Pyrosequencing Using the Single-Nucleotide Polymorphism Protocol for Rapid Determination of TEM- and SHV-Type Extended-Spectrum β-Lactamases in Clinical Isolates and Identification of the Novel β-Lactamase Genes bla SHV-48 , bla SHV-105 , and bla TEM-155
| Autor: | Peter Petersen, Patricia A. Bradford, C. Hal Jones, Margareta Tuckman, Alexey Ruzin, Melissa A. Visalli |
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| Rok vydání: | 2009 |
| Předmět: |
Pharmacology
Genetics medicine.drug_class medicine.medical_treatment Cephalosporin Ceftazidime Aztreonam biochemical phenomena metabolism and nutrition Biology bacterial infections and mycoses medicine.disease_cause chemistry.chemical_compound Infectious Diseases Plasmid chemistry Genotype polycyclic compounds medicine Beta-lactamase bacteria Pyrosequencing Pharmacology (medical) Escherichia coli medicine.drug |
| Zdroj: | Antimicrobial Agents and Chemotherapy. 53:977-986 |
| ISSN: | 1098-6596 0066-4804 |
| Popis: | TEM- and SHV-type extended-spectrum β-lactamases (ESBLs) are the most common ESBLs found in the United States and are prevalent throughout the world. Amino acid substitutions at a number of positions in TEM-1 lead to the ESBL phenotype, although substitutions at residues 104 (E to K), 164 (R to S or H), 238 (G to S), and 240 (E to K) appear to be particularly important in modifying the spectrum of activity of the enzyme. The SHV-1-derived ESBLs are a less diverse collection of enzymes; however, the majority of amino acid substitutions resulting in an ESBL mirror those seen in the TEM-1-derived enzymes. Pyrosequencing by use of the single-nucleotide polymorphism (SNP) protocol was applied to provide sequence data at positions critical for the ESBL phenotype spanning the bla TEM and bla SHV genes. Three novel β-lactamases are described: the ESBLs TEM-155 (Q39K, R164S, E240K) and SHV-105 (I8F, R43S, G156D, G238S, E240K) and a non-ESBL, SHV-48 (V119I). The ceftazidime, ceftriaxone, and aztreonam MICs for an Escherichia coli isolate expressing bla SHV-105 were >128, 128, and >128 μg/ml, respectively. Likewise, the ceftazidime, ceftriaxone, and aztreonam MICs for an E. coli isolate expressing bla TEM-155 were >128, 64, and > 128 μg/ml, respectively. Pyrosequence analysis determined the true identity of the β-lactamase on plasmid R1010 to be SHV-11 rather than SHV-1, as previously reported. Pyrosequencing is a real-time sequencing-by-synthesis approach that was applied to SNP detection for TEM- and SHV-type ESBL identification and represents a robust tool for rapid sequence determination that may have a place in the clinical setting. |
| Databáze: | OpenAIRE |
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