{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,11]],"date-time":"2026-02-11T15:29:47Z","timestamp":1770823787652,"version":"3.50.1"},"reference-count":31,"publisher":"American Society for Microbiology","issue":"6","license":[{"start":{"date-parts":[[2002,6,1]],"date-time":"2002-06-01T00:00:00Z","timestamp":1022889600000},"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":[[2002,6]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            A 300-bp plasmid fragment of the hemagglutinin gene was used as target DNA to develop a rapid real-time LightCycler (Roche Applied Science, Indianapolis, Ind.) PCR assay for laboratory detection of smallpox virus. PCR primers and probes were designed specifically for detection of smallpox virus DNA, but all viruses of the genus\n            <jats:italic>Orthopoxvirus<\/jats:italic>\n            tested could be detected by use of the hemagglutinin gene target sequence. Base pair mismatches in the 204-bp amplicon allowed discrimination of cowpox virus (melting temperature [\n            <jats:italic>\n              T\n              <jats:sub>m<\/jats:sub>\n            <\/jats:italic>\n            ], 56.40\u00b0C), monkeypox virus (\n            <jats:italic>\n              T\n              <jats:sub>m<\/jats:sub>\n            <\/jats:italic>\n            , 56.24\u00b0C), and vaccinia virus (\n            <jats:italic>\n              T\n              <jats:sub>m<\/jats:sub>\n            <\/jats:italic>\n            , 56.72\u00b0C), including the Dryvax vaccine strain, from smallpox virus (\n            <jats:italic>\n              T\n              <jats:sub>m<\/jats:sub>\n            <\/jats:italic>\n            , 62.45\u00b0C) by melting curve analysis. The analytical sensitivity was 5 to 10 copies of target DNA per sample. The assay was specific for members of the genus\n            <jats:italic>Orthopoxvirus<\/jats:italic>\n            ; the DNAs of herpes simplex virus and varicella-zoster virus were not detected by the smallpox virus LightCycler PCR.\n          <\/jats:p>","DOI":"10.1128\/jcm.40.6.1985-1988.2002","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T10:09:29Z","timestamp":1027764569000},"page":"1985-1988","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":81,"title":["Detection of Smallpox Virus DNA by LightCycler PCR"],"prefix":"10.1128","volume":"40","author":[{"given":"Mark J.","family":"Espy","sequence":"first","affiliation":[{"name":"Division of Clinical Microbiology"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"suffix":"III","given":"Franklin R.","family":"Cockerill","sequence":"additional","affiliation":[{"name":"Division of Clinical Microbiology"},{"name":"Division of Infectious Diseases"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Richard F.","family":"Meyer","sequence":"additional","affiliation":[{"name":"Centers for Disease Control and Prevention, Atlanta, Georgia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael D.","family":"Bowen","sequence":"additional","affiliation":[{"name":"Centers for Disease Control and Prevention, Atlanta, Georgia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gregory A.","family":"Poland","sequence":"additional","affiliation":[{"name":"Division of General Internal Medicine, Mayo ClinicFoundation; Rochester, Minnesota"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ted L.","family":"Hadfield","sequence":"additional","affiliation":[{"name":"Armed Forces Institute of Pathology, Walter Reed Army Medical Center, Washington, D.C."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thomas F.","family":"Smith","sequence":"additional","affiliation":[{"name":"Division of Clinical Microbiology"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_1_2_2","first-page":"269.","volume":"409","year":"2001","unstructured":"Anonymous. 2001. Don't underestimate the enemy. Nature409:269.","journal-title":"Nature"},{"key":"e_1_3_1_3_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1369-5274(98)80121-5"},{"key":"e_1_3_1_4_2","doi-asserted-by":"publisher","DOI":"10.1017\/S0022172400023457"},{"key":"e_1_3_1_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0966-842X(00)01855-2"},{"key":"e_1_3_1_6_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM199808203390811"},{"key":"e_1_3_1_7_2","first-page":"1","volume":"50","year":"2001","unstructured":"Centers for Disease Control and Prevention. 2001. Vaccinia (smallpox) vaccine. Recommendations of the Advisory Committee on Immunization Practices (ACIP). Morb. Mortal. Wkly. Rep.50:1-25.","journal-title":"Morb. Mortal. Wkly. Rep."},{"key":"e_1_3_1_8_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.38.8.3116-3118.2000"},{"key":"e_1_3_1_9_2","first-page":"2233","volume":"36","year":"2001","unstructured":"Espy, M. J., P. N. Rys, A. D. Wold, J. R. Uhl, L. M. Sloan, G. D. Jenkins, D. M. Ilstrup, F. R. Cockerill, I. I. I., R. Patel, J. E. Rosenblatt, and T. F. Smith. 2001. Detection of herpes simplex virus DNA in genital and dermal specimens by LightCycler PCR after extraction using the IsoQuick, MagNA Pure, and BioRobot 9604 methods. J. Clin. Microbiol.36:2233-2236.","journal-title":"J. Clin. Microbiol."},{"key":"e_1_3_1_10_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.38.9.3187-3189.2000"},{"key":"e_1_3_1_11_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.38.2.795-799.2000"},{"key":"e_1_3_1_12_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.283.5406.1279"},{"key":"e_1_3_1_13_2","doi-asserted-by":"publisher","DOI":"10.1001\/jama.281.22.2127"},{"key":"e_1_3_1_14_2","first-page":"331","volume":"15","year":"1973","unstructured":"Higashi, N. 1973. Electron microscopy of viruses in thin sections of cells grown in culture. Prog. Med. Virol.15:331-379.","journal-title":"Prog. Med. Virol."},{"key":"e_1_3_1_15_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid0703.017311"},{"key":"e_1_3_1_16_2","first-page":"631","volume":"67","year":"1969","unstructured":"Kempe, C. H., F. Dekking, L. Saint Vincent, A. R. Rao, and A. W. Downie. 1969. Conjunctivitis and subclinical infection in smallpox. J. Hyg.67:631-636.","journal-title":"J. Hyg."},{"key":"e_1_3_1_17_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid0504.990411"},{"key":"e_1_3_1_18_2","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.40.3.753-757.2002"},{"key":"e_1_3_1_19_2","doi-asserted-by":"publisher","DOI":"10.3201\/eid0701.010125"},{"key":"e_1_3_1_20_2","doi-asserted-by":"publisher","DOI":"10.1006\/viro.1994.1288"},{"key":"e_1_3_1_21_2","doi-asserted-by":"publisher","DOI":"10.1099\/0022-1317-75-8-1975"},{"key":"e_1_3_1_22_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0891-5520(05)70189-6"},{"key":"e_1_3_1_23_2","first-page":"106","volume":"51","year":"1974","unstructured":"Mitra, A. C., J. K. Sarkar, and M. K. Mukherjee. 1974. Virus content of smallpox scabs. Bull. W. H. O.51:106-107.","journal-title":"Bull. W. H. O."},{"key":"e_1_3_1_24_2","first-page":"279","volume":"40","year":"1969","unstructured":"Noble, J., Jr., and J. A. Rich. 1969. Transmission of smallpox by contact and by aerosol in Macaca irus. Bull. W. H. O.40:279-286.","journal-title":"Bull. W. H. O."},{"key":"e_1_3_1_25_2","doi-asserted-by":"publisher","DOI":"10.3109\/inf.1976.8.issue-3.01"},{"key":"e_1_3_1_26_2","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.33.8.2069-2076.1995"},{"key":"e_1_3_1_27_2","first-page":"13","volume":"55","year":"1967","unstructured":"Sarkar, J. K., and A. C. Mitra. 1967. Virulence of variola virus isolated from smallpox cases of varying severity. Indian J. Med Res.55:13-20.","journal-title":"Indian J. Med Res."},{"key":"e_1_3_1_28_2","first-page":"1800","volume":"62","year":"1974","unstructured":"Sarkar, J. K., A. C. Mitra, and M. K. Mukherjee. 1974. Duration of virus excretion in the throat of asymptomatic household contacts of smallpox patients. Indian J. Med Res.62:1800-1803.","journal-title":"Indian J. Med Res."},{"key":"e_1_3_1_29_2","first-page":"517","volume":"48","year":"1973","unstructured":"Sarkar, J. K., A. C. Mitra, M. K. Mukherjee, S. K. De, and D. G. Muzumdar. 1973. Virus excretion in smallpox. 1. Excretion in the throat, urine, and conjunctiva of patients. Bull. W. H. O.48:517-522.","journal-title":"Bull. W. H. O."},{"key":"e_1_3_1_30_2","first-page":"189","year":"2002","unstructured":"Smith, T. F., M. J. Espy, and A. D. Wold. 2002. Development, implementation, and optimization of LightCycler PCR assays for detection of herpes simplex virus and varicella-zoster virus from clinical specimens, p. 189-200. In U. Reischl, C. Wittwer, and F. Cockerill (ed.), Rapid cycle real-time PCR methods and applications in microbiology food analysis. Springer, New York, N.Y.","journal-title":"Rapid cycle real-time PCR methods and applications in microbiology food analysis"},{"key":"e_1_3_1_31_2","doi-asserted-by":"publisher","DOI":"10.1128\/jcm.13.4.723-725.1981"},{"key":"e_1_3_1_32_2","first-page":"145","volume":"55","year":"1980","unstructured":"World Health Organization. 1980. Declaration of global eradication of smallpox. Wkly. Epidemiol. Rec.55:145-152.","journal-title":"Wkly. Epidemiol. Rec."}],"updated-by":[{"DOI":"10.1128\/jcm.40.11.4405.2002-a","type":"correction","label":"Correction","source":"publisher","updated":{"date-parts":[[2002,11,1]],"date-time":"2002-11-01T00:00:00Z","timestamp":1036108800000}}],"container-title":["Journal of Clinical Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/JCM.40.6.1985-1988.2002","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.asm.org\/doi\/pdf\/10.1128\/JCM.40.6.1985-1988.2002","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,3,5]],"date-time":"2022-03-05T01:49:21Z","timestamp":1646444961000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.asm.org\/doi\/10.1128\/JCM.40.6.1985-1988.2002"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2002,6]]},"references-count":31,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2002,6]]}},"alternative-id":["10.1128\/JCM.40.6.1985-1988.2002"],"URL":"https:\/\/doi.org\/10.1128\/jcm.40.6.1985-1988.2002","relation":{"correction":[{"id-type":"doi","id":"10.1128\/JCM.40.11.4405.2002-a","asserted-by":"object"}]},"ISSN":["0095-1137","1098-660X"],"issn-type":[{"value":"0095-1137","type":"print"},{"value":"1098-660X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2002,6]]},"assertion":[{"value":"2002-03-19","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2002-03-20","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2002-06-01","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}