{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,14]],"date-time":"2026-05-14T23:24:00Z","timestamp":1778801040245,"version":"3.51.4"},"reference-count":32,"publisher":"SAGE Publications","issue":"2","license":[{"start":{"date-parts":[[2012,4,1]],"date-time":"2012-04-01T00:00:00Z","timestamp":1333238400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/journals.sagepub.com\/page\/policies\/text-and-data-mining-license"}],"content-domain":{"domain":["journals.sagepub.com"],"crossmark-restriction":true},"short-container-title":["Microbial Drug Resistance"],"published-print":{"date-parts":[[2012,4]]},"abstract":"<jats:p>\n                    We sought to identify genotypic resistance classes by real-time PCR in 300\n                    <jats:italic toggle=\"yes\">Streptococcus pneumoniae<\/jats:italic>\n                    isolates from invasive pneumococcal diseases. Primers and molecular beacon probes were designed for the\n                    <jats:italic toggle=\"yes\">lytA<\/jats:italic>\n                    gene, 3\n                    <jats:italic toggle=\"yes\">pbp<\/jats:italic>\n                    genes, and the\n                    <jats:italic toggle=\"yes\">mef<\/jats:italic>\n                    A\/\n                    <jats:italic toggle=\"yes\">erm<\/jats:italic>\n                    B genes. Targeted sequences of\n                    <jats:italic toggle=\"yes\">pbp1a<\/jats:italic>\n                    ,\n                    <jats:italic toggle=\"yes\">pbp2x<\/jats:italic>\n                    , and\n                    <jats:italic toggle=\"yes\">pbp2b<\/jats:italic>\n                    genes in susceptible strain R6 corresponded to those of penicillin G-nonsusceptible strains, including sites within or adjacent to conserved amino acid motifs. If amplification did not occur, the corresponding penicillin-binding protein (PBP) was considered to possess amino acid substitution(s) affecting minimal inhibitory concentrations (MICs) of \u03b2-lactam antibiotics. Real-time PCR required 90\u2009min or less. Strains were assigned to six genotypic classes: Genotypic penicillin-susceptible\n                    <jats:italic toggle=\"yes\">S. pneumoniae<\/jats:italic>\n                    (gPSSP) with 3 normal genes (22.3%); genotypic penicillin-intermediate\n                    <jats:italic toggle=\"yes\">S. pneumoniae<\/jats:italic>\n                    (gPISP) (\n                    <jats:italic toggle=\"yes\">pbp2x<\/jats:italic>\n                    ) with an abnormal\n                    <jats:italic toggle=\"yes\">pbp2x<\/jats:italic>\n                    gene (25.3%); gPISP (\n                    <jats:italic toggle=\"yes\">pbp2b<\/jats:italic>\n                    ) with an abnormal\n                    <jats:italic toggle=\"yes\">pbp2b<\/jats:italic>\n                    gene (7.3%); gPISP (\n                    <jats:italic toggle=\"yes\">pbp1a<\/jats:italic>\n                    +\n                    <jats:italic toggle=\"yes\">2x<\/jats:italic>\n                    ) with abnormal\n                    <jats:italic toggle=\"yes\">pbp1a<\/jats:italic>\n                    +\n                    <jats:italic toggle=\"yes\">2x<\/jats:italic>\n                    genes (11.3%); gPISP (\n                    <jats:italic toggle=\"yes\">pbp2x<\/jats:italic>\n                    +\n                    <jats:italic toggle=\"yes\">2b<\/jats:italic>\n                    ) with abnormal\n                    <jats:italic toggle=\"yes\">pbp2x<\/jats:italic>\n                    +\n                    <jats:italic toggle=\"yes\">2b<\/jats:italic>\n                    genes (4.7%); or genotypic penicillin-resistant\n                    <jats:italic toggle=\"yes\">S. pneumoniae<\/jats:italic>\n                    (gPRSP) with 3 abnormal PBP genes (29.0%). Sensitivity and specificity of real-time PCR compared with those of conventional PCR were high, 73.7\u2013100% and 97.7\u2013100%, respectively. As for relationships between genotype and \u03b2-lactam MICs, 90% of MICs for every resistance class were distributed within three serial dilutions for almost all antibiotics. MICs of each \u03b2-lactam antibiotic were estimated with high probability from genotypic patterns. In conclusion, determination of genotypic classes of\n                    <jats:italic toggle=\"yes\">S. pneumoniae<\/jats:italic>\n                    using rapid real-time PCR is useful in selecting effective therapeutic agents for patients with pneumococcal infection.\n                  <\/jats:p>","DOI":"10.1089\/mdr.2011.0102","type":"journal-article","created":{"date-parts":[[2011,10,26]],"date-time":"2011-10-26T10:46:24Z","timestamp":1319625984000},"page":"149-156","update-policy":"https:\/\/doi.org\/10.1177\/sage-journals-update-policy","source":"Crossref","is-referenced-by-count":18,"title":["Application of the Real-Time PCR Method for Genotypic Identification of \u03b2-Lactam Resistance in Isolates from Invasive Pneumococcal Diseases"],"prefix":"10.1177","volume":"18","author":[{"given":"Naoko","family":"Chiba","sequence":"first","affiliation":[{"name":"Kitasato University","place":["Japan"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Miyuki","family":"Morozumi","sequence":"additional","affiliation":[{"name":"Kitasato University","place":["Japan"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kimiko","family":"Ubukata","sequence":"additional","affiliation":[{"name":"Kitasato University","place":["Japan"]}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"179","published-online":{"date-parts":[[2012,4]]},"reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1093\/clinids\/15.1.77"},{"key":"e_1_3_2_3_2","doi-asserted-by":"crossref","first-page":"1252","DOI":"10.1128\/AAC.43.5.1252","article-title":"Diversity of substitutions within or adjacent to conserved amino acid motifs of penicillin-binding protein 2X in cephalosporin-resistant Streptococcus pneumoniae isolates","volume":"43","author":"Asahi Y.","year":"1999","unstructured":"Asahi Y., , Takeuchi Y., , Ubukata K. 1999. Diversity of substitutions within or adjacent to conserved amino acid motifs of penicillin-binding protein 2X in cephalosporin-resistant Streptococcus pneumoniae isolates. Antimicrob. Agents Chemother., 43:1252\u20131255.","journal-title":"Antimicrob. Agents Chemother."},{"key":"e_1_3_2_4_2","doi-asserted-by":"crossref","first-page":"2267","DOI":"10.1128\/AAC.42.9.2267","article-title":"Association of a thr-371 substitution in a conserved amino acid motif of penicillin-binding protein 1A with penicillin resistance of Streptococcus pneumoniae","volume":"42","author":"Asahi Y.","year":"1998","unstructured":"Asahi Y., , Ubukata K. 1998. Association of a thr-371 substitution in a conserved amino acid motif of penicillin-binding protein 1A with penicillin resistance of Streptococcus pneumoniae. Antimicrob. Agents Chemother., 42:2267\u20132273.","journal-title":"Antimicrob. Agents Chemother."},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1097\/INF.0b013e318124a494"},{"key":"e_1_3_2_6_2","first-page":"64","article-title":"Pneumococcal polysaccharide vaccine","volume":"38","author":"Centers for Disease Control (CDC)","year":"1989","unstructured":"Centers for Disease Control (CDC). 1989. Pneumococcal polysaccharide vaccine. MMWR Morb. Mortal Wkly. Rep., 38:64\u20136873\u201376.","journal-title":"MMWR Morb. Mortal Wkly. Rep."},{"key":"e_1_3_2_7_2","first-page":"893","article-title":"Direct and indirect effects of routine vaccination of children with 7-valent pneumococcal conjugate vaccine on incidence of invasive pneumococcal disease\u2014United States, 1998\u20132003","author":"Centers for Disease Control, Prevention (CDC)","year":"2005","unstructured":"Centers for Disease Control, Prevention (CDC). 2005. Direct and indirect effects of routine vaccination of children with 7-valent pneumococcal conjugate vaccine on incidence of invasive pneumococcal disease\u2014United States, 1998\u20132003. MMWR Morb. Mortal Wkly. Rep., 893\u2013897.","journal-title":"MMWR Morb. Mortal Wkly. Rep."},{"key":"e_1_3_2_8_2","first-page":"144","article-title":"Invasive pneumococcal disease in children 5 years after conjugate vaccine introduction\u2014eight states, 1998\u20132005","volume":"57","author":"Centers for Disease Control, Prevention (CDC)","year":"2008","unstructured":"Centers for Disease Control, Prevention (CDC). 2008. Invasive pneumococcal disease in children 5 years after conjugate vaccine introduction\u2014eight states, 1998\u20132005. MMWR Morb. Mortal Wkly. Rep., 57:144\u2013148.","journal-title":"MMWR Morb. Mortal Wkly. Rep."},{"key":"e_1_3_2_9_2","doi-asserted-by":"crossref","first-page":"61","DOI":"10.1017\/S0950268809990239","article-title":"Serotype and antibiotic resistance of isolates from patients with invasive pneumococcal disease in Japan","volume":"138","author":"Chiba N.","year":"2010","unstructured":"Chiba N., , Morozumi M., , Sunaoshi K., , Takahashi S., , Takano M., , Komori T., , Sunakawa K., , Ubukata K. 2010. Serotype and antibiotic resistance of isolates from patients with invasive pneumococcal disease in Japan. Epidemiol. Infect., 138:61\u201368.","journal-title":"Epidemiol. Infect."},{"key":"e_1_3_2_10_2","doi-asserted-by":"crossref","first-page":"8842","DOI":"10.1073\/pnas.86.22.8842","article-title":"Horizontal transfer of penicillin-binding protein genes in penicillin-resistant clinical isolates of Streptococcus pneumoniae","volume":"86","author":"Dowson C.G.","year":"1989","unstructured":"Dowson C.G., , Hutchison A., , Brannigan J.A, , George R.C., , Hansman D., , Linares J., , Tomasz A., , Smith J.M., , Spratt B.G. 1989. Horizontal transfer of penicillin-binding protein genes in penicillin-resistant clinical isolates of Streptococcus pneumoniae. Proc. Natl. Acad. Sci. USA, 86:8842\u20138846.","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"e_1_3_2_11_2","doi-asserted-by":"crossref","first-page":"7518","DOI":"10.1093\/nar\/17.18.7518","article-title":"Nucleotide sequence of the penicillin-binding protein 2B gene of Streptococcus pneumoniae strain R6","volume":"17","author":"Dowson C.G.","year":"1989","unstructured":"Dowson C.G., , Hutchison A., , Spratt B.G. 1989. Nucleotide sequence of the penicillin-binding protein 2B gene of Streptococcus pneumoniae strain R6. Nucleic Acids Res., 17:7518.","journal-title":"Nucleic Acids Res."},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1097\/01.inf.0000253059.84602.c3"},{"key":"e_1_3_2_13_2","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1016\/0378-1119(86)90215-5","article-title":"Nucleotide sequence and expression of the pneumococcal autolysin gene from its own promoter in Escherichia coli","volume":"43","author":"Garcia P.","year":"1986","unstructured":"Garcia P., , Garcia J.L., , Garcia E., , Lopez R. 1986. Nucleotide sequence and expression of the pneumococcal autolysin gene from its own promoter in Escherichia coli. Gene, 43:265\u2013272.","journal-title":"Gene"},{"key":"e_1_3_2_14_2","doi-asserted-by":"crossref","first-page":"829","DOI":"10.1128\/AAC.40.4.829","article-title":"Penicillin-binding proteins 2b and 2x of Streptococcus pneumoniae are primary resistance determinants for different classes of beta-lactam antibiotics","volume":"40","author":"Grebe T.","year":"1996","unstructured":"Grebe T., , Hakenbeck R. 1996. Penicillin-binding proteins 2b and 2x of Streptococcus pneumoniae are primary resistance determinants for different classes of beta-lactam antibiotics. Antimicrob. Agents Chemother., 40:829\u2013834.","journal-title":"Antimicrob. Agents Chemother."},{"key":"e_1_3_2_15_2","doi-asserted-by":"crossref","first-page":"364","DOI":"10.1128\/AAC.17.3.364","article-title":"Multiple changes of penicillin-binding proteins in penicillin-resistant clinical isolates of Streptococcus pneumoniae","volume":"17","author":"Hakenbeck R.","year":"1980","unstructured":"Hakenbeck R., , Tarpay M., , Tomasz A. 1980. Multiple changes of penicillin-binding proteins in penicillin-resistant clinical isolates of Streptococcus pneumoniae. Antimicrob. Agents Chemother., 17:364\u2013371.","journal-title":"Antimicrob. Agents Chemother."},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1128\/CMR.3.2.171"},{"key":"e_1_3_2_17_2","doi-asserted-by":"crossref","first-page":"1337","DOI":"10.1111\/j.1365-2958.1989.tb00115.x","article-title":"Nucleotide sequences of the pbpX genes encoding the penicillin-binding proteins 2x from Streptococcus pneumoniae R6 and a cefotaxime-resistant mutant, C506","volume":"3","author":"Laible G.","year":"1989","unstructured":"Laible G., , Hakenbeck R., , Sicard M.A., , Joris B., , Ghuysen J.M. 1989. Nucleotide sequences of the pbpX genes encoding the penicillin-binding proteins 2x from Streptococcus pneumoniae R6 and a cefotaxime-resistant mutant, C506. Mol. Microbiol., 3:1337\u20131348.","journal-title":"Mol. Microbiol."},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1001\/jama.294.16.2043"},{"key":"e_1_3_2_19_2","doi-asserted-by":"crossref","first-page":"4517","DOI":"10.1128\/jb.174.13.4517-4523.1992","article-title":"Nucleotide sequences of genes encoding penicillin-binding proteins from Streptococcus pneumoniae and Streptococcus oralis with high homology to Escherichia coli penicillin-binding proteins 1a and 1b","volume":"174","author":"Martin C.","year":"1992","unstructured":"Martin C., , Briese T., , Hakenbeck R. 1992. Nucleotide sequences of genes encoding penicillin-binding proteins from Streptococcus pneumoniae and Streptococcus oralis with high homology to Escherichia coli penicillin-binding proteins 1a and 1b. J. Bacteriol., 174:4517\u20134523.","journal-title":"J. Bacteriol."},{"key":"e_1_3_2_20_2","doi-asserted-by":"crossref","first-page":"915","DOI":"10.1093\/jac\/48.6.915","article-title":"Evaluation of PCR primers to screen for Streptococcus pneumoniae isolates and beta-lactam resistance, and to detect common macrolide resistance determinants","volume":"48","author":"Nagai K.","year":"2001","unstructured":"Nagai K., , Shibasaki Y., , Hasegawa K., , Davies T.A., , Jacobs M.R., , Ubukata K., , Appelbaum P.C. 2001. Evaluation of PCR primers to screen for Streptococcus pneumoniae isolates and beta-lactam resistance, and to detect common macrolide resistance determinants. J Antimicrob Chemother, 48:915\u20138.","journal-title":"J Antimicrob Chemother"},{"key":"e_1_3_2_21_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(09)61204-6"},{"key":"e_1_3_2_22_2","doi-asserted-by":"crossref","first-page":"468","DOI":"10.1097\/INF.0b013e31803df9ca","article-title":"Emergence of 19A as virulent and multidrug resistant Pneumococcus in Massachusetts following universal immunization of infants with pneumococcal conjugate vaccine","volume":"26","author":"Pelton S.I.","year":"2007","unstructured":"Pelton S.I., , Huot H., , Finkelstein J.A., , Bishop C.J., , Hsu K.K., , Kellenberg J., , Huang S.S., , Goldstein R., , Hanage W.P. 2007. Emergence of 19A as virulent and multidrug resistant Pneumococcus in Massachusetts following universal immunization of infants with pneumococcal conjugate vaccine. Pediatr. Infect. Dis. J, 26:468\u2013472.","journal-title":"Pediatr. Infect. Dis. J"},{"key":"e_1_3_2_23_2","doi-asserted-by":"publisher","DOI":"10.1001\/jama.295.14.1668"},{"key":"e_1_3_2_24_2","doi-asserted-by":"crossref","first-page":"152","DOI":"10.1128\/AAC.40.1.152","article-title":"In vitro selection of one-step mutants of Streptococcus pneumoniae resistant to different oral beta-lactam antibiotics is associated with alterations of PBP2x","volume":"40","author":"Sifaoui F.","year":"1996","unstructured":"Sifaoui F., , Kitzis M.D., , Gutmann L. 1996. In vitro selection of one-step mutants of Streptococcus pneumoniae resistant to different oral beta-lactam antibiotics is associated with alterations of PBP2x. Antimicrob. Agents Chemother., 40:152\u2013156.","journal-title":"Antimicrob. Agents Chemother."},{"key":"e_1_3_2_25_2","doi-asserted-by":"crossref","first-page":"1329","DOI":"10.1128\/AAC.42.6.1329","article-title":"Alterations in PBP 1A essential-for high-level penicillin resistance in Streptococcus pneumoniae","volume":"42","author":"Smith A.M.","year":"1998","unstructured":"Smith A.M., , Klugman K.P. 1998. Alterations in PBP 1A essential-for high-level penicillin resistance in Streptococcus pneumoniae. Antimicrob. Agents Chemother., 42:1329\u20131333.","journal-title":"Antimicrob. Agents Chemother."},{"key":"e_1_3_2_26_2","doi-asserted-by":"crossref","first-page":"2251","DOI":"10.1128\/AAC.41.10.2251","article-title":"mefE is necessary for the erythromycin-resistant M phenotype in Streptococcus pneumoniae","volume":"41","author":"Tait-Kamradt A.","year":"1997","unstructured":"Tait-Kamradt A., , Clancy J., , Cronan M., , Dib-Hajj F., , Wondrack L., , Yuan W., , Sutcliffe J. 1997. mefE is necessary for the erythromycin-resistant M phenotype in Streptococcus pneumoniae. Antimicrob. Agents Chemother., 41:2251\u20132255.","journal-title":"Antimicrob. Agents Chemother."},{"key":"e_1_3_2_27_2","doi-asserted-by":"publisher","DOI":"10.1097\/INF.0b013e3181c2a229"},{"key":"e_1_3_2_28_2","doi-asserted-by":"crossref","first-page":"3660","DOI":"10.1093\/nar\/18.12.3660","article-title":"Nucleotide sequence of the erythromycin resistance gene of the conjugative transposon Tn1545","volume":"18","author":"Trieu-Cuot P.","year":"1990","unstructured":"Trieu-Cuot P., , Poyart-Salmeron C., , Carlier C., , Courvalin P. 1990. Nucleotide sequence of the erythromycin resistance gene of the conjugative transposon Tn1545. Nucleic Acids Res., 18:3660.","journal-title":"Nucleic Acids Res."},{"key":"e_1_3_2_29_2","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1007\/BF02350645","article-title":"Incidence of penicillin-resistant Streptococcus pneumoniae in Japan, 1993\u20131995","volume":"1","author":"Ubukata K.","year":"1996","unstructured":"Ubukata K., , Asahi Y., , Okuzumi K., , Konno M. 1996. Incidence of penicillin-resistant Streptococcus pneumoniae in Japan, 1993\u20131995. J. Infect. Chemother., 1:177\u2013184.","journal-title":"J. Infect. Chemother."},{"key":"e_1_3_2_30_2","doi-asserted-by":"crossref","first-page":"592","DOI":"10.1128\/jcm.34.3.592-596.1996","article-title":"Combinational detection of autolysin and penicillin-binding protein 2B genes of Streptococcus pneumoniae by PCR","volume":"34","author":"Ubukata K.","year":"1996","unstructured":"Ubukata K., , Asahi Y., , Yamane A., , Konno M. 1996. Combinational detection of autolysin and penicillin-binding protein 2B genes of Streptococcus pneumoniae by PCR. J. Clin. Microbiol., 34:592\u2013596.","journal-title":"J. Clin. Microbiol."},{"key":"e_1_3_2_31_2","doi-asserted-by":"crossref","first-page":"1488","DOI":"10.1128\/AAC.48.5.1488-1494.2004","article-title":"Antibiotic susceptibility in relation to penicillin-binding protein genes and serotype distribution of Streptococcus pneumoniae strains responsible for meningitis in Japan, 1999 to 2002","volume":"48","author":"Ubukata K.","year":"2004","unstructured":"Ubukata K., , Chiba N., , Hasegawa K., , Kobayashi R., , Iwata S., , Sunakawa K. 2004. Antibiotic susceptibility in relation to penicillin-binding protein genes and serotype distribution of Streptococcus pneumoniae strains responsible for meningitis in Japan, 1999 to 2002. Antimicrob. Agents Chemother., 48:1488\u20131494.","journal-title":"Antimicrob. Agents Chemother."},{"key":"e_1_3_2_32_2","doi-asserted-by":"crossref","first-page":"190","DOI":"10.1007\/BF02490033","article-title":"Identification of penicillin and other beta-lactam resistance in Streptococcus pneumoniae by polymerase chain reaction","volume":"3","author":"Ubukata K.","year":"1997","unstructured":"Ubukata K., , Muraki T., , Igarashi A., , Asahi Y., , Konno M. 1997. Identification of penicillin and other beta-lactam resistance in Streptococcus pneumoniae by polymerase chain reaction. J. Infect. Chemother., 3:190\u2013197.","journal-title":"J. Infect. Chemother."},{"key":"e_1_3_2_33_2","volume-title":"State of the art of new vaccines: Research and development","author":"World Health Organization (WHO)","year":"2005","unstructured":"World Health Organization (WHO). 2005. State of the art of new vaccines: Research and development. WHO: Geneva, Switzerland."}],"container-title":["Microbial Drug Resistance"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.sagepub.com\/doi\/full-xml\/10.1089\/mdr.2011.0102","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.sagepub.com\/doi\/pdf\/10.1089\/mdr.2011.0102","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/journals.sagepub.com\/doi\/pdf\/10.1089\/mdr.2011.0102","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,5,1]],"date-time":"2026-05-01T20:26:38Z","timestamp":1777667198000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.sagepub.com\/doi\/full\/10.1089\/mdr.2011.0102"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2012,4]]},"references-count":32,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2012,4]]}},"alternative-id":["10.1089\/mdr.2011.0102"],"URL":"https:\/\/doi.org\/10.1089\/mdr.2011.0102","relation":{},"ISSN":["1076-6294","1931-8448"],"issn-type":[{"value":"1076-6294","type":"print"},{"value":"1931-8448","type":"electronic"}],"subject":[],"published":{"date-parts":[[2012,4]]}}}