaction - methods ribosomal RNA RNA, Ribosomal, 18S - genetics Sensitivity and Specificity sequence analysis Sequence Analysis, DNA - methods Sewage Sewage - parasitology water pollution |
| title | Genotyping of single Cryptosporidium oocysts in sewage by semi-nested PCR and direct sequencing |
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