Application of a multiplex pbp2b and pbp2x PCR for prediction of penicillin resistance in Streptococcus pneumoniae

Ho, Pak Leung; Wong, River Chun-Wai; Chow, Frankie K.H.; Cheung, May Y M; Wong, S.P.; Yam, Wing Cheong; Que, Tak Lun

doi:10.1093/jac/dkh216

Cited by 3 publications

(4 citation statements)

References 8 publications

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“…However, positive results were not obtained from the mitis group of streptococci (including our collected clinical strains of 17 S. mitis and 12 S. oralis) that were tested for cross-reactivity in the present study. Compared to past reports (13,14,21,34,35), we obtained lower pbp2b detection rates in PCG-intermediate S. pneumoniae strains. This may have been due to a difference in the regions targeted by the pbp2b primers.…”

Section: Nonecontrasting

confidence: 84%

“…The results of these studies suggest that clinical infection correlates with increased pneumococcal load, and these studies also mentioned the capacity of quantitative real-time PCR to distinguish between colonization and true infection (15,36). Many investigators have evaluated the accuracy of multiplex PCR methods, which are used in the screening of S. pneumoniae strains demonstrating penicillin and macrolide resistance (13,14,21,33,34). Most of these assays, however, require separate tubes, are only able to detect two or three gene fragments, and have not been evaluated for clinical respiratory tract samples.…”

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confidence: 99%

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Rapid Identification of Penicillin and Macrolide Resistance Genes and Simultaneous Quantification of Streptococcus pneumoniae in Purulent Sputum Samples by Use of a Novel Real-Time Multiplex PCR Assay

Fukushima¹,

Yanagihara²,

Hirakata

et al. 2008

J Clin Microbiol

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Section: Nonecontrasting

confidence: 84%

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confidence: 99%

Rapid Identification of Penicillin and Macrolide Resistance Genes and Simultaneous Quantification of Streptococcus pneumoniae in Purulent Sputum Samples by Use of a Novel Real-Time Multiplex PCR Assay

Fukushima¹,

Yanagihara²,

Hirakata

et al. 2008

J Clin Microbiol

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“…Therefore, a complete understanding of the resistance mechanism would require the genome-wide comparison of many strains with different or similar resistance profiles, followed by experimental testing through gene transfers. Meanwhile, an imperfect sequence-based method able to rapidly qualify an isolate in the categories susceptible, reduced susceptibility, or likely to be highly resistant (6,8,(14)(15)(16)30) may already be developed and provide some benefit in clinical settings.…”

Section: Discussionmentioning

confidence: 99%

Identical Penicillin-Binding Domains in Penicillin-Binding Proteins of Streptococcus pneumoniae Clinical Isolates with Different Levels of β-Lactam Resistance

Chesnel

Carapito

Croizé

et al. 2005

Antimicrob Agents Chemother

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We have sequenced the penicillin-binding domains of the complete repertoire of penicillin-binding proteins and MurM from 22 clinical isolates of Streptococcus pneumoniae that span a wide range of ␤-lactam resistance levels. Evidence of mosaicism was found in the genes encoding PBP 1a, PBP 2b, PBP 2x, MurM, and, possibly, PBP 2a. Five isolates were found to have identical PBP and MurM sequences, even though the MICs for penicillin G ranged from 0.25 to 2.0 mg/liter. When the sequences encoding PBP 1a, PBP 2b, and PBP 2x from one of these isolates were used to transform laboratory strain R6, the resulting strain had a resistance level higher than that of the less resistant isolates carrying that PBP set but lower than that of the most resistant isolates carrying that PBP set. This result demonstrates that if the R6 strain is arbitrarily defined as the standard genotype, some wild genetic backgrounds can either increase or decrease the PBP-based resistance phenotype.The mounting rates of resistance of Streptococcus pneumoniae to antibiotics is an important concern, as this human pathogen is responsible for serious diseases, such as otitis, pneumonia, meningitis, and bacteremia. In clinical isolates, resistance to ␤-lactams is mostly due to the expression of modified penicillin-binding proteins (PBPs), the targets of the ␤-lactams, with reduced affinities for the antibiotics (3). The low-affinity PBPs are the products of mosaic genes that result from multiple events of homologous recombination with genes from other strains or closely related species (3). The presence of a functional murM gene (also called fibA), which is involved in the synthesis of an alternative physiological substrate for the PBPs, is required for the expression of ␤-lactam resistance (9, 31). Mosaic murM genes can dramatically increase the level of resistance due to low-affinity PBPs (9, 28).S. pneumoniae has five high-molecular-weight PBPs, three of which (PBP 2x, PBP 2b, and PBP 1a) are definitely involved in ␤-lactam resistance (23), and one low-molecular-weight PBP. A laboratory experiment showed that the five high-molecularweight PBPs were modified upon transfer of high-level resistance from Streptococcus mitis to S. pneumoniae (13). Other works have suggested a role for PBP 2a (26,27) and the low-molecular-weight PBP, PBP 3 (17), in ␤-lactam resistance. These findings raised the possibility that previously undetected variability in all the pbp genes may account for the wide range of levels of resistance to ␤-lactams.On the basis of the premise that knowledge of the sequence of MurM and all the PBPs might be sufficient to predict the level of ␤-lactam resistance, we sequenced murM and the region coding the penicillin-binding domain of the six PBPs from 22 isolates of S. pneumoniae. Five isolates were found to have very different MICs for ␤-lactams, even though they had the same six penicillin-binding domains and MurM. This demonstrates that other genes modulate significantly the resistance provided by the low-affinity PBPs. By transferring ...

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“…20 The IMPACT suggested the use of intravenous co-amoxiclav (Augmentin) or sultamicillin (Unasyn) for prophylaxis in closed tube thoracostomy for chest trauma. 13…”

Section: Thoracic Traumamentioning

confidence: 99%

The use of Prophylactic Antibiotics in Trauma

Yuen

2004

Hong Kong Journal of Emergency Medicine

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Application of a multiplex pbp2b and pbp2x PCR for prediction of penicillin resistance in Streptococcus pneumoniae

Cited by 3 publications

References 8 publications

Rapid Identification of Penicillin and Macrolide Resistance Genes and Simultaneous Quantification of Streptococcus pneumoniae in Purulent Sputum Samples by Use of a Novel Real-Time Multiplex PCR Assay

Rapid Identification of Penicillin and Macrolide Resistance Genes and Simultaneous Quantification of Streptococcus pneumoniae in Purulent Sputum Samples by Use of a Novel Real-Time Multiplex PCR Assay

Identical Penicillin-Binding Domains in Penicillin-Binding Proteins of Streptococcus pneumoniae Clinical Isolates with Different Levels of β-Lactam Resistance

The use of Prophylactic Antibiotics in Trauma

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