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Twenty-one<jats:italic>Klebsiella pneumoniae<\/jats:italic>isolates obtained between 1991 and 1995 at the Princess Alexandra Hospital in Brisbane, Australia, were subject to amplification and sequencing of the SHV \u03b2-lactamase-encoding genes. Thirteen strains were phenotypically ESBL positive. Of these, six strains carried the<jats:italic>bla<\/jats:italic><jats:sub>SHV-2a<\/jats:sub>gene and seven strains carried the<jats:italic>bla<\/jats:italic><jats:sub>SHV-12<\/jats:sub>gene. Eight strains were phenotypically ESBL negative. Of these, seven strains carried the non-ESBL<jats:italic>bla<\/jats:italic><jats:sub>SHV-11<\/jats:sub>gene and one strain carried the non-ESBL<jats:italic>bla<\/jats:italic><jats:sub>SHV-1<\/jats:sub>gene. There was complete correspondence between the ESBL phenotype and the presence or absence of an ESBL-encoding gene(s). In addition, it was determined that of the 13 ESBL-positive strains, at least 4 carried copies of a non-ESBL-encoding gene in addition to the<jats:italic>bla<\/jats:italic><jats:sub>SHV-2a<\/jats:sub>or<jats:italic>bla<\/jats:italic><jats:sub>SHV12<\/jats:sub>gene. A minisequencing-based assay was developed to discriminate the different SHV classes. This technique, termed \u201cfirst-nucleotide change,\u201d involves the identification of the base added to a primer in a single-nucleotide extension reaction. The assay targeted polymorphisms at the first bases of codons 238 and 240 and reliably discriminated ESBL-positive strains from ESBL-negative strains and also distinguished strains carrying<jats:italic>bla<\/jats:italic><jats:sub>SHV-2a<\/jats:sub>from strains carrying<jats:italic>bla<\/jats:italic><jats:sub>SHV-12<\/jats:sub>. In addition, this method was used to demonstrate an association between the relative copy numbers of<jats:italic>bla<\/jats:italic><jats:sub>SHV<\/jats:sub>genes in individual strains and the levels of antibiotic resistance.<\/jats:p>","DOI":"10.1128\/aac.46.3.659-664.2002","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T09:58:35Z","timestamp":1027763915000},"page":"659-664","update-policy":"http:\/\/dx.doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":20,"title":["Identification and Minisequencing-Based Discrimination of SHV \u03b2-Lactamases in Nosocomial Infection-Associated<i>Klebsiella pneumoniae<\/i>in Brisbane, Australia"],"prefix":"10.1128","volume":"46","author":[{"given":"Christopher","family":"Howard","sequence":"first","affiliation":[{"name":"Cooperative Research Centre for Diagnostics, Queensland University of Technology, Brisbane, Queensland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Angela","family":"van Daal","sequence":"additional","affiliation":[{"name":"Cooperative Research Centre for Diagnostics, Queensland University of Technology, Brisbane, Queensland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gregory","family":"Kelly","sequence":"additional","affiliation":[{"name":"Cooperative Research Centre for Diagnostics, Queensland University of Technology, Brisbane, Queensland"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jacqueline","family":"Schooneveldt","sequence":"additional","affiliation":[{"name":"Queensland Health Pathology Services, Princess Alexandra Hospital, Brisbane, Queensland, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Graeme","family":"Nimmo","sequence":"additional","affiliation":[{"name":"Queensland Health Pathology Services, Princess Alexandra Hospital, Brisbane, Queensland, Australia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Philip M.","family":"Giffard","sequence":"additional","affiliation":[{"name":"Cooperative Research Centre for Diagnostics, Queensland University of Technology, Brisbane, Queensland"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","first-page":"269","volume":"256","year":"1991","unstructured":"Ambler, R. 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