{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,3]],"date-time":"2026-10-03T20:09:17Z","timestamp":1791058157723,"version":"4.1.0"},"reference-count":30,"publisher":"American Society for Microbiology","issue":"2","license":[{"start":{"date-parts":[[2003,2,1]],"date-time":"2003-02-01T00:00:00Z","timestamp":1044057600000},"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":[[2003,2]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            Human granulocytic ehrlichiosis is an emerging infectious disease in the United States and Europe, and PCR methods have been shown to be effective for the diagnosis of acute infections. Numerous PCR assays and primer sets have been reported in the literature. The analytical sensitivities (limits of detection) of 13 published PCR primer sets were compared using DNA extracted from serial dilutions of\n            <jats:italic>Anaplasma phagocytophilum<\/jats:italic>\n            -infected HL-60 cells. The specificity of the assays that were able to detect \u22642.5 infected cells was tested by the use of template DNA extracted from\n            <jats:italic>Ehrlichia chaffeensis<\/jats:italic>\n            ,\n            <jats:italic>Rickettsia rickettsii<\/jats:italic>\n            , and\n            <jats:italic>Bartonella henselae<\/jats:italic>\n            . The assays with the lowest limits of detection were shown to be a nested assay that amplifies the 16S rRNA gene (primer pairs ge3a-ge10 [primary] and ge9-ge3 [nested]; detects 0.25 infected cell), a direct assay that amplifies the major surface protein gene\n            <jats:italic>msp2<\/jats:italic>\n            (primer pair msp2-3f-msp2-3r; detects 0.25 infected cell), and a direct assay that amplifies the 16S rRNA gene (primer pair ehr521-ehr790; detects 0.25 infected cell). The specificity and limit of detection of the MSP2 and 16S rRNA direct assays were further tested by use of\n            <jats:italic>A. phagocytophilum<\/jats:italic>\n            template DNA from both North America and Europe and from human, tick, white-footed mouse, equine, deer, bovine, and wood rat samples and of template DNA from closely related species (\n            <jats:italic>Anaplasma marginale<\/jats:italic>\n            , the white-tailed deer agent, and additional\n            <jats:italic>E. chaffeensis<\/jats:italic>\n            -positive samples). Three manufacturers' PCR kits were tested and showed distinct variations in the limit of detection, specificity, and nonspecific background amplification. The importance of these results for the molecular diagnosis of human granulocytic ehrlichiosis is discussed.\n          <\/jats:p>","DOI":"10.1128\/jcm.41.2.717-722.2003","type":"journal-article","created":{"date-parts":[[2003,2,6]],"date-time":"2003-02-06T20:03:31Z","timestamp":1044561811000},"page":"717-722","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":107,"title":["Comparison of PCR Assays for Detection of the Agent of Human Granulocytic Ehrlichiosis,\n            <i>Anaplasma phagocytophilum<\/i>"],"prefix":"10.1128","volume":"41","author":[{"given":"Robert F.","family":"Massung","sequence":"first","affiliation":[{"name":"Viral and Rickettsial Zoonoses Branch, Division of Viral and Rickettsial Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kimetha G.","family":"Slater","sequence":"additional","affiliation":[{"name":"Viral and Rickettsial Zoonoses Branch, Division of Viral and Rickettsial Diseases, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.38.2.635-638.2000"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.29.12.2838-2842.1991"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1001\/jama.1994.03520030054028"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1080\/00034983.1994.11812872"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.36.12.2799"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM199907153410303"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.32.3.589-595.1994"},{"key":"e_1_3_2_9_2","unstructured":"Childs J. E. and C. D. Paddock. 2002. Ehrlichia species (the ehrlichioses) p. 913-916. In S. S. Long L. K. Pickering and C. G. Prober (ed.) Principles and practice of pediatric infectious diseases 2nd ed. Churchill Livingstone Philadelphia Pa."},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.29.12.2741-2745.1991"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.2307\/3284115"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1093\/clinids\/20.5.1102"},{"key":"e_1_3_2_13_2","first-page":"2145","year":"2001","unstructured":"Dumler, J. S., A. F. Barbet, C. P. J. Bekker, G. A. Dasch, G. H. Palmer, S. C. Ray, Y. Rikihisa, and F. R. Rurangirwa. 2001. Reorganization of genera in the families Rickettsiaceae and Anaplasmataceae in the order Rickettsiales: unification of some species of Ehrlichia with Anaplasma, Cowdria with Ehrlichia and Ehrlichia with Neorickettsia, descriptions of six new species combinations and designation of Ehrlichia equi and \u2032HGE agent' as subjective synonyms of Ehrlichia phagocytophila. Int. J. Syst. Evol. Microbiol. 51:2145-2165.","journal-title":"Int. J. Syst. E"},{"key":"e_1_3_2_14_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM199601253340401"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.1128\/IAI.66.7.3264-3269.1998"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.41.1.76"},{"key":"e_1_3_2_17_2","unstructured":"Kolbert C. 1996. Detection of the agent of human granulocytic ehrlichiosis by PCR p.106-111. In D. Persing (ed.) 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