{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,11,5]],"date-time":"2025-11-05T06:36:57Z","timestamp":1762324617058},"reference-count":45,"publisher":"American Society for Microbiology","issue":"4","license":[{"start":{"date-parts":[[1998,7,1]],"date-time":"1998-07-01T00:00:00Z","timestamp":899251200000},"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":["Clin Diagn Lab Immunol"],"published-print":{"date-parts":[[1998,7]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            The variable 56-kDa major outer membrane protein of\n            <jats:italic>Orientia tsutsugamushi<\/jats:italic>\n            is the immunodominant antigen in human scrub typhus infections. The gene encoding this protein from Karp strain was cloned into the expression vector pET11a. The recombinant protein (r56) was expressed as a truncated nonfusion protein (amino acids 80 to 456 of the open reading frame) which formed an inclusion body when expressed in\n            <jats:italic>Escherichia coli<\/jats:italic>\n            BL21. Refolded r56 was purified and compared to purified whole-cell lysate of the Karp strain of\n            <jats:italic>O. tsutsugamushi<\/jats:italic>\n            by immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA) for reactivity with rabbit sera prepared against eight antigenic prototypes of\n            <jats:italic>O. tsutsugamushi<\/jats:italic>\n            as well as several other species of\n            <jats:italic>Rickettsiales<\/jats:italic>\n            and nonrickettsial antigens. Refolded r56 exhibited broad reactivity with the rabbit antisera against the\n            <jats:italic>Orientia<\/jats:italic>\n            prototypes, and the ELISA reactions with the r56 and Karp whole-cell lysate antigens correlated well (\n            <jats:italic>r<\/jats:italic>\n            = 0.81,\n            <jats:italic>n<\/jats:italic>\n            = 22, sensitivity compared to that of standard ELISA of 91%). Refolded r56 did not react with most antisera against other rickettsial species or control antigens (specificity = 92%,\n            <jats:italic>n<\/jats:italic>\n            = 13) using a positive cutoff value determined with eight uninfected rabbit sera. Refolded r56 was evaluated further by ELISA, using 128 sera obtained from patients with suspected scrub typhus from Korat, Thailand, and 74 serum specimens from healthy Thai soldiers. By using the indirect immunoperoxidase assay as the reference assay, the recombinant antigen exhibited a sensitivity and specificity of 93% or greater for detection of both IgG and IgM in the ELISA at 1:400 serum dilution. These results strongly suggest that purified r56 is a suitable candidate for replacing the density gradient-purified, rickettsia-derived, whole-cell antigen currently used in the commercial dipstick assay available in the United States.\n          <\/jats:p>","DOI":"10.1128\/cdli.5.4.519-526.1998","type":"journal-article","created":{"date-parts":[[2019,12,31]],"date-time":"2019-12-31T19:21:07Z","timestamp":1577820067000},"page":"519-526","update-policy":"http:\/\/dx.doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":40,"title":["Expression and Refolding of Truncated Recombinant Major Outer Membrane Protein Antigen (r56) of\n            <i>Orientia tsutsugamushi<\/i>\n            and Its Use in Enzyme-Linked Immunosorbent Assays"],"prefix":"10.1128","volume":"5","author":[{"given":"W.-M.","family":"Ching","sequence":"first","affiliation":[{"name":"<!--label omitted: 1-->Viral and Rickettsial Diseases Program, Infectious Diseases Department, Naval Medical Research Institute, Bethesda, Maryland1;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"H.","family":"Wang","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->Viral and Rickettsial Diseases Program, Infectious Diseases Department, Naval Medical Research Institute, Bethesda, Maryland1;"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"C.","family":"Eamsila","sequence":"additional","affiliation":[{"name":"<!--label omitted: 2-->Thai Component, Armed Forces Research Institute of Medical Science, Bangkok, Thailand2; and"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"D. J.","family":"Kelly","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->Viral and Rickettsial Diseases Program, Infectious Diseases Department, Naval Medical Research Institute, Bethesda, Maryland1;"},{"name":"<!--label omitted: 3-->Division of Communicable Diseases and Immunology, Walter Reed Army Institute of Research, Washington, D.C.3"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"G. A.","family":"Dasch","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->Viral and Rickettsial Diseases Program, Infectious Diseases Department, Naval Medical Research Institute, Bethesda, Maryland1;"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1128\/iai.55.9.2290-2292.1987"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1101\/SQB.1984.049.01.057"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.4269\/ajtmh.1982.31.532"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.3181\/00379727-112-28107"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/0035-9203(76)90127-9"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/0035-9203(78)90138-4"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.3346\/jkms.1995.10.4.227"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/0161-5890(92)90161-P"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1021\/bi00290a005"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.11.6.584-588.1980"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.9.1.38-48.1979"},{"key":"e_1_3_2_13_2","unstructured":"Dasch\nG. 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