Sensitivity of Polymerase Chain Reaction Assay for Rickettsia tsutsugamushi in Patients' Blood Samples
We developed a nested polymerase chain reaction (PCR) method to detect Rickettsia tsutsugamushi (R. tsutsugamushi) DNA and determined its sensitivity. Primers were selected from the DNA sequence of the 58-kDa group-specific antigen gene of the Karp strain. The target sequence of rickettsial DNA was...
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| Veröffentlicht in: | MICROBIOLOGY and IMMUNOLOGY 1992, Vol.36(11), pp.1145-1153 |
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| container_title | MICROBIOLOGY and IMMUNOLOGY |
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| creator | Murai, Koichi Tachibana, Nobuyoshi Okayama, Akihiko Shishime, Eiichi Tsuda, Kazunori Oshikawa, Tatsumi |
| description | We developed a nested polymerase chain reaction (PCR) method to detect Rickettsia tsutsugamushi (R. tsutsugamushi) DNA and determined its sensitivity. Primers were selected from the DNA sequence of the 58-kDa group-specific antigen gene of the Karp strain. The target sequence of rickettsial DNA was detectable as the band corresponding to 88bp in 1.0μg of the DNA extracted from BS-C-1 cells infected with R. tsutsugamushi. Rickettsia-specific bands were observed not only for the homologous Karp strain, but also for four heterologous strains: two other reference strains (Gilliam and Kato) and two prototype strains prevalent in Miyazaki district (Irie and Hirano). The minimum copy number detectable by this method was estimated to be five rickettsiae. All of nine peripheral blood mononuclear cell samples from patients with tsutsugamushi disease who were seen 2-11 days after disease onset tested positive for rickettsial DNA. The PCR assay method presented here could be a specific diagnostic tool for tsutsugamushi disease, especially in its early acute stage. |
| doi_str_mv | 10.1111/j.1348-0421.1992.tb02118.x |
| format | Article |
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Primers were selected from the DNA sequence of the 58-kDa group-specific antigen gene of the Karp strain. The target sequence of rickettsial DNA was detectable as the band corresponding to 88bp in 1.0μg of the DNA extracted from BS-C-1 cells infected with R. tsutsugamushi. Rickettsia-specific bands were observed not only for the homologous Karp strain, but also for four heterologous strains: two other reference strains (Gilliam and Kato) and two prototype strains prevalent in Miyazaki district (Irie and Hirano). The minimum copy number detectable by this method was estimated to be five rickettsiae. All of nine peripheral blood mononuclear cell samples from patients with tsutsugamushi disease who were seen 2-11 days after disease onset tested positive for rickettsial DNA. The PCR assay method presented here could be a specific diagnostic tool for tsutsugamushi disease, especially in its early acute stage.</description><identifier>ISSN: 0385-5600</identifier><identifier>EISSN: 1348-0421</identifier><identifier>DOI: 10.1111/j.1348-0421.1992.tb02118.x</identifier><identifier>PMID: 1491618</identifier><identifier>CODEN: MIIMDV</identifier><language>eng</language><publisher>Tokyo: Blackwell Publishing Ltd</publisher><subject>Adult ; Aged ; Aged, 80 and over ; Animals ; Antibodies, Bacterial - blood ; Bacteriological methods and techniques used in bacteriology ; Bacteriology ; Base Sequence ; Biological and medical sciences ; Cells, Cultured ; DNA, Bacterial - blood ; DNA, Bacterial - chemistry ; Female ; Fundamental and applied biological sciences. Psychology ; Humans ; Leukocytes, Mononuclear - microbiology ; Male ; Mice ; Mice, Nude ; Microbiology ; Middle Aged ; Molecular Sequence Data ; Orientia tsutsugamushi - genetics ; Orientia tsutsugamushi - immunology ; Orientia tsutsugamushi - isolation & purification ; Polymerase Chain Reaction ; Rickettsia ; Scrub Typhus - diagnosis ; Sensitivity and Specificity</subject><ispartof>MICROBIOLOGY and IMMUNOLOGY, 1992, Vol.36(11), pp.1145-1153</ispartof><rights>Center for Academic Publications Japan</rights><rights>owned by Center for Academic Publications Japan (Publisher)</rights><rights>1993 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c7068-8748f9d29e026ea335dc0310b60d6a3f6aba7bea5c745fd968356f4f09a969db3</citedby><cites>FETCH-LOGICAL-c7068-8748f9d29e026ea335dc0310b60d6a3f6aba7bea5c745fd968356f4f09a969db3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,1876,27903,27904</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=4555953$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/1491618$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Murai, Koichi</creatorcontrib><creatorcontrib>Tachibana, Nobuyoshi</creatorcontrib><creatorcontrib>Okayama, Akihiko</creatorcontrib><creatorcontrib>Shishime, Eiichi</creatorcontrib><creatorcontrib>Tsuda, Kazunori</creatorcontrib><creatorcontrib>Oshikawa, Tatsumi</creatorcontrib><title>Sensitivity of Polymerase Chain Reaction Assay for Rickettsia tsutsugamushi in Patients' Blood Samples</title><title>MICROBIOLOGY and IMMUNOLOGY</title><addtitle>Microbiology and Immunology</addtitle><description>We developed a nested polymerase chain reaction (PCR) method to detect Rickettsia tsutsugamushi (R. tsutsugamushi) DNA and determined its sensitivity. Primers were selected from the DNA sequence of the 58-kDa group-specific antigen gene of the Karp strain. The target sequence of rickettsial DNA was detectable as the band corresponding to 88bp in 1.0μg of the DNA extracted from BS-C-1 cells infected with R. tsutsugamushi. Rickettsia-specific bands were observed not only for the homologous Karp strain, but also for four heterologous strains: two other reference strains (Gilliam and Kato) and two prototype strains prevalent in Miyazaki district (Irie and Hirano). The minimum copy number detectable by this method was estimated to be five rickettsiae. All of nine peripheral blood mononuclear cell samples from patients with tsutsugamushi disease who were seen 2-11 days after disease onset tested positive for rickettsial DNA. The PCR assay method presented here could be a specific diagnostic tool for tsutsugamushi disease, especially in its early acute stage.</description><subject>Adult</subject><subject>Aged</subject><subject>Aged, 80 and over</subject><subject>Animals</subject><subject>Antibodies, Bacterial - blood</subject><subject>Bacteriological methods and techniques used in bacteriology</subject><subject>Bacteriology</subject><subject>Base Sequence</subject><subject>Biological and medical sciences</subject><subject>Cells, Cultured</subject><subject>DNA, Bacterial - blood</subject><subject>DNA, Bacterial - chemistry</subject><subject>Female</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Humans</subject><subject>Leukocytes, Mononuclear - microbiology</subject><subject>Male</subject><subject>Mice</subject><subject>Mice, Nude</subject><subject>Microbiology</subject><subject>Middle Aged</subject><subject>Molecular Sequence Data</subject><subject>Orientia tsutsugamushi - genetics</subject><subject>Orientia tsutsugamushi - immunology</subject><subject>Orientia tsutsugamushi - isolation & purification</subject><subject>Polymerase Chain Reaction</subject><subject>Rickettsia</subject><subject>Scrub Typhus - diagnosis</subject><subject>Sensitivity and Specificity</subject><issn>0385-5600</issn><issn>1348-0421</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1992</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqVkUtvEzEURkcIVErhJyBZCAGbCX57zIoS0QdqQ9UWsbTueDyN03mEsQPJv8dhosAKhGXZlu65x7a-LHtB8ISk8XYxIYwXOeaUTIjWdBJLTAkpJusH2eG-9DA7xKwQuZAYP86ehLDAmCpa8IPsgHBNJCkOs_rGdcFH_93HDeprdNU3m9YNEByazsF36NqBjb7v0HEIsEF1P6Brb-9djMEDimGV5h20qzD3KOFXEL3rYniNPjR9X6EbaJeNC0-zRzU0wT3b7UfZl5OPt9Oz_OLz6fn0-CK3CssiLxQval1R7TCVDhgTlcWM4FLiSgKrJZSgSgfCKi7qSsuCCVnzGmvQUlclO8pejd7l0H9buRBN64N1TQOd61fBKMYlZlr8EyRSckGpTOCbv4NcJ4pQyhP6bkTt0IcwuNosB9_CsDEEm21wZmG26ZhtOmYbnNkFZ9ap-fnunlXZuup365hUqr_c1SFYaOoBOuvDHuNCiPSthL0fsR--cZv_eIC5PL_8dUyKs1GxCBHu3N4BQ_S2caaFrvJEK2WYTNbdSggXe8TOYTCuS6p8VPkQ3foP072Riilhvs5OjcICz05mt-YT-wlGRN3P</recordid><startdate>19920101</startdate><enddate>19920101</enddate><creator>Murai, Koichi</creator><creator>Tachibana, Nobuyoshi</creator><creator>Okayama, Akihiko</creator><creator>Shishime, Eiichi</creator><creator>Tsuda, Kazunori</creator><creator>Oshikawa, Tatsumi</creator><general>Blackwell Publishing Ltd</general><general>Center For Academic Publications Japan</general><general>Center for Academic Publications Japan</general><scope>BSCLL</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>C1K</scope><scope>7X8</scope></search><sort><creationdate>19920101</creationdate><title>Sensitivity of Polymerase Chain Reaction Assay for Rickettsia tsutsugamushi in Patients' Blood Samples</title><author>Murai, Koichi ; Tachibana, Nobuyoshi ; Okayama, Akihiko ; Shishime, Eiichi ; Tsuda, Kazunori ; Oshikawa, Tatsumi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c7068-8748f9d29e026ea335dc0310b60d6a3f6aba7bea5c745fd968356f4f09a969db3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1992</creationdate><topic>Adult</topic><topic>Aged</topic><topic>Aged, 80 and over</topic><topic>Animals</topic><topic>Antibodies, Bacterial - blood</topic><topic>Bacteriological methods and techniques used in bacteriology</topic><topic>Bacteriology</topic><topic>Base Sequence</topic><topic>Biological and medical sciences</topic><topic>Cells, Cultured</topic><topic>DNA, Bacterial - blood</topic><topic>DNA, Bacterial - chemistry</topic><topic>Female</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Humans</topic><topic>Leukocytes, Mononuclear - microbiology</topic><topic>Male</topic><topic>Mice</topic><topic>Mice, Nude</topic><topic>Microbiology</topic><topic>Middle Aged</topic><topic>Molecular Sequence Data</topic><topic>Orientia tsutsugamushi - genetics</topic><topic>Orientia tsutsugamushi - immunology</topic><topic>Orientia tsutsugamushi - isolation & purification</topic><topic>Polymerase Chain Reaction</topic><topic>Rickettsia</topic><topic>Scrub Typhus - diagnosis</topic><topic>Sensitivity and Specificity</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Murai, Koichi</creatorcontrib><creatorcontrib>Tachibana, Nobuyoshi</creatorcontrib><creatorcontrib>Okayama, Akihiko</creatorcontrib><creatorcontrib>Shishime, Eiichi</creatorcontrib><creatorcontrib>Tsuda, Kazunori</creatorcontrib><creatorcontrib>Oshikawa, Tatsumi</creatorcontrib><collection>Istex</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>Environmental Sciences and Pollution Management</collection><collection>MEDLINE - Academic</collection><jtitle>MICROBIOLOGY and IMMUNOLOGY</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Murai, Koichi</au><au>Tachibana, Nobuyoshi</au><au>Okayama, Akihiko</au><au>Shishime, Eiichi</au><au>Tsuda, Kazunori</au><au>Oshikawa, Tatsumi</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Sensitivity of Polymerase Chain Reaction Assay for Rickettsia tsutsugamushi in Patients' Blood Samples</atitle><jtitle>MICROBIOLOGY and IMMUNOLOGY</jtitle><addtitle>Microbiology and Immunology</addtitle><date>1992-01-01</date><risdate>1992</risdate><volume>36</volume><issue>11</issue><spage>1145</spage><epage>1153</epage><pages>1145-1153</pages><issn>0385-5600</issn><eissn>1348-0421</eissn><coden>MIIMDV</coden><abstract>We developed a nested polymerase chain reaction (PCR) method to detect Rickettsia tsutsugamushi (R. tsutsugamushi) DNA and determined its sensitivity. Primers were selected from the DNA sequence of the 58-kDa group-specific antigen gene of the Karp strain. The target sequence of rickettsial DNA was detectable as the band corresponding to 88bp in 1.0μg of the DNA extracted from BS-C-1 cells infected with R. tsutsugamushi. Rickettsia-specific bands were observed not only for the homologous Karp strain, but also for four heterologous strains: two other reference strains (Gilliam and Kato) and two prototype strains prevalent in Miyazaki district (Irie and Hirano). The minimum copy number detectable by this method was estimated to be five rickettsiae. All of nine peripheral blood mononuclear cell samples from patients with tsutsugamushi disease who were seen 2-11 days after disease onset tested positive for rickettsial DNA. The PCR assay method presented here could be a specific diagnostic tool for tsutsugamushi disease, especially in its early acute stage.</abstract><cop>Tokyo</cop><pub>Blackwell Publishing Ltd</pub><pmid>1491618</pmid><doi>10.1111/j.1348-0421.1992.tb02118.x</doi><tpages>9</tpages><oa>free_for_read</oa></addata></record> |
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| identifier | ISSN: 0385-5600 |
| ispartof | MICROBIOLOGY and IMMUNOLOGY, 1992, Vol.36(11), pp.1145-1153 |
| issn | 0385-5600 1348-0421 |
| language | eng |
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| source | J-STAGE Free; MEDLINE; Freely Accessible Japanese Titles; Alma/SFX Local Collection |
| subjects | Adult Aged Aged, 80 and over Animals Antibodies, Bacterial - blood Bacteriological methods and techniques used in bacteriology Bacteriology Base Sequence Biological and medical sciences Cells, Cultured DNA, Bacterial - blood DNA, Bacterial - chemistry Female Fundamental and applied biological sciences. Psychology Humans Leukocytes, Mononuclear - microbiology Male Mice Mice, Nude Microbiology Middle Aged Molecular Sequence Data Orientia tsutsugamushi - genetics Orientia tsutsugamushi - immunology Orientia tsutsugamushi - isolation & purification Polymerase Chain Reaction Rickettsia Scrub Typhus - diagnosis Sensitivity and Specificity |
| title | Sensitivity of Polymerase Chain Reaction Assay for Rickettsia tsutsugamushi in Patients' Blood Samples |
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