The Characterization of mecA Gene and SCCmec Typing in Clinical Samples of MRSA

2019 ◽  
Vol 13 (2) ◽  
pp. 1
Author(s):  
Ali Jawad ◽  
Aziyah Abdul Aziz

MRSA is a major pathogen worldwide and its infections are associated with increased morbidity and mortality, in comparison with other S. aureus infections. The study was designed to characterize the mecA gene and staphylococcal cassette chromosome (SCCmec) in methicillin resistance S. aureus (MRSA). A total of 20 presumptive S. aureus strains were collected from one of the teaching hospitals in Selangor. Using standard biochemical tests, all the isolates were verified as S. aureus. When tested against cefoxitin, 80% of the isolates were confirmed as MRSA. All the MRSA isolates were further subjected to polymerase chain reaction (PCR) to detect the presence of mecA gene. Nine out of the 16 MRSA isolates (56%) were mecA positive, whereas the remaining four were mecA negative. The 16 MRSA with positive mecA gene were further subjected to SCCmec typing of type I, II, III, IV and V. The most frequent SCCmec types were type III (56%) followed by type II (33%), and type IV (11%). None of the isolates were of SCCmec type I or V. Our study indicates that SCCmec type III is predominant among the isolates which is in agreement with other studies conducted on clinical strains of MRSA.  

2003 ◽  
Vol 47 (11) ◽  
pp. 3574-3579 ◽  
Author(s):  
Hilmar Wisplinghoff ◽  
Adriana E. Rosato ◽  
Mark C. Enright ◽  
Michael Noto ◽  
William Craig ◽  
...  

ABSTRACT SCCmec is a mobile genetic element that carries the gene (mecA) mediating methicillin resistance in staphylococci. For Staphylococcus aureus, four SCCmec types have been described, one (type IV) of which has been associated with newly identified community-acquired methicillin-resistant S. aureus. However, the distribution of SCCmec types among S. epidermidis is not known. SCCmec typing of a collection of 44 methicillin-resistant Staphylococcus epidermidis (MRSE) isolates recovered between 1973 and 1983 from the blood of patients with prosthetic valve endocarditis (PVE) was performed by PCR amplification of key genetic elements (mecA, mecI, IS1272, and ccrAB). Of the 44 isolates, 1 (2%) harbored SCCmec type I, 15 (34%) harbored type II, 12 (28%) harbored type III, and 16 (36%) harbored type IV. The complete nucleotide sequence of SCCmec type IV was determined for 16 isolates and found to be identical in size (24 kb) and 98% homologous to DNA sequences published for S. aureus. Type IV SCCmec was also common (5 of 10 isolates) among a geographically dispersed collection of 10 recent (1998 to 2001) S. epidermidis bloodstream isolates. Multilocus sequence typing (MLST) (using the same seven genes presently employed for S. aureus MLST) of these MRSE isolates and of 10 additional recent geographically dispersed methicillin-susceptible isolates demonstrated that all 16 PVE isolates and 2 of 5 recent isolates harboring type IV SCCmec were in three related clonal groups. All three MSSE PVE isolates recovered from patients between 1976 and 1979 were in the same clonal groups as type IV SCCmec MRSE isolates. These data support the hypothesis of intra- and interspecies transfer of type IV SCCmec and suggest that there are clonal associations in S. epidermidis that correlate with SCCmec type.


2015 ◽  
Vol 8 (5) ◽  
Author(s):  
Hadi Sedigh Ebrahim-Saraie ◽  
Mohammad Motamedifar ◽  
Jamal Sarvari ◽  
Seyedeh Mahsan Hoseini Alfatemi

2021 ◽  
Vol 12 (3) ◽  
pp. 2207-2214
Author(s):  
Salama Mostafa Abdel-Hafez ◽  
Essam Hassan Mohamed ◽  
Youssef Saeed Alghamdi ◽  
Mohamed Mohamed Soliman ◽  
Saad Hommod Alotaibi ◽  
...  

The resistance of methicillin-resistant staphylococcus aureus (MRSA) to antimicrobials drugs is due to expression of the mecA gene. Current study was conducted on 33 MRSA clinical samples (Cefotaxime and Oxacillin positive). All MRSA isolates were examined using Antibiogram, Minimum Inhibitory concentration (MIC) and PCR to clarify the expression of SCCmec genes and to detect the differences on repeats of coagulase gene (Coa). Results showed that all isolates were 100% resistant against Amoxycillin-clavulanate, Ampicillin and Cefotaxime, 45.5% were resistant against Ciprofloxacin and erythromycin. mecA gene is expressed in all examined isolates (100%). The expression of SCCmec genes showed that 11.11% expressed type I, 45.45% contained type II, 45.45% contained type III, 63.63% were type IV and 5.55% were typeV. All examined isolates harbored and expressed coagulase gene repeats. Coagulase repeats were 27.27% with 5 repeats (81pb), and 72.72% with 4 repeats. In conclusion, the virulence of MRSA strains is increased and gave different antibiogram activities from different global regions and the repeats of Coa gene give no detectable differences among MRSA strains.


Author(s):  
Mahtab Sadat Madani Boroujeni ◽  
Mohammadreza Mahzounieh ◽  
Azizollah Ebrahimi Kahrizsangi ◽  
Soudabeh Rostami ◽  
Azam Mokhtari ◽  
...  

Background: Staphylococcus aureus (S. aureus) is a major cause of nosocomial infections in humans and animals. Because of the widespread resistance to antibiotics, microbiologists are trying to find other therapeutic interventions such as phage therapy for bacterial infections. Objectives: The present study aimed to isolate staphylophages with lytic effects on methicillin-resistant S. aureus (MRSA) clinical isolates as a potential alternative agent to antibiotic therapy. Methods: This experimental, descriptive study is performed in the Microbiology Laboratory of Shahrekord University (Iran) from September 2018 to March 2019. Two cocktails of staphylophages were isolated from Isfahan (Iran) urban sewage samples. The double-layer agar method was used to detect lytic phages. Morphology characteristic by transmission electron microscopy (TEM) images was used to identify staphylophages. One hundred and thirty three S. aureus were isolated from clinical samples of two teaching hospitals in Isfahan and Shiraz, Iran. Methicillin resistance and the presence of the mecA gene were determined by the disk diffusion method and polymerase chain reaction (PCR) assay, respectively. The phage susceptibility of mecA positive isolates was determined by plaque assay. Results: Two staphylophage cocktails were prepared, which had lytic effects on forty-four MRSA isolates. Cocktails 1 and 2 lysed 19 (14.2%) and 25 (18.7%) isolates, respectively. Of 133 S. aureus isolates, 88.7% carried the mecA gene. Conclusions: Different bacteriophages in two phage cocktails had relatively good lytic effects on S. aureus clinical isolates. Therefore, phage cocktails may be an appropriate alternative to antibiotics against S. aureus.


1999 ◽  
Vol 37 (5) ◽  
pp. 1426-1430 ◽  
Author(s):  
Atsuo Amano ◽  
Ichiro Nakagawa ◽  
Kosuke Kataoka ◽  
Ichijiro Morisaki ◽  
Shigeyuki Hamada

Fimbriae (FimA) of Porphyromonas gingivalis are filamentous components on the cell surface and are thought to play an important role in the colonization and invasion of periodontal tissues. We previously demonstrated that fimA can be classified into four variants (types I to IV) on the basis of the nucleotide sequences of the fimA gene. In the present study, we attempted to detect the four different fimA genes in saliva and plaque samples isolated from patients with periodontitis using the PCR method. Four sets of fimA type-specific primers were designed for the PCR assay. These primers selectively amplified 392-bp (type I), 257-bp (type II), 247-bp (type III), and 251-bp (type IV) DNA fragments of the fimA gene. Positive PCR results were observed with reference strains of P. gingivalis in a type-specific manner. All other laboratory strains of oral and nonoral bacteria gave negative results. The sensitivity of the PCR assay for fimAtype-specific detection was between 5 and 50 cells of P. gingivalis. Clinical samples were obtained from saliva and subgingival plaque from deep pockets (≥4 mm) of 93 patients with periodontitis. Bacterial genomic DNA was isolated from the samples, and the targeted fragments were amplified by PCR. The presence of P. gingivalis was demonstrated in 73 patients (78.5%), and a singlefimA gene was detected in most patients. The distribution of the four fimA types among the P. gingivalis-positive patients was as follows: type I, 5.4%; type II, 58.9%; type III, 6.8%; type IV, 12.3%; types I and II, 6.8%; types II and IV, 2.7%; and untypeable, 6.8%. P. gingivalis with type II fimA was detected more frequently in the deeper pockets, and a significant difference of the occurrence was observed between shallow (4 mm) and deep (≥8 mm) pockets. These results suggest that P. gingivalis strains that possess type II fimA are significantly more predominant in periodontitis patients, and we speculate that these organisms are involved in the destructive progression of periodontal diseases.


2020 ◽  
Vol 46 (02n03) ◽  
pp. 75-83
Author(s):  
Chiau-Jing Jung ◽  
Ya-Chih Huang ◽  
Jean-San Chia ◽  
Chih-Chieh Hsu ◽  
Chin-Cheng Chou

Staphylococcus pseudintermedius-induced soft tissue infection is a typical biofilm-related infectious disease. Recently, the occurrence of methicillin-resistant S. pseudintermedius (MRSP) infections in humans has increased. In this study, the virulence of different accessory gene regulator (agr) types of MRSP and MRSP[Formula: see text]induced skin infection were investigated through in vitro studies and the murine pyoderma model, respectively. Forty-seven canine clinical MRSP isolates were collected from northern Taiwan and classified into four agr types, namely, type I (52%), type II (2%), type III (35%), and type IV (10%). The majority of the MRSP isolates belonged to either strong (49%) or moderate (42.5%) biofilm producers. The skin damages induced by type III 77, type III 79, and type IV n10 isolates were larger than those induced by type I 58 and type II n3 isolates, but not by a significant degree. The effects of cell adhesion, cell invasion and cell cytotoxicity tests for agr type III 77, type III 79, and type IV n10 isolates were stronger than those caused by other types of MRSP isolates. However, the biofilm formation ability of these MRSP isolates did not show association with their virulence differences for causing murine skin abscesses. The agr type III 77, type III 79, and type IV n10 isolates of MRSP from the clinical samples revealed increased bacterial virulence for causing pyoderma, which is worthy of further genomic studies.


Author(s):  
G. D. Gagne ◽  
M. F. Miller ◽  
D. A. Peterson

Experimental infection of chimpanzees with non-A, non-B hepatitis (NANB) or with delta agent hepatitis results in the appearance of characteristic cytoplasmic alterations in the hepatocytes. These alterations include spongelike inclusions (Type I), attached convoluted membranes (Type II), tubular structures (Type III), and microtubular aggregates (Type IV) (Fig. 1). Type I, II and III structures are, by association, believed to be derived from endoplasmic reticulum and may be morphogenetically related. Type IV structures are generally observed free in the cytoplasm but sometimes in the vicinity of type III structures. It is not known whether these structures are somehow involved in the replication and/or assembly of the putative NANB virus or whether they are simply nonspecific responses to cellular injury. When treated with uranyl acetate, type I, II and III structures stain intensely as if they might contain nucleic acids. If these structures do correspond to intermediates in the replication of a virus, one might expect them to contain DNA or RNA and the present study was undertaken to explore this possibility.


2021 ◽  
Vol 8 (5) ◽  
pp. 85
Author(s):  
Yuttana Chanayat ◽  
Areerath Akatvipat ◽  
Jeff B. Bender ◽  
Veerasak Punyapornwithaya ◽  
Tongkorn Meeyam ◽  
...  

This study characterizes clinical methicillin-resistant staphylococcal (MRS) isolates obtained from superficial pyoderma infections in dogs. Our interest was to determine the staphylococcal cassette chromosome mec (SCCmec) type and the antimicrobial susceptibility among MRS isolates from clinical cases. Skin swabs were collected and cultured. Staphylococcus species were identified and characterized with biochemical tests and MALDI-TOF-MS and antimicrobial susceptibility testing by disk diffusion. mecA detection and staphylococcal cassette chromosome mec (SCCmec) typing were achieved by PCR. Of the 65 clinical samples, 56 (86.2%) staphylococcal infections were identified. Twelve (21%) of 56 isolates were MRS infections. All MRS isolates were multidrug resistant. The ccrC and class-C2 mec, which were SCCmec type V, were the most prevalent (66.7%) among the 12 MRS isolates. The predominant SCCmec type V was found in S. aureus, S. intermedius group, S. lentus, S. xylosus, and S. arlettae. Treatment failure is a concern with the emergence of highly resistant MRS in dogs associated with superficial pyoderma. The detection of type V SCCmec MRS has previously been reported among veterinarians and dog owners but not in Northern Thailand. These infections serve as a reminder to improve infection prevention and control measures including reducing environmental contamination and potential zoonotic exposures to MRS. In addition, educational awareness of these risks in small animal hospitals needs to be increased among veterinary hospital staff, clients, and patients.


Toxins ◽  
2020 ◽  
Vol 13 (1) ◽  
pp. 14
Author(s):  
Thamiris Santana Machado ◽  
Felipe Ramos Pinheiro ◽  
Lialyz Soares Pereira Andre ◽  
Renata Freire Alves Pereira ◽  
Reginaldo Fernandes Correa ◽  
...  

Hospitalizations related to Methicillin-resistant Staphylococcus aureus (MRSA) are frequent, increasing mortality and health costs. In this way, this study aimed to compare the genotypic and phenotypic characteristics of MRSA isolates that colonize and infect patients seen at two hospitals in the city of Niterói—Rio de Janeiro, Brazil. A total of 147 samples collected between March 2013 and December 2015 were phenotyped and genotyped to identify the protein A (SPA) gene, the mec staphylococcal chromosomal cassette (SCCmec), mecA, Panton-Valentine Leucocidin (PVL), icaC, icaR, ACME, and hla virulence genes. The strength of biofilm formation has also been exploited. The prevalence of SCCmec type IV (77.1%) was observed in the colonization group; however, in the invasive infection group, SCCmec type II was prevalent (62.9%). The Multilocus Sequence Typing (MLST), ST5/ST30, and ST5/ST239 analyses were the most frequent clones in colonization, and invasive infection isolates, respectively. Among the isolates selected to assess the ability to form a biofilm, 51.06% were classified as strong biofilm builders. Surprisingly, we observed that isolates other than the Brazilian Epidemic Clone (BEC) have appeared in Brazilian hospitals. The virulence profile has changed among these isolates since the ACME type I and II genes were also identified in this collection.


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.


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