Comparison of a polymerase chain reaction assay and a conventional microbiologic method for detection of methicillin-resistant Staphylococcus aureus

Tokue, Yutaka; Shoji, Satoru; Satoh, Ken; Watanabe, Akira; Motomiya, Masakichi

doi:10.1128/aac.36.1.6

Cited by 118 publications

(75 citation statements)

References 18 publications

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“…A total of 359 MRSA isolates were collected between 1961 and 1999 from 20 countries. Isolates were confirmed as MRSAs in our laboratory by detecting the presence of the mecA gene with PCR (14). The collection contains members of previously described EMRSA clones, including the Iberian (15), Portuguese͞Brazilian (16), Vienna (17), New York͞Japanlocus to those of the known alleles in the S. aureus MLST database.…”

Section: Methodsmentioning

confidence: 99%

The evolutionary history of methicillin-resistant Staphylococcus aureus (MRSA)

Enright

Robinson²,

Randle³

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

1,433

1,179

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Methicillin-resistant Staphylococcus aureus (MRSA) is a major cause of hospital-acquired infections that are becoming increasingly difficult to combat because of emerging resistance to all current antibiotic classes. The evolutionary origins of MRSA are poorly understood, no rational nomenclature exists, and there is no consensus on the number of major MRSA clones or the relatedness of clones described from different countries. We resolve all of these issues and provide a more thorough and precise analysis of the evolution of MRSA clones than has previously been possible. Using multilocus sequence typing and an algorithm, BURST, we analyzed an international collection of 912 MRSA and methicillinsusceptible S. aureus (MSSA) isolates. We identified 11 major MRSA clones within five groups of related genotypes. The putative ancestral genotype of each group and the most parsimonious patterns of descent of isolates from each ancestor were inferred by using BURST, which, together with analysis of the methicillin resistance genes, established the likely evolutionary origins of each major MRSA clone, the genotype of the original MRSA clone and its MSSA progenitor, and the extent of acquisition and horizontal movement of the methicillin resistance genes. Major MRSA clones have arisen repeatedly from successful epidemic MSSA strains, and isolates with decreased susceptibility to vancomycin, the antibiotic of last resort, are arising from some of these major MRSA clones, highlighting a depressing progression of increasing drug resistance within a small number of ecologically successful S. aureus genotypes.

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Section: Methodsmentioning

confidence: 99%

The evolutionary history of methicillin-resistant Staphylococcus aureus (MRSA)

Enright

Robinson²,

Randle³

et al. 2002

Proc. Natl. Acad. Sci. U.S.A.

1,433

1,179

View full text Add to dashboard Cite

show abstract

“…Among 126 S. aureus, 54 isolates were positive for the mecA gene by PCR, remaining 74 S. aureus were negative (Table I). Schmitz et al, (1997) and Tokue et al, (1992) previously reported that the utility of PCR for the accurate detection of the mecA gene and the possibility of simultaneous identification of S. aureus and detection of mecA gene. During the last decade, many studies have demonstrated the extremely high capacity of PCR for specifically detecting bacteria and genes of interest (Salisbury et al, 1996).…”

Section: Detection Of Meca Gene By Pcrmentioning

confidence: 99%

Performance of CHROM Agar and Oxacillin Resistant Screening Agar Base Media for Detection of Methicillin Resistant Staphylococcus aureus (MRSA) from Chronic Wound

Karthy¹,

Ranjitha²,

Mohankumar³

2009

MAS

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CHROM agar Staphylococcus aureus and Oxacillin Resistant Screening Agar Base (ORSAB) media with oxacillin were evaluated for the screening of Methicillin Resistant Saphylococcus aureus (MRSA). Among 190 samples, totally 126 confirmed Staphylococcus aureus strains were used for screening of MRSA used CHROM agar and ORSAB media were compared with the other MRSA screening media like Baird Park agar (BPA) with ciprofloxacin, Mannitol salt agar (MSA) with oxacillin, Blood agar (BA) with oxacillin and Muller Hinton agar (MHA) with oxacillin. Totally 54 MRSA strains were confirmed using PCR, among that 83% and 92% of the MRSA stains were isolated as mauve colonies on CHROM agar and blue colonies on ORSAB medium in 24 hrs incubation, compare with 64%, 61%, 50%, and 42% of the strains that were isolated on BPA, MSA, BA and MHA respectively. After 48 hrs of incubation 100%, 98%, 77%, 77%, 72% and 68% of the MRSA strains were isolated on CHROM agar, ORSAB, BPA, MSA, BA and MHA respectively. CHROM agar Staphylococcus aureus and ORSAB agar proved to be more sensitivity and specificity than other MRSA selective media. These provide an alternative for the detection of MRSA in clinical laboratories, especially when PCR is unavailable.

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“…The rationale for using ceftizoxime and aztreonam in the selective broth instead of oxacillin and colistin was that earlier studies had shown that both oxacillin and colistin resulted in inhibited or slower growth of MRSA strains (data not shown). Furthermore, ceftizoxime is known to increase the phenotypic level of resistance to methicillin (8,12,13).…”

mentioning

confidence: 99%

Improved Detection of Methicillin-Resistant Staphylococcus aureus Using Phenyl Mannitol Broth Containing Aztreonam and Ceftizoxime

et al. 2001

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We tested a phenyl mannitol broth containing ceftizoxime and aztreonam (PHMB ؉ ) for detection of methicillin-resistant Staphylococcus aureus (MRSA) with reference MRSA strains and, subsequently, with clinical samples (n ‫؍‬ 1,098). All reference MRSA strains induced color change in PHMB ؉ after 24 to 72 h of incubation. In a clinical setting, 40 MRSA strains were detected with PHMB ؉ , compared with only 23 detected with a routine method. Thus, this selective broth significantly (P < 0.001) improved the rate of MRSA detection.Detection of methicillin-resistant Staphylococcus aureus (MRSA) in clinical samples continues to be important, since infections due to MRSA have high morbidity and mortality rates. Moreover, some MRSA strains have the potential to spread rapidly and colonize in other patients. In The Netherlands, therefore, patients who are suspected of being MRSA carriers are isolated until screening cultures are repeatedly negative for MRSA. Methods to detect MRSA in clinical samples ideally should have high sensitivity and a short time to reporting the results. To increase the sensitivity, one can simply take more screening samples on the same day or on consecutive days, but this method is more cumbersome and increase the time to reporting. Another way to increase the sensitivity is to use a broth in addition to agar plates, as was demonstrated previously (1, 3, 11; R. L. Sautter and L. W. Wells, Letter, J. Clin. Microbiol. 28:2380-2381, 1990; M. L. Van Ogtrop, Letter, Antimicrob. Agents Chemother. 39:2169, 1995). To increase the sensitivity of the detection of MRSA from a single sample and to improve laboratory efficiency, we developed a new selective broth containing phenol red, mannitol, aztreonam, and ceftizoxime (PHMB ϩ ). First, we tested the broth with laboratory reference strains. Subsequently, we compared our routine method of direct plating of specimens onto blood agar plates and mannitol salt agars with the new selective broth combined with a blood agar plate.The PHMB ϩ was made by mixing 21 mg of dehydrated phenol red mannitol broth (Becton Dickinson, Le Pont de Claix, France) with 1,000 ml of distilled water, sterilizing the broth for 15 min at 121°C, and letting it cool to room temperature. We then mixed 5 mg of ceftizoxime (Yamanouchi) with 5 ml of distilled water and added the solution to the broth at a concentration of 5 g of ceftizoxime per ml. Next, we mixed 75 mg of aztreonam (Bristol-Myers Squibb) with 5 ml of distilled water, filtered the solution through an FP 030/2 filter (Schleicher & Schuell), and added it to the broth at a concentration of 75 g of aztreonam per ml. Finally, we filled sterile tubes with 8 ml of PHMB ϩ and stored them at 4°C in the dark. (The broth has a shelf life of at least 4 weeks.)We tested the PHMB ϩ with five different MRSA and five different methicillin-sensitive S. aureus (MSSA) strains isolated from patients. Methicillin resistance was confirmed by MecA PCR according to the method described by Murakami et al. (9). At first, all 10 strains were subcult...

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Comparison of a polymerase chain reaction assay and a conventional microbiologic method for detection of methicillin-resistant Staphylococcus aureus

Cited by 118 publications

References 18 publications

The evolutionary history of methicillin-resistant Staphylococcus aureus (MRSA)

The evolutionary history of methicillin-resistant Staphylococcus aureus (MRSA)

Performance of CHROM Agar and Oxacillin Resistant Screening Agar Base Media for Detection of Methicillin Resistant Staphylococcus aureus (MRSA) from Chronic Wound

Improved Detection of Methicillin-Resistant Staphylococcus aureus Using Phenyl Mannitol Broth Containing Aztreonam and Ceftizoxime

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