scholarly journals Rapid Discrimination of Methicillin-Resistant Staphylococcus aureus by MALDI-TOF MS

Pathogens ◽  
2019 ◽  
Vol 8 (4) ◽  
pp. 214 ◽  
Author(s):  
Kim ◽  
Kim ◽  
Chung ◽  
Chung ◽  
Han ◽  
...  

Methicillin-resistant Staphylococcus aureus (MRSA) is a serious pathogen in clinical settings and early detection is critical. Here, we investigated the MRSA discrimination potential of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) using 320 clinical S. aureus isolates obtained in 2005–2014 and 181 isolates obtained in 2018. We conducted polymerase chain reactions (PCR) for staphylococcal cassette chromosome mec (SCCmec) typing and MALDI-TOF MS to find specific markers for methicillin resistance. We identified 21 peaks with significant differences between MRSA and methicillin-susceptible S. aureus (MSSA), as determined by mecA and SCCmec types. Each specific peak was sufficient to discriminate MRSA. We developed two methods for simple discrimination according to these peaks. First, a decision tree for MRSA based on six MRSA-specific peaks, three MSSA-specific peaks, and two SCCmec type IV peaks showed a sensitivity of 96.5%. Second, simple discrimination based on four MRSA-specific peaks and one MSSA peak had a maximum sensitivity of 88.3%. The decision tree applied to 181 S. aureus isolates from 2018 had a sensitivity of 87.6%. In conclusion, we used specific peaks to develop sensitive MRSA identification methods. This rapid and easy MALDI-TOF MS approach can improve patient management.

2021 ◽  
Vol 67 (2) ◽  
pp. 3372-3382
Author(s):  
Brigitta Horváth ◽  
Ferenc Peles ◽  
Judit Gasparikné Reichardt ◽  
Edit Pocklán ◽  
Rita Sipos ◽  
...  

The presence of methicillin-resistant Staphylococcus aureus (MRSA) strains in the food chain has been confirmed by several studies in the European Union, but there are only limited data available in Hungary. The objective of the present study was to investigate the antibiotic resistance of Staphylococcus strains isolated from foods, using classical microbiological, molecular biological methods and the MALDI-TOF-MS technique, as well as the multi-locus sequence typing (MLST) of antibiotic resistant strains. During the study, 47 coagulase-positive (CPS) and 30 coagulase-negative (CNS) Staphylococcus isolates were collected. In the course of the MALDI-TOF-MS investigations, all CPS isolates (n=47) were found to be S. aureus species, while 8 different species were identified in the case of the CNS strains. Methicillin resistance was confirmed in two S. aureus strains, one of which had a sequence type not yet known, while the other MRSA strain was type ST398, which is the most common type of MRSA strain isolated from farm animals in the EU/EEA. (The abbreviation “MRSA” is often used in common parlance, but occasionally in the literature to denote “multidrug-resistant Staphylococcus aureus”. In the authors’ manuscript - the methicillin-resistant pathogen is correctly designated as such. Ed.)


2011 ◽  
Vol 301 (1) ◽  
pp. 64-68 ◽  
Author(s):  
Manuel Wolters ◽  
Holger Rohde ◽  
Thomas Maier ◽  
Cristina Belmar-Campos ◽  
Gefion Franke ◽  
...  

Pathogens ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 430
Author(s):  
Wichai Santimaleeworagun ◽  
Praewdow Preechachuawong ◽  
Wandee Samret ◽  
Tossawan Jitwasinkul

Methicillin-resistant Staphylococcus aureus (MRSA) is mostly found in Thailand in the hospital as a nosocomial pathogen. This study aimed to report the genetic characterization of a clinical community-acquired MRSA (CA-MRSA) isolate collected from hospitalized patients in Thailand. Among 26 MRSA isolates, S. aureus no. S17 preliminarily displayed the presence of a staphylococcal cassette chromosome mec (SCCmec) type IV pattern. The bacterial genomic DNA was subjected to whole-genome sequencing. Panton–Valentine leukocidin (PVL) production, virulence toxins, and antibiotic resistance genes were identified, and multi-locus sequence typing (MLST) and spa typing were performed. The strain was matched by sequence to MLST type 2885 and spa type t13880. This strain carried type IV SCCmec with no PVL production. Five acquired antimicrobial resistance genes, namely blaZ, mecA, Inu(A), tet(K), and dfrG conferring resistance to β-lactams, lincosamides, tetracycline, and trimethoprim, were identified. The detected toxins were exfoliative toxin A, gamma-hemolysin, leukocidin D, and leukocidin E. Moreover, there were differences in seven regions in CR-MRSA no. S17 compared to CA-MRSA type 300. In summary, we have reported the ST2885-SCCmec IV CA-MRSA clinical strain in Thailand for the first time, highlighting the problem of methicillin resistance in community settings and the consideration in choosing appropriate antibiotic therapy.


2019 ◽  
Vol 12 (11) ◽  
pp. 1760-1768 ◽  
Author(s):  
Nimita Venugopal ◽  
Susweta Mitra ◽  
Rituparna Tewari ◽  
Feroze Ganaie ◽  
Rajeswari Shome ◽  
...  

Background and Aim: Methicillin-resistant staphylococci are among the emerging pathogens which have become a threat to both human and animal health. The present investigation intended to examine the occurrence and the molecular characteristics of methicillin-resistant Staphylococcus aureus (MRSA) and methicillin-resistant coagulase-negative staphylococci (MRCoNS) recovered from cattle, its handlers, and their environment. Materials and Methods: A total of 666 specimens were subjected to culture method and genus-specific polymerase chain reaction (PCR) for the identification of Staphylococcus. Methicillin resistance was substantiated by PCR identification of mecA and mecC resistance determinants. Species-specific identification of mecA positive isolates was conducted by multiplex PCR. The unidentified species were deciphered by 16S rRNA gene sequencing approach. The mecA positive isolates were further characterized by staphylococcal cassette chromosome mec (SCCmec) typing and multilocus sequence typing (MLST). Results: Duplex PCR identified 728 Staphylococcus isolates, of which 66 (9%) were positive for mecA gene. MRSA constituted 24% of the total mecA positive isolates. Among MRCoNS, Staphylococcus epidermidis (42%), and Staphylococcus haemolyticus (11%) were the most common species identified. Overall, 47% of the mecA positive isolates belonged to SCCmec type V. MLST analysis showed eight different sequence types (STs) among MRSA isolates of which five were novel STs. Among methicillin-resistant S. epidermidis, 19 different STs were found, of which nine novel STs were detected. Conclusion: The increase in the prevalence of mecA positive staphylococci, especially MRCoNS in cattle is a great concern in view of their transmission potential. Hence, continuous monitoring and molecular characterization of methicillin-resistant staphylococci should be elucidated in human and animal sectors so as to prevent the spread of these resistant pathogens.


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