Comparative and molecular analysis of MRSA isolates from infection sites and carrier colonization sites (original) (raw)
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Bosnian Journal of Basic Medical Sciences, 2015
Staphylococcus aureus is a major cause of hospital-acquired infections worldwide. Increased frequency of methicillin-resistant Staphylococcus aureus (MRSA) in hospitalized patients and possibility of vancomycin resistance requires rapid and reliable characterization of isolates and control of MRSA spread in hospitals. Typing of isolates helps to understand the route of a hospital pathogen spread. The aim of this study was to investigate and compare genotypic and phenotypic characteristics of MRSA samples on three different geography locations. In addition, our aim was to evaluate three different methods of MRSA typing: spa-typing, agr-typing and GenoType MRSA. We included 104 samples of MRSA, isolated in 3 different geographical locations in clinical hospitals in Zagreb, Mostar, and Heidelberg, during the period of six months. Genotyping and phenotyping were done by spa-typing, agr-typing and dipstick assay GenoType MRSA. We failed to type all our samples by spa-typing. The most common spa-type in clinical hospital Zagreb was t041, in Mostar t001, and in Heidelberg t003.We analyzed 102/104 of our samples by agr-typing method. We did not find any agr-type IV in our locations. We analyzed all our samples by the dipstick assay GenoType MRSA. All isolates in our study were MRSA strains. In Zagreb there were no positive strains to PVL gene. In Mostar we have found 5/25 positive strains to PVL gene, in Heidelberg there was 1/49. PVL positive isolates were associated with spa-type t008 and agr-type I, thus, genetically, they were community-associated MRSA (CA-MRSA). Dipstick assay GenoType MRSA has demonstrated sufficient specificity, sensibility, simple performance and low cost, so we could introduce it to work in smaller laboratories. Using this method may expedite MRSA screening, thus preventing its spread in hospitals.
Genotypic and phenotypic characteristics of Methicillin resistant Staphylococcus aureus MRSA strains
Staphylococcus aureus is a major cause of hospital-acquired infections worldwide. Increased frequency of methicillin-resistant Staphylococcus aureus (MRSA) in hospitalized patients and possibility of vancomycin resistance requires rapid and reliable characterization of isolates and control of MRSA spread in hospitals. Typing of isolates helps to understand the route of a hospital pathogen spread. The aim of this study was to investigate and compare genotypic and phenotypic characteristics of MRSA samples on three different geography locations. In addition, our aim was to evaluate three different methods of MRSA typing: spa-typing, agr-typing and GenoType MRSA. We included 104 samples of MRSA, isolated in 3 different geographical locations in clinical hospitals in Zagreb, Mostar, and Heidelberg, during the period of six months. Genotyping and phenotyping were done by spa-typing, agr-typing and dipstick assay GenoType MRSA. We failed to type all our samples by spa-typing. The most common spa-type in clinical hospital Zagreb was t041, in Mostar t001, and in Heidelberg t003.We analyzed 102/104 of our samples by agr-typing method. We did not find any agr-type IV in our locations. We analyzed all our samples by the dipstick assay GenoType MRSA. All isolates in our study were MRSA strains. In Zagreb there were no positive strains to PVL gene. In Mostar we have found 5/25 positive strains to PVL gene, in Heidelberg there was 1/49. PVL positive isolates were associated with spa-type t008 and agr-type I, thus, genetically, they were community-associated MRSA (CA-MRSA). Dipstick assay GenoType MRSA has demonstrated sufficient specificity, sensibility, simple performance and low cost, so we could introduce it to work in smaller laboratories. Using this method may expedite MRSA screening, thus preventing its spread in hospitals.
Molecular Epidemiology of Methicillin Resistant Staphylococcus Aureus (MRSA
Staphylococcus aureusis a gram-positive bacterium that can cause infections in almost anyone. The problem with the bacterium arises when S. aureus becomes resistant to methicillin. The bacterium's resistance to Methicillin and other beta-lactams is inherited from the mec.A gene located on the staphylococcal cassette chromosome mec (SCCmec), which serves a variety of functions in conferring its resistance to the drugs. It is important to understand the characteristics and components of infectious organisms, especially for epidemiological purposes. There are a variety of tests that can be conducted for the epidemiological and molecular study of MRSA which include plasma analysis, antibiogram typing, zerotyping, phage typing, bio typing, whole cell protein typing and zymotyping along with analysis restriction endonuclease. The two main DNA sequences analyzed when typing MRSA are MLST and SLST. MRSA can be classified as CA-MRSA (Community Acquired MRSA) and HA-MRSA (Hospital Acquired MRSA), based on the location where infection was contracted. Damage to skin and the mucous membrane gives a passage way for the bacteria to enter the individual. Diagnosis of MRSA can be made based on a samples obtained from an infected site on the patient and treatment is based on whether the infection is CA-MRSA or HA-MRSA. Worldwide, the infection rate of MRSA varies greatly, however, regardless of location, MRSA infection is a great concern. There are a variety of risk factors associated with infection of MRSA that have to be taken into consideration, especially when attempting to prevent the disease. This review looks into details of the molecular epidemiology of MRSA as well as the transmission, diagnosis and treatment options available.
Online journal of Health and Allied Sciences , 2024
Introduction: Methicillin-resistant Staphylococcus aureus (MRSA) has been known as an infectious pathogen worldwide since 1960. The epidemiological distribution of MRSA may have shifted due to outbreaks reported from several nations, making it more challenging to differentiate among CA-MRSA and HA-MRSA. It is currently important to develop a strain-based explanation for HA and CA-MRSA due to its distinct epidemiology, genetic profile, antibiogram, and quantifiable features. The study aimed to distinguish CA and HA-MRSA by Staphylococcal Cassette Chromosome mec (SCCmec) typing. Materials and Method: The study involved a total of 381 S. aureus isolates, which were processed in the department of Microbiology, JSS Hospital, Mysore. All isolates were confirmed as MRSA, initially by disk diffusion method using cefoxitin 30µg and oxacillin 1μg disk and later by using PCR technique for the detection of mecA-gene. All mecA-gene positive samples were amplified for SCCmec typing by multiplex PCR for detection of SCCmec type I, II, III, IVa, IVb, IVc, IVd, V and XI respectively. Results: PCR confirmed a total of 66% isolates as mecA-positive MRSA. Multiplex PCR method revealed only 53% isolates were SCCmec-typeable. The mainstream of the isolates belonged to SCCmec type IV (53.48%) and type V (44.18%), followed by type III (9.30%), type II (3%) and type I (1.16%) respectively. The study also demonstrated the presence of multiple SCCmec types in 10.46% of MRSA isolates. Staphylococcal cassette chromosome recombinase (ccr) typing determined only 43% of isolates were typeable. Conclusion: The study found that hospital-associated SCCmec type IV and type V were the most circulating strains in our healthcare setting. The research identified a few MRSA isolates with diverse SCCmec types. The presence of CA-MRSA infection in in-patients and HA-MRSA infection in This work is licensed under a Creative Commons Attribution-No Derivative Works 2.5 India License
Molecular identification and genotyping of MRSA isolates
Fems Immunology and Medical Microbiology, 2009
The aim of this study was to identify and characterize 97 methicillin-resistant Staphylococcus aureus (MRSA) isolates. Two conventional multiplex PCR assays, a real-time PCR assay and two PCR-based genotyping techniques including the spa- and hypervariable region (HVR)-typing methods were used to identify and characterize 97 MRSA strains isolated between April 2006 to September 2007 from the Steve Biko Academic Hospital. All MRSA isolates were positive for 16S rRNA gene, 99% were positive for the mecA gene and 4% positive for the Panton–Valentine leukocidin (PVL) gene. Staphylococcal cassette chromosome mec (SCCmec) typing showed 67% of isolates were SCCmec II [health-care-associated MRSA (HA-MRSA)], 14% were SCCmec III (HA-MRSA) and 4% were SCCmec IVd [community-associated MRSA (CA-MRSA)]. These CA-MRSA isolates showed a prevalence of 100% for the PVL gene. Using spa typing, three distinct clusters could be identified while HVR typing revealed six different clusters. CA-MRSA isolates were clustered together using spa and HVR typing. This study showed the prevalence of the CA-MRSA strains, PVL genes, the SCCmec types and the clonality of the MRSA strains. The high prevalence of the PVL gene in CA-MRSA isolates already residing in intensive care units was alarming and indicated the emergence of new MRSA lineages with a particular fitness for community and hospital transmission.
2020
Background: The global spread of methicillin-resistant Staphylococcus aureus (MRSA) infections necessitates the use of validated methods for the identification and typing of this bacterium. This study aimed to determine the distribution of main molecular types of MRSA strain circulating among hospitalized patients in teaching hospitals in Isfahan and Kashan. Methods: A total of 146 Staphylococcus aureus strains were isolated from patients in four teaching hospitals in Isfahan and Kashan during June 2017 to September 2018. The antibiotic resistance patterns of Staphylococcus aureus strains were performed by disc diffusion method. The MRSA strains were identified phenotypically and confirmed by PCR assay. The prevalence of microbial surface components recognizing adhesive matrix molecules (MSCRAMMs) genes among MRSA strains was evaluated by multiplex PCR. The genotypes of MRSA strains were determined by multi-locus sequence type and SCC mec typing. Results: Of 146 Staphylococcus aureus isolates 24 (16.4%) isolates identified as MRSA strains. According to antibiotic susceptibility testing the highest resistance rates were seen to erythromycin, cefoxitin and clindamycin. All of Staphylococcus aureus isolates were sensitive to vancomycin whereas 3 (2.1%) isolates were resistant to linezolid. Three different SCC mec types were obtained among MRSA strains including 16 (66.7%) SCC mec type V, 3 (12.5%) SCC mec type III and 5 (20.8%) SCC mec type II. Of 24 MRSA isolates 20 (83.3%) carried MSCRAMMs genes including eno (70.8%), fib (54.1%) , cna (25.0%), fnbB (16.6%) , ebps 5 (20.8%), and fnbA , bbp and clfA genes were not detected. MLST analysis revealed 11 sequence types among MRSA isolates as follows:
Background: Methicillin-resistant Staphylococcus aureus (MRSA) is considered one of the most important pathogenic bacteria and most prevalent pathogens causing dangerous infections in humans. Objectives: The purpose of this study was to analyze the hypervariable region (HVR) diversity of clinical MRSA isolates in Tabriz, northwestern Iran. Methods: In this retrospective and descriptive study, from Staphylococcus aureus strains isolated from clinical specimens of hospitalized patients from 2006 to 2013 at Tabriz health centers, 151 isolates were randomly selected. Methicillin-resistant isolates were identified by the agar disk diffusion method and mecA PCR assays. The genetic diversity of the isolates in the HVR were analyzed with the HVR typing method. Results: According to the antibiogram test results, from 151 samples, 52 isolates (34.4%) were resistant to cefoxitin. However, based on the polymerase chain reaction (PCR) assay, 54 isolates (35.8%) had the mecA gene and were identified as MRSA strains. According to PCR of the mecHVR, these MRSA strains were classified into seven different genotypes of HVR groups. Conclusions: High HVR diversity among the studied MRSA isolates could be a result of insufficient or inadequate infection-control protocols in Tabriz hospitals. Moreover, the high number of HVR genotypes showed that HVR typing can be used along with other typing methods in epidemiological studies of MRSA as a useful tool for monitoring, tracking contaminations, and controlling infections in hospital settings.
Infection, Genetics and Evolution, 2015
The burden of community-associated methicillin resistant Staphylococcus aureus (CA-MRSA) is on the rise in population and clinical settings on account of the adaptability and virulence traits of this pathogen. We characterized 45 non-duplicate CA-MRSA strains implicated mainly in skin and soft tissue infections (SSTIs) in a tertiary care hospital in Mysore, South India. All the isolates were genotyped by staphylococcal cassette chromosome mec (SCCmec) typing, staphylococcal protein A (spa) typing, accessory gene regulator (agr) typing, and multi-locus sequence typing (MLST). Four sequence types (STs) belonging to three major clonal complexes (CCs) were identified among the isolates: CC22 (ST2371 and ST22), CC1 (ST772) and CC8 (ST8). The majority (53.3%) of the isolates was of the genotype ST2371-t852-SCCmec IV [sequence type-spa type-SCCmec type], followed by ST22-t852-SCCmec IV (22.2%), ST772-t657-SCCmec V (13.3%) and ST8-t008-SCCmec IV (11.1%). ST237I, a single locus variant of ST22 (EMRSA-15 clone), has not been reported previously from any of the Asian countries. Our study also documents for the first time, the appearance of ST8-SCCmec IV (USA300) strains in India. Representative strains of the STs were further analyzed by pulsed field gel electrophoresis (PFGE). agr typing detected type I or II alleles in the majority of the isolates. All the isolates were positive for the leukotoxin gene, pvl (Panton-Valentine leukocidin) and the staphylococcal enterotoxin gene cluster, egc. Interestingly, multidrug resistance (resistance to P3 classes of non-beta-lactam antibiotics) was observed in 77.8% (n = 35) of the isolates. The highest (75.5%) resistance was recorded for ciprofloxacin, followed by erythromycin (53.3%), and quinupristindalfopristin (51.1%). Inducible clindamycin-resistance was identified in 37.7% of the isolates and it was attributed to the presence of erm(A), erm(C) and a combination of erm(A) and erm(C) genes. Isolates which showed a phenotypic pattern of M R /L S (macrolide-resistance/lincosamide-sensitivity) harbored the msr(A) gene. In conclusion, we report a high rate of multidrug resistance among Indian strains of CA-MRSA and the emergence of the lineages ST2371 and ST8 in India.
Prevalence and genotype distribution of MRSA in India
A B S T R A C T Methicillin-resistant Staphylococcus aureus (MRSA) is a serious human pathogen that can cause a wide variety of infections. Comparative genetic analyses have led to the discovery that despite the existence of a vast number of genotypes, outbreak strains of MRSA appear to be limited to certain genotypes, some of which are further restricted to certain geographical locations. Whereas extensive literature is available in several countries, the complexity of the clonal distribution both of healthcare-associated (HA) and community-associated (CA) MRSA in India is only now beginning to be understood. Studies have revealed that MRSA in India is distributed among all of the major staphylococcal cassette chromosome mec (SCCmec) types. The majority of HA-MRSA isolates belong to SCCmec type III and sequence type (ST) 239. By contrast, CA-MRSA mostly belong to ST22 (SCCmec IV), ST772 (SCCmec V) and ST672 (SCCmec V) genotypes. Similar to the global scenario, CA-MRSA is becoming more invasive and transmissible and is increasingly becoming difficult to be differentiated from HA-MRSA. In addition, it is disturbing that some of the HA-MRSA isolates have been reported to be vancomycin-resistant. On the other hand, almost no information is available on the genotypes of livestock-associated MRSA or their potential impact on human infections in India. Concerted efforts are needed to further understand the genetic epidemiology of MRSA in India.
Intisari Sains Medis
Background: Methicillin-Resistant Staphylococcus aureus (MRSA) is a big challenge for health services worldwide which causes infections both in healthcare and community. Healthcare-associated MRSA (HA-MRSA) strains are shown to be resistant to beta-lactam antibiotics and several non-beta lactam antibiotics. At the same time, the community-associated MRSA (CA-MRSA) tends to be resistant to beta-lactam antibiotics. MRSA carried staphylococcal cassette chromosome (SCCmec) types I, II, III, IV, and V. SCCmec types I, II, and III were predominantly found in HA-MRSA strain while SCCmec types IV and V predominantly found in CA-MRSA strains. Furthermore, the panton valentine leukocidine (pvl) gene is commonly found in CA-MRSA strains. Therefore, this study aimed to determine the prevalence of SCCmec types I, II, III, and pvl gene in MRSA isolated from clinical specimens in Sanglah General Hospital. Methods: This study was a cross-sectional descriptive study. MRSA was isolated from clinical...