{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,13]],"date-time":"2026-10-13T12:02:01Z","timestamp":1791892921065,"version":"4.3.4"},"reference-count":25,"publisher":"American Society for Microbiology","issue":"2","license":[{"start":{"date-parts":[[2011,2,1]],"date-time":"2011-02-01T00:00:00Z","timestamp":1296518400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.asm.org\/non-commercial-tdm-license"}],"content-domain":{"domain":["journals.asm.org"],"crossmark-restriction":true},"short-container-title":["J Clin Microbiol"],"published-print":{"date-parts":[[2011,2]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    <jats:named-content content-type=\"genus-species\">Orientia tsutsugamushi<\/jats:named-content>\n                    is the causative agent of scrub typhus. For the diagnosis of scrub typhus, we investigated the performances of conventional PCR (C-PCR), nested PCR (N-PCR), and real-time quantitative PCR (Q-PCR) targeting the\n                    <jats:named-content content-type=\"genus-species\">O. tsutsugamushi<\/jats:named-content>\n                    -specific 47-kDa gene. To compare the detection sensitivities of the three techniques, we used two template systems that used plasmid DNA (plasmid detection sensitivity), including a partial region of the 47-kDa gene, and genomic DNA (genomic detection sensitivity) from a buffy coat sample of a single patient. The plasmid detection sensitivities of C-PCR, N-PCR, and Q-PCR were 5 \u00d7 10\n                    <jats:sup>4<\/jats:sup>\n                    copies\/\u03bcl, 5 copies\/\u03bcl, and 50 copies\/\u03bcl, respectively. The results of C-PCR, N-PCR, and Q-PCR performed with undiluted genomic DNA were negative, positive, and positive, respectively. The genomic detection sensitivities of N-PCR and Q-PCR were 64-fold and 16-fold (crossing point [Cp], 37.7; 426 copies\/\u03bcl), respectively. For relative quantification of\n                    <jats:named-content content-type=\"genus-species\">O. tsutsugamushi<\/jats:named-content>\n                    bacteria per volume of whole blood, we performed real-time DNA PCR analysis of the human\n                    <jats:italic>GAPDH<\/jats:italic>\n                    gene, along with the\n                    <jats:named-content content-type=\"genus-species\">O. tsutsugamushi<\/jats:named-content>\n                    47-kDa gene. At a 16-fold dilution, the copy number and genomic equivalent (GE) of\n                    <jats:italic>GAPDH<\/jats:italic>\n                    were 1.1 \u00d7 10\n                    <jats:sup>5<\/jats:sup>\n                    copies\/\u03bcl (Cp, 22.64) and 5.5 \u00d7 10\n                    <jats:sup>4<\/jats:sup>\n                    GEs\/\u03bcl, respectively. Therefore, the relative concentration of\n                    <jats:named-content content-type=\"genus-species\">O. tsutsugamushi<\/jats:named-content>\n                    at a 16-fold dilution was 0.0078 organism\/one white blood cell (WBC) and 117 organisms\/\u03bcl of whole blood, because the WBC count of the patient was 1.5 \u00d7 10\n                    <jats:sup>4<\/jats:sup>\n                    cells\/\u03bcl of whole blood. The sensitivities of C-PCR, N-PCR, and Q-PCR performed with blood samples taken from patients within 4 weeks of onset of fever were 7.3% (95% confidence interval [CI], 1.6 to 19.9), 85.4% (95% CI, 70.8 to 94.4), and 82.9% (95% CI, 67.9 to 92.8), respectively. All evaluated assays were 100% specific for\n                    <jats:named-content content-type=\"genus-species\">O. tsutsugamushi<\/jats:named-content>\n                    . In conclusion, given its combined sensitivity, specificity, and speed, Q-PCR is the preferred assay for the diagnosis of scrub typhus.\n                  <\/jats:p>","DOI":"10.1128\/jcm.01216-09","type":"journal-article","created":{"date-parts":[[2010,11,10]],"date-time":"2010-11-10T22:52:43Z","timestamp":1289429563000},"page":"607-612","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":92,"title":["Comparison of Conventional, Nested, and Real-Time Quantitative PCR for Diagnosis of Scrub Typhus"],"prefix":"10.1128","volume":"49","author":[{"given":"Dong-Min","family":"Kim","sequence":"first","affiliation":[{"name":"Division of Infectious Disease, Department of Internal Medicine, Chosun University School of Medicine, Gwang-ju, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Geon","family":"Park","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine, Chosun University School of Medicine, Gwang-ju, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hyong Sun","family":"Kim","sequence":"additional","affiliation":[{"name":"Division of Infectious Disease, Department of Internal Medicine, Chosun University School of Medicine, Gwang-ju, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joo Young","family":"Lee","sequence":"additional","affiliation":[{"name":"Division of Infectious Disease, Department of Internal Medicine, Chosun University School of Medicine, Gwang-ju, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ganesh Prasad","family":"Neupane","sequence":"additional","affiliation":[{"name":"Division of Infectious Disease, Department of Internal Medicine, Chosun University School of Medicine, Gwang-ju, Republic of Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephen","family":"Graves","sequence":"additional","affiliation":[{"name":"Australian Rickettsial Reference Laboratory, Geelong Hospital, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"John","family":"Stenos","sequence":"additional","affiliation":[{"name":"Australian Rickettsial Reference Laboratory, Geelong Hospital, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1086\/510585"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.3181\/00379727-112-28107"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.29.11.2628-2630.1991"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.4269\/ajtmh.1996.54.647"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.4269\/ajtmh.2004.70.351"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.00634-08"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.01460-06"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.00027-08"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1348-0421.2000.tb02495.x"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1055\/s-2000-9621"},{"key":"e_1_3_2_12_2","unstructured":"Reference deleted."},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.36.10.2887-2892.1998"},{"key":"e_1_3_2_14_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1348-0421.1992.tb02118.x"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)42337-X"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.mcp.2004.04.005"},{"key":"e_1_3_2_17_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.43.7.3390-3397.2005"},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0167-7012(03)00071-X"},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jaad.2004.03.036"},{"key":"e_1_3_2_20_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.42.10.4512-4518.2004"},{"key":"e_1_3_2_21_2","doi-asserted-by":"publisher","DOI":"10.1002\/1097-0339(200008)23:2<134::AID-DC14>3.0.CO;2-N"},{"key":"e_1_3_2_22_2","doi-asserted-by":"publisher","DOI":"10.1097\/00006454-199103000-00006"},{"key":"e_1_3_2_23_2","first-page":"780","article-title":"Septic shock secondary to scrub typhus: characteristics and complications","volume":"33","author":"Thap L. 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