{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T17:50:49Z","timestamp":1760205049148},"reference-count":35,"publisher":"American Society for Microbiology","issue":"12","license":[{"start":{"date-parts":[[2000,12,1]],"date-time":"2000-12-01T00:00:00Z","timestamp":975628800000},"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":[[2000,12]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n          <jats:p>\n            The combination of preventive vaccination and diagnostic typing of viral isolates from patients with clinical poliomyelitis constitutes our main protective shield against polioviruses. The restriction fragment length polymorphism (RFLP) adaptation of the reverse transcriptase (RT)-PCR methodology has advanced diagnostic genotyping of polioviruses, although further improvements are definitely needed. We report here on an improved RFLP procedure for the genotyping of polioviruses. A highly conserved segment within the 5\u2032 noncoding region of polioviruses was selected for RT-PCR amplification by the UC\n            <jats:sub>53<\/jats:sub>\n            -UG\n            <jats:sub>52<\/jats:sub>\n            primer pair with the hope that it would be most resistant to the inescapable genetic alteration-drift experienced by the other segments of the viral genome. Complete inter- and intratypic genotyping of polioviruses by the present RFLP method was accomplished with a minimum set of four restriction endonucleases (\n            <jats:italic>Hae<\/jats:italic>\n            III,\n            <jats:italic>Dde<\/jats:italic>\n            I,\n            <jats:italic>Nco<\/jats:italic>\n            I, and\n            <jats:italic>Ava<\/jats:italic>\n            I). To compensate for potential genetic drift within the recognition sites of\n            <jats:italic>Hae<\/jats:italic>\n            III,\n            <jats:italic>Dde<\/jats:italic>\n            I, or\n            <jats:italic>Nco<\/jats:italic>\n            I in atypical clinical samples, the RFLP patterns generated with\n            <jats:italic>Hpa<\/jats:italic>\n            II and\n            <jats:italic>Sty<\/jats:italic>\n            I as replacements were analyzed. The specificity of the method was also successfully assessed by RFLP analysis of 55 reference nonpoliovirus enterovirus controls. The concerted implementation of these conditional protocols for diagnostic inter- and intratypic genotyping of polioviruses was evaluated with 21 clinical samples with absolute success.\n          <\/jats:p>","DOI":"10.1128\/jcm.38.12.4337-4342.2000","type":"journal-article","created":{"date-parts":[[2019,12,31]],"date-time":"2019-12-31T17:51:26Z","timestamp":1577814686000},"page":"4337-4342","update-policy":"http:\/\/dx.doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":27,"title":["Improved Genotyping Vaccine and Wild-Type Poliovirus Strains by Restriction Fragment Length Polymorphism Analysis: Clinical Diagnostic Implications"],"prefix":"10.1128","volume":"38","author":[{"given":"Amalia","family":"Georgopoulou","sequence":"first","affiliation":[{"name":"<!--label omitted: 1-->Department of Virology, Hellenic Pasteur Institute, 11521, Athens,1 and"},{"name":"<!--label omitted: 2-->Department of Biology &amp; Genetics, University of Thessaly Medical School, Larisa 41222,2 Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Panayotis","family":"Markoulatos","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->Department of Virology, Hellenic Pasteur Institute, 11521, Athens,1 and"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Niki","family":"Spyrou","sequence":"additional","affiliation":[{"name":"<!--label omitted: 1-->Department of Virology, Hellenic Pasteur Institute, 11521, Athens,1 and"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nicholas C.","family":"Vamvakopoulos","sequence":"additional","affiliation":[{"name":"<!--label omitted: 2-->Department of Biology &amp; Genetics, University of Thessaly Medical School, Larisa 41222,2 Greece"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/0042-6822(91)90434-D"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/168.4.888"},{"key":"e_1_3_2_4_2","first-page":"234","article-title":"Differentiation of types of poliomyelitis viruses","volume":"49","author":"Bodian D.","year":"1949","unstructured":"Bodian D. 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