Misclassification of Susceptible Strains of
            <i>Staphylococcus aureus</i>
            as Methicillin-Resistant
            <i>S. aureus</i>
            by a Rapid Automated Susceptibility Testing System

Ribeiro, Julival; Vieira, Felipe do Nascimento; King, Tom; D’Arezzo, Julia B.; Boyce, John M.

doi:10.1128/jcm.37.5.1619-1620.1999

Cited by 34 publications

(18 citation statements)

References 17 publications

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“…Additionally, mecA transcriptional activity does not correlate with phenotypic methicillin resistance (31). Until new sets of recommendations are established, a combination of methods should be used routinely in detecting MRSA and OxRCoNS (8,12,35,48,49). A rapid PCR method that utilizes capillary air thermal cyclers to improve TAT has been published (7,19,20).…”

Section: Discussionmentioning

confidence: 99%

Rapid Extraction from and Direct Identification in Clinical Samples of Methicillin-Resistant Staphylococci Using the PCR

Jaffe

Lane

Albury

et al. 2000

Journal of Clinical Microbiology

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Methicillin-resistant staphylococci (MRS) are one of the most common causes of nosocomial infections and bacteremia. Standard bacterial identification and susceptibility testing frequently require as long as 72 h to report results, and there may be difficulty in rapidly and accurately identifying methicillin resistance. The use of the PCR is a rapid and simple process for the amplification of target DNA sequences, which can be used to identify and test bacteria for antimicrobial resistance. However, many sample preparation methods are unsuitable for PCR utilization in the clinical laboratory because they either are not cost-effective, take too long to perform, or do not provide a satisfactory DNA template for PCR. Our goal was to provide same-day results to facilitate rapid diagnosis and therapy. In this report, we describe a rapid method for extraction of bacterial DNA directly from blood culture bottles that gave quality DNA for PCR in as little as 20 min. We compared this extraction method to the standard QIAGEN method for turnaround time (TAT), cost, purity, and use of template in PCR. Specific identification of MRS was determined using intragenic primer sets for bacterial and Staphylococcus 16S rRNA and mecA gene sequences. The PCR primer sets were validated with 416 isolates of staphylococci, including methicillin-resistant Staphylococcus aureus (n ‫؍‬ 106), methicillin-sensitive S. aureus (n ‫؍‬ 134), and coagulase-negative Staphylococcus (n ‫؍‬ 176). The total supply cost of our extraction method and PCR was $2.15 per sample with a result TAT of less than 4 h. The methods described herein represent a rapid and accurate DNA extraction and PCR-based identification system, which makes the system an ideal candidate for use under austere field conditions and one that may have utility in the clinical laboratory.

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Section: Discussionmentioning

confidence: 99%

Rapid Extraction from and Direct Identification in Clinical Samples of Methicillin-Resistant Staphylococci Using the PCR

Jaffe

Lane

Albury

et al. 2000

Journal of Clinical Microbiology

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“…These subpopulations would be misclassified as susceptible rather than resistant S. aureus using phenotypic methods 26 . Hence, molecular detection of the mec A gene using polymerase chain reaction (PCR) is considered the gold standard for making a definitive diagnosis of methicillin resistance 26,28–31 …”

Section: Diagnosis Of Methicillin‐resistant Staphylococcal Infectionmentioning

confidence: 99%

A veterinary perspective on methicillin-resistant staphylococci

Cohn

Middleton

2010

Journal of Veterinary Emergency and Critical Care

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MRSA infection is an important cause of morbidity and mortality in humans. Animals may be contaminated, colonized, or infected with MRSA, with implications for the animal's health and as a potential reservoir for human infection. Staphylococci other than S. aureus may also acquire genes for methicillin resistance, and these species can also result in animal and occasionally human morbidity or mortality.

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“…Conventional cultural procedures for S. aureus identification include multiple subcultures and biotype or serotype identification steps, and require up to a week to obtain confirmed results (Francois et al 2006). Moreover, misidentification or misclassifications with automated susceptibility test systems or commercially available latex agglutination kits have been reported (Ribeiro et al 1999). Currently, various methods have been proposed for the detection and identification of S. aureus, including matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (Majcherczyk et al 2006), molecular-based techniques of principal component regression (PCR) assays (Francois et al 2003) and diagnostic marker analysis (Liu et al 2007).…”

Section: Introductionmentioning

confidence: 99%

RAPID IDENTIFICATION AND CLASSIFICATION OF STAPHYLOCOCCUS AUREUS BY ATTENUATED TOTAL REFLECTANCE FOURIER TRANSFORM INFRARED SPECTROSCOPY

Xie

et al. 2012

Journal of Food Safety

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Staphylococcus aureus is an important bacterium that can cause serious infections in human such as pneumonia and bacteremia. Rapid detection of this pathogen is crucial in food industries and clinical laboratories to control S. aureus food poisoning and human infections. In this study, Fourier transform infrared spectroscopy equipped with a germanium attenuated total reflection accessory was used as a novel approach to identify S. aureus. A total of 17 reference strains belonging to 4 different species and 84 clinical isolates of Staphylococcus spp. were analyzed. After the cultivation of the strains, spectral collection and data preprocessing, the S. aureus isolates were identified by a two‐step discrimination procedure. An internal validation and the related external validation were performed to demonstrate the discriminatory power and the quality of the discrimination models before the discrimination analysis. In the first step, 38 S. aureus isolates were correctly classified and the others were misidentified as Staphylococcus haemolyticus by hierarchical clustering analysis model using the first derivatives from the spectral range between 1,800 and 1,050/cm. In the second step, several classification/discrimination algorithms of soft‐independent modeling of class analogy, principal component regression and partial least squares regression (PLSR) were applied to build models for differentiating S. aureus and S. haemolyticus. The results showed that 57 (98.3%) strains and 4 (100%) strains of S. aureus and S. haemolyticus could be correctly identified by PLSR. PRACTICAL APPLICATIONS Fourier transform infrared (FTIR) spectroscopy is a potential method for rapid discrimination, classification and identification of intact microbial cells. In this study, FTIR spectroscopy equipped with a germanium attenuated total reflection accessory, using hierarchical clustering analysis–partial least squares regression discrimination analysis, is a powerful means for routine identification of Staphylococcus aureus.

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Misclassification of Susceptible Strains of Staphylococcus aureus as Methicillin-Resistant S. aureus by a Rapid Automated Susceptibility Testing System

Cited by 34 publications

References 17 publications

Rapid Extraction from and Direct Identification in Clinical Samples of Methicillin-Resistant Staphylococci Using the PCR

Rapid Extraction from and Direct Identification in Clinical Samples of Methicillin-Resistant Staphylococci Using the PCR

A veterinary perspective on methicillin-resistant staphylococci

RAPID IDENTIFICATION AND CLASSIFICATION OF STAPHYLOCOCCUS AUREUS BY ATTENUATED TOTAL REFLECTANCE FOURIER TRANSFORM INFRARED SPECTROSCOPY

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