Identification of Penicillin and other Beta-Lactam Resistance in Streptococcus Pneumoniae by Polymerase Chain Reaction

Ubukata, Kimiko; Muraki, Tomoko; Igarashi, Atsumi; Asahi, Yasuko; Konno, Masatoshi

doi:10.1007/bf02490033

Cited by 52 publications

(68 citation statements)

References 16 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: 88%

“…However, alarmingly high frequencies of penicillin-and macrolide-resistant pneumococci have been reported, especially in several Asian countries, including Japan (4,24,30). The resistance of S. pneumoniae to penicillin has been shown to be closely associated with mosaic mutations in the pbp1a, pbp2b, and pbp2x genes (12,35). Macrolide resistance is generally mediated by two mechanisms: 23S rRNA methylation encoded by the erm(B) gene or macrolide efflux via the mef(A) gene (23,32).…”

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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: 88%

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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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“…The lytA gene encoding autolysin enzyme as well as the pneumolysin gene are specific to S. pneumoniae (26,29); therefore, the lytA gene can differentiate S. pneumoniae from the genotypically similar Streptococcus species (20,35). Similarly, the mip gene of L. pneumophila was selected as the MB sequence because the gene is an encoding macrophage infectivity potentiator and is identified only in L. pneumophila.…”

Section: Patients and Samplesmentioning

confidence: 99%

Simultaneous Detection of Pathogens in Clinical Samples from Patients with Community-Acquired Pneumonia by Real-Time PCR with Pathogen-Specific Molecular Beacon Probes

et al. 2006

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In this study, real-time PCR with pathogen-specific molecular beacons (MB) and primers was evaluated for prediction of community-acquired pneumonia (CAP) causative agents, detecting six main CAP agents, Streptococcus pneumoniae, Haemophilus influenzae, Mycoplasma pneumoniae, Chlamydophila pneumoniae, Legionella pneumophila, and Streptococcus pyogenes, simultaneously. The PCR assay was evaluated for fresh clinical specimens from infants and children (n ‫؍‬ 389) and from adults (n ‫؍‬ 40). The MB probes and primers are both pathogen specific, namely, the lytA gene for S. pneumoniae, the mip gene for L. pneumophila, and 16S rRNA genes for the remaining four organisms. DNA extraction of clinical specimens was performed with a commercially available EXTRAGEN II kit, and amplification was performed with Stratagene Mx3000P. The limit of detection for these pathogens ranged from 2 copies to 18 copies. The whole process from DNA extraction to the analysis was finished in less than 2 h. The obtained sensitivity and specificity of this real-time PCR study relative to those of conventional cultures were as follows: 96.2% and 93.2% for S. pneumoniae, 95.8% and 95.4% for H. influenzae, 100% and 100% for S. pyogenes, and 100% and 95.4% for M. pneumoniae, respectively. The sensitivity and specificity for M. pneumoniae relative to those of a serologic assay were 90.2% and 97.9%, respectively. In six clinical samples of C. pneumoniae, the real-time PCR gave positive predictable values, and in those cases, elevation of the titer value was also observed. In conclusion, we demonstrated that a real-time PCR assay with pathogen-specific MB is useful in identifying CAP causative agents rapidly and in examining the clinical course of empirical chemotherapy in a timely manner, supporting conventional culture methods.Community-acquired pneumonia (CAP) is still a major threat to individuals, especially children and compromised hosts, such as senior citizens and people with underlying chronic diseases. The main causative pathogens in CAP are Streptococcus pneumoniae (30% to 35%), Haemophilus influenzae (4% to 20%), and Mycoplasma pneumoniae (15% to 20%), and the rates change according to age and the underlying disease (5, 7, 16). Recently, Chlamydophila pneumoniae and Legionella pneumophila have frequently been identified as causative pathogens of CAP (9, 30).In clinical practice, empirical chemotherapy with broadspectrum antibiotics must be started based on clinical symptoms, radiograph findings, and clinical examinations, such as those for white blood cell (WBC) counts and C-reactive protein (CRP), considering the severity of the symptoms.On the other hand, the increase of resistant bacteria, including CAP pathogens, is a worldwide health problem (13,22,34). One effective measure to prevent new emergence of resistant bacteria is to use only the sufficient quantity of antibiotics to eliminate the target pathogens. For the measure, the rapid and precise determination of causative pathogens is critical, and a number of studies have been repo...

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“…In Japan, prevalence of PEN-resistant S. pneumoniae (PRSP) among clinical isolates from acute otitis media and respiratory tract infections (RTIs) has been increasing rapidly, especially in younger children (40,41). In parallel with overall increases in incidence of PRSP, meningitis caused by PRSP is being reported increasingly throughout Japan (4).…”

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Antibiotic Susceptibility in Relation to Penicillin-Binding Protein Genes and Serotype Distribution of Streptococcus pneumoniae Strains Responsible for Meningitis in Japan, 1999 to 2002

Ubukata

Chiba

Hasegawa

et al. 2004

Antimicrob Agents Chemother

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The antibiotic susceptibilities, genotypes of penicillin (PEN)-binding protein genes (pbp), and serotype distributions of Streptococcus pneumoniae isolates from meningitis patients were investigated by a nationwide surveillance group in Japan between 1999 and 2002. We analyzed 146 isolates from children (<17 years old) and 73 from adults (>18 years old). Isolates with or without abnormal pbp1a, pbp2x, or pbp2b genes identified by PCR were classified into six genotype patterns and 90% MIC (MIC 90 ) values for PEN: (i) strains with three normal genes (17.2% of isolates; MIC 90 , 0.031 g/ml); (ii) strains with abnormal pbp2x (22.1%, 0.063 g/ml); (iii) strains with abnormal pbp2b (1.0%, 0.125 g/ml); (iv) strains with abnormal pbp2x and pbp2b (7.4%, 0.25 g/ml); (v) strains with abnormal pbp1a and pbp2x (12.7%, 0.25 g/ml); and (vi) strains with three abnormal PBP genes (39.7%, 4 g/ml), which are termed genotypic PEN-resistant S. pneumoniae (gPRSP). Panipenem, a carbapenem, showed an excellent MIC 90 (0.125 g/ml) against gPRSP, followed by meropenem and vancomycin (0.5 g/ml), cefotaxime and ceftriaxone (1 g/ml), and ampicillin (4 g/ml). Strains of gPRSP were significantly more prevalent in children (45.2%) than in adults (27.4%). The most frequent serotypes were 6B, 19F, 23F, 6A, and 14 in children and 23F, 22, 3, 10, 6B, and 19F in adults. Serotypes 6B, 6A, 19F, 23F, and 14 predominated among gPRSP. In children, 7-and 11-valent pneumococcal conjugate vaccines would cover 76.2 and 81.3% of isolates, respectively, although coverage would be lower in adults (43.9 and 56.0%, respectively). These findings suggest the need for early introduction of pneumococcal conjugate vaccines and continuous bacteriological surveillance for meningitis.Streptococcus pneumoniae is a common etiologic agent of serious invasive infections, with high morbidity and mortality in children and adults, such as meningitis, septicemia, and pneumonia (23, 29). The evolution of strains of S. pneumoniae resistant to penicillin G (PEN) and broad-spectrum cephalosporin antibiotics has created difficulties worldwide in selecting an appropriate chemotherapeutic agent (2).Surveillance studies of antibiotic susceptibility, serotype distribution of causative strains, and mortality rate in meningitis have been carried out nationwide in many countries (8,11,15,20,21,28,33,36,39,43).In Japan, prevalence of PEN-resistant S. pneumoniae (PRSP) among clinical isolates from acute otitis media and respiratory tract infections (RTIs) has been increasing rapidly, especially in younger children (40,41). In parallel with overall increases in incidence of PRSP, meningitis caused by PRSP is being reported increasingly throughout Japan (4). As investigators in that country, we felt that nationwide surveillance had become crucial given increasing isolation of resistant strains that could cause meningitis, including PRSP, as well as ampicillin (AMP)-and cephalosporin-resistant strains of Haemophilus influenzae type b, the most frequent etiologic agent of bacterial meningitis. I...

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Identification of Penicillin and other Beta-Lactam Resistance in Streptococcus Pneumoniae by Polymerase Chain Reaction

Cited by 52 publications

References 16 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

Simultaneous Detection of Pathogens in Clinical Samples from Patients with Community-Acquired Pneumonia by Real-Time PCR with Pathogen-Specific Molecular Beacon Probes

Antibiotic Susceptibility in Relation to Penicillin-Binding Protein Genes and Serotype Distribution of Streptococcus pneumoniae Strains Responsible for Meningitis in Japan, 1999 to 2002

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