Rapid Identification of
            <i>Staphylococcus aureus</i>
            and the
            <i>mecA</i>
            Gene from BacT/ALERT Blood Culture Bottles by Using the LightCycler System

Shrestha, Nabin K.; Tuohy, Marion J.; Hall, Gerri S.; Isada, Carlos M.; Procop, Gary W.

doi:10.1128/jcm.40.7.2659-2661.2002

Cited by 115 publications

(82 citation statements)

References 8 publications

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“…It has been reported earlier that mecA gene assay performed less optimally with the CoNS as compared to Staphylococcus aureus. 5 Moreover, PCR detected mecA gene in 4 isolates which were found to be methicillin sensitive by cefoxitin disk diffusion method. All these isolates, which were labeled as methicillin sensitive by the phenotypic method, should be regarded as potentially methicillin-resistant isolates bearing the mecA gene 6 and should not be classified as methicillin susceptible in spite of their susceptibility to beta lactam antibiotics.…”

Section: Discussionmentioning

confidence: 99%

“…2 mecA gene encodes for a particular PBP called PBP2A which has a low affinity for methicillin and most of the other b-lactam drugs and is consequently responsible for the intrinsic resistance to most of the b-lactams. 5 Since its initial detection in the 1960s, the incidence of infections caused by MR-CoNS is on the rise. 4 Kirby-Bauer disk diffusion test, agar or broth dilution and agar screen methods are the only standardized means of identifying methicillin resistance in the clinical microbiology laboratory.…”

Section: Discussionmentioning

confidence: 99%

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Prevalence and molecular characterization of methicillin resistance among Coagulase-negative Staphylococci at a tertiary care center

Bhatt

Tandel

Singh

et al. 2016

Medical Journal Armed Forces India

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m e d i c a l j o u r n a l a r m e d f o r c e s i n d i a 7 2 ( 2 0 1 6 ) s 5 4 -s 5Results: Out of 150 CoNS isolates, 51 were methicillin resistant by cefoxitin disk diffusion method. Out of these 51 isolates, mecA gene was detected only in 45 isolates. Moreover, mecA gene was also detected in 4 isolates, which were cefoxitin sensitive. Thus, the prevalence of methicillin resistance among CoNS was found to be 32.7% by PCR. Conclusion: The prevalence of methicillin resistance among Coagulase-negative Staphylococci (CoNS) was 32.7% by PCR detection of mecA gene. The sensitivity and specificity of cefoxitin disk diffusion method against mecA gene detection by PCR were found to be more than 90%. It can be concluded from this study that cefoxitin disk diffusion test can be used as a useful screening method to detect methicillin resistance among CoNS isolates. However, detection of mecA gene by PCR remains a more accurate method of detecting methicillin resistance among CoNS. #

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Prevalence and molecular characterization of methicillin resistance among Coagulase-negative Staphylococci at a tertiary care center

Bhatt

Tandel

Singh

et al. 2016

Medical Journal Armed Forces India

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“…Species-specific PCRs were used as the reference method in case of discrepant identification results (Table 2). Forty-six strains of 19 species (excluding meticillin-sensitive S. aureus, S. hyicus and S. xylosus) were identified using amplification and sequencing of the sodA gene (Martineau et al, 2000;Poyart et al, 2001;Shrestha et al, 2002;Stuhlmeier & Stuhlmeier, 2003;Morot-Bizot et al, 2004;Iwase et al, 2007;Hauschild & Stepanovic, 2008;Noguchi et al, 2010): Staphylococcus arlettae (n51), Staphylococcus capitis (n54), S. cohnii (n51), S. epidermidis (n53), S. haemolyticus (n56), S. hominis (n516), S. lugdunensis (n55), S. saprophyticus (n51), S. schleiferi (n52), Staphylococcus succinus (n55) and S. warneri (n52).…”

Section: Methodsmentioning

confidence: 99%

Evaluation of species-specific score cut-off values for various Staphylococcus species using a MALDI Biotyper-based identification

Richter

Hollstein

Woloszyn

et al. 2012

Journal of Medical Microbiology

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Although previous studies investigating the MALDI Biotyper database (matrix-assisted laser desorption/ionization time-of-flight mass spectrometry-based identification) have proven its high accuracy for bacterial identification, the studies differed in sample preparation, number of replicates, quantity of shots and target types used. In particular, the score cut-off values of special importance for reliable species identification varied. The aim of the present study was to identify species-specific differences in the mean score values for staphylococci. Cut-off values recommended by the manufacturer were adapted using the 20th percentile to rule out unknown score-modifying factors, even though the specificity was high and the lowest cut-off values would also yield an accurate result. Whilst correct species diagnosis was obtained in 97.32 % of samples (1382/1420), only 220 of all duplicates (15.49 %) revealed a score of ¢2.3, whilst 968 (68.17 %) had a score between 2.0 and 2.299, and 194 (13.66 %) had a score of ,2.0. Ten of 21 species had a calculated 20th percentile of ,2.0 and one species of ,1.7. In conclusion, the use of species-specific cut-off values improves the relative sensitivity of species identification in staphylococci.

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“…[20][21][22] Conventional PCR, and more recently real-time PCR, have been recognized as efficient tools for clinically applicable assays. 11,12,[23][24][25] The use of these tools is of particular interest in orthopedics, wherein the adhesion of bacteria within a biofilm on the surface of an implant may make the organism difficult to detect by conventional techniques. [15][16][17] Although some of these tools have been used to study orthopedic infections, much remains to be learned.…”

Section: Discussionmentioning

confidence: 99%

The comparison of pyrosequencing molecular Gram stain, culture, and conventional Gram stain for diagnosing orthopaedic infections

Kobayashi

Bauer

Tuohy

et al. 2006

Journal Orthopaedic Research

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ABSTRACT:We have developed a combined real-time PCR and pyrosequencing assay that successfully differentiated the vast majority of gram-positive and gram-negative bacteria when bacterial isolates were tested. The purpose of this study was to evaluate this assay on clinical specimens obtained from orthopedic surgeries, and to prospectively compare the results of ''molecular Gram stain'' with culture and conventional direct Gram stain. Forty-five surgical specimens were obtained from patients who underwent orthopedic surgery procedures. The DNA was extracted and a set of broad-range PCR primers that targeted a part of the 16S rDNA gene was used for pan-bacterial PCR. The amplicons were submitted for pyrosequencing and the resulting molecular Gram stain characteristics were recorded. Culture and direct Gram staining were performed using standard methods for all cases. Surgical specimens were reviewed histologically for all cases that had a discrepancy between culture and molecular results. There was an 86.7% (39/45) agreement between the traditional and molecular methods. In 12/14 (85.7%) culture-proven cases of bacterial infection, molecular Gram stain characteristics were in agreement with the culture results, while the conventional Gram stain result was in agreement only for five cases (35.7%). In the 31 culture negative cases, 27 cases were also PCR negative, whereas 4 were PCR positive. Three of these were characterized as gram negative and one as gram positive by this molecular method. Molecular determination of the Gram stain characteristics of bacteria that cause orthopedic infections may be achieved, in most instances, by this method. Further studies are necessary to understand the clinical importance of PCR-positive/culture-negative results. ß

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Rapid Identification of Staphylococcus aureus and the mecA Gene from BacT/ALERT Blood Culture Bottles by Using the LightCycler System

Cited by 115 publications

References 8 publications

Prevalence and molecular characterization of methicillin resistance among Coagulase-negative Staphylococci at a tertiary care center

Prevalence and molecular characterization of methicillin resistance among Coagulase-negative Staphylococci at a tertiary care center

Evaluation of species-specific score cut-off values for various Staphylococcus species using a MALDI Biotyper-based identification

The comparison of pyrosequencing molecular Gram stain, culture, and conventional Gram stain for diagnosing orthopaedic infections

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