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Study of Staphylococcus aureus genotyping using Coagulase gene method
Abstract
Fifty isolates of Staphylococcus aureus bacteria were obtained out of 200 samples collected from clinical sources. The highest resistance to S. aureus bacteria was 100% Benzylpericillin, Oxacillin and Moxiflaxocin antibiotics, 86% Levoflaxacin resistance, 76% Erythromycin resistance, 66% Tetracycline resistance, 64% Tobramycin resistance, 62% Gentamicin resistance, 60% Clindamycin resistance and 58% Trimethoprim/Sulfamethoxazole resistance. It was 38% sensitive to Gentamicin and Clindamycin, 36% sensitive to Tobramycin, 22% sensitive to Erythromycin and 14% sensitive to Levoflaxacin while the results showed the presence of five different sized types of DNA packets when detecting Coa gene of S.aureus bacteria using PCR techniques.
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The results of the present study showed that twenty-five samples were collected for the age group 35–40 years and four samples for the age group 65–70 years for both genders. The results showed that 48 (48%) of the samples were obtained from the hands, 16 (16%) from the legs, 12 (12%) from the abdominal area, and 10 (10%) from the chest area. The four (4%) samples were obtained from burns in the back and thighs area. The samples taken according to the cause of burns were 40 (40%) due to hot water, hot liquids, or hot steam, followed by 18 (18%) due to the use of hot tools, 15 (15%) due to fires, 12 (12%) due to electric currents, 10 (10%) due to chemicals such as strong acids, alkaline lye, paint thinner, or gasoline, and 5 (5%) due to sun ray burns. Sixty pathogenic bacteria were obtained from the burn samples. The number of bacteria isolated from burn wounds was 34 isolates from men and 26 isolates from women. The predominant were 15 (25%) Staphylococcus aureus, 12 (20%) Acinetobacter baumannii, 10 (16.7%) Pseudomonas aeruginosa, 8 (13.3%) Klebsiella pneumoniae, 7 (11.7%) Escherichia coli, 6 (10%) Proteus mirabilis, and 2 (3.3%) Burkholderia cepacia. The antibiotic sensitivity test using the Vitek2 Compact System showed that the resistance rate was recorded in Staphylococcus aureus against Amikacin by 13 isolates, with a rate of 86.6%, and in Acinetobacter baumannii, towards Ceftazidime and Piperacillin antibiotics by 12 isolates at a rate of 100%, and Pseudomonas aeruginosa towards Colistin and Tobramycin at a rate of 6 isolates at a rate of 60%, and Klebsiella pneumoniae towards Colistin and Tobramycin at a rate of 8 isolates at a rate of 100% and Escherichia coli against Amikacin, Colistin, and Imipenem with 7 isolates and 100%, and Proteus mirabilis against Colistin and Tobramycin with 6 isolates and 100%, and Burkholderia cepacia against 8 antibiotics with a rate of 100%. We conclude from the present study that the most susceptible age group to burns is the active age group and that the pathogenic bacteria from burn wounds are mostly resistant to antibiotics.
Staphylococcus aurous is common pathogens of the eye include inflammation of Conjunctivitis. Slime forming and antibiotics resistance currently consider as major virulence factors. The aim of the present study was Isolation and identification of biofilm forming S. aurous and screening the prevalence icaA gene. A total of 50 conjunctiva swap samples have been collected from patients clinically diagnosed with bacterial infection conjunctivitis in two hospitals at Baghdad city, include Al-Furat General Hospital and Ibn Al-Haitham Teaching Hospital during the period of study from the beginning of November 2019 to the end of April 2020. among 50 swap specimen, 22 (44%) specimen were given positive results for bacterial infections, while 28 (56%) specimen were given negative results, which mean the infection caused by other agents. However among 22 positive bacterial isolation swap (12) samples were given positive results for bacterial infections, as (Pseudomonas, Streptococcus, Klebsiella spp, Escherichia coli, Proteus spp), while (10) specimen were Staphylococcus species. The predominant isolates were Staphylococcus aurous about 6(60%), depending on the morphologic characteristics of this bacterium on the culture media and biochemical tests included manual conventional biochemical tests and automated biochemical tests such as Vitek 2 test. Molecular detection conducted using Polymerase Chain Reaction technique of icaA gene with 603 bp. In addition by recA was used to confirm that these isolates were belonging to S. aurous.
Three hundred and sixty different samples were collected from different sources, including wound, burn, nasal, and oral swabs from several hospitals in Baghdad. A number of 150 (53%) Staphylococcus aureus samples were isolated and identified among a total of 283 samples. Then, the spread of the Toxic Shock Syndrome Toxin-1 gene (tsst-1) was investigated in β-lactamase resistant S. aureus. According to the source of samples, the distribution of S. aureus isolates was found to be significantly higher (p < 0.01) in wound samples as compared to other sources. According to the age, a highly significant distribution (p < 0.01) was recorded in the age group of 15-30 years, whereas gender comparison showed no statistically significant differences. All the isolates were subjected to susceptibility test against eight β-Lactam antibiotics by using the disc diffusion method. The antimicrobial susceptibility test showed that S. aureus had maximum resistance percentage to Carbenicillin (97.3%), while the lowest resistance rate was against Meropenem, with a sensitivity rate of up to 82%. In addition, 144 (96%) out of the 150 S. aureus isolates have multiple drug resistance (MDR). All the isolates were subjected to polymerase chain reaction to amplify tsst-1 toxin gene. A number of 70 isolates (46.7%) were found to be positive for tsst-1 gene. The results showed no significant correlation between tsst-1 gene with the individual antibiotic resistance and the multi-drug resistance patterns of the isolates (p = 0.226).
A total 248 clinical samples of cerebrospinal fluid (CSF) were collected from meningitis patients, for the period of July to October 2018 from the Child Protection Teaching Hospital in the Medical City Compound in Baghdad. All isolates were identification depending on macroscopic, microscopic, biochemical tests and definite with Vitek-2 compact system. The results showed a growth in 42 samples. Sixteen isolates of Staphylococcus spp. were obtained from samples. The number (percentage) of isolates according to species as follow: 7/16) 44%) of S. epidermidis, 3/16 (19%) of S. hominis, 3/16) 19%) of S. haemolyticus, 2/16 (12%) of S. aureus, 1/16 (6%) of S.warneri. These isolates were tested for ability to biofilm formation by using Luria Broth and Tryptone Soya Broth (TSB) supplemented with 0.25% and 2% glucose. The ability of the isolates to form biofilm with Luria Broth, for S.epidermidis, S. aureus, S. haemolyticus, S.hominis and S. warneri were 7/7 (100%), 1/2 (50%), 2/3 (66.66%), 1/3 (33.33%),1/1 (100%) respectively not forming biofilm.As for using a TSB supplemented with 0.25% glucose, S.epidermidis, S. aureus, S. haemolyticus and S.hominis were 6/7 (85.71%), 2/2 (100%), 2/3 (66.66%) and 3/3 (100%) respectively were weakly biofilm formed and when using TSB supplemented with 2% glucose, showed that all Staphylococcus spp. (100%) were positive for the test..The study showed the effect of minimum inhibition concentration(8 μg/ mL) and minimum bactericidal concentrationand (16μg/mL) of Erythromycin on biofilm of Staphylococcus spp. Staphylococcus spp. isolated from CSF showed weak ability to biofilm formation and depending on the host content of sugars. This study showed also weak biofilm forming of Staphylococcus spp. was resistence to erythromosin.
Staphylococcus epidermidis, Staphylococcus haemolyticus and Enterococcus gallinarum bacteria were obtained from the vaginal infection, during the period from July to October from Azadi from Azadi Teaching Hospital in Kirkuk, Iraq. The effect of menthol and thymol on bacterial growth was studied using concentration (300-3100 µg). The compounds showed an antibacterial activityat all concentrations, as well as the effect of the compounds on the biofilm formation. Menthol showed a high activity in inhibiting the formation of the biofilm of S. haemolyticus and E. galinarum from thymol and studied combination of menthol-thymol with a concentration ofmenthol (1500 and 700 µg/ml), thymol (700, 300 µg/ml) for inhibiting the growth of isolated bacteria compared to the two components alone.
The resistance is determining a growing hug issue in health fields worldwide, where the bacterial cells possessed the ability to resist the old antibiotics as well as the newly discovered antibiotics through several capabilities and mechanisms, including the natural, acquired and cross ones, this paper will highlight most of the antibiotics and the mechanics of resistance.
BackgroundS.aureus is a predominant pathogen that causes infection in critically ill patients, but little information exists regarding the characterization of S. aureus from different sources in burn patients in southeastern China. Methods
We enrolled 125 patients with S. aureus infection in burns center between Jan 2014 and Dec 2015. S. aureus isolates were characterized by antimicrobial susceptibility test, toxin gene detection, and molecular typing with multilocus sequence type, staphylococcal protein A (spa) type, and staphylococcal cassette chromosome mec (SCCmec) type. ResultsSixty-eight MRSA were isolated from SSTI and 31 from non-SSTI patients, respectively. Overall, the drug-resistant ability of S. aureus isolated from SSTI was higher than that from non-SSTI groups. SCCmecIII-CC239-t030 was the most common clone (38 from SSTIs, and 8 from non-SSTIs). Seg was the most common enterotoxin gene (21 from SSTIs and 33 from non-SSTIs). Isolates from SSTIs was more likely to carry seb (P = 0.04), while those from non-SSTIs tended to carry sea and seg (P = 0.002 and 0.01, respectively). Although isolates carried four hemolysin genes, there was no significant difference between them (P > 0.05). ConclusionSCCmecIII-CC239-t030 was the most common clone in Jiangxi burns center, China. The molecular characterization of S. aureus was quite different between SSTI and non-SSTI groups.
Background: Staphylococcus aureus has become an emerging public health concern. Markers that differentiate tissue-specific lineages are needed to trace the sources of strains. Objectives: The aims of this study were to determine the genotypic characteristics of S. aureus isolates that are associated with skin and urinary tract infections using polymorphisms in the coagulase gene. Materials and Methods: Coagulase gene variants among 26 S. aureus isolates from human infected skin (n = 10) and urine (n = 16) samples were investigated by amplification of the repeat units encoding the hypervariable region of the coagulase gene. The amplicons ranged from 490-790 bp and were subjected to restriction fragment length polymorphism (RFLp) analysis with Haelll. Results: In total, 6 distinct RFLp banding patterns were observed, designated C1-C6. The C1 pattern predominated in skin and urine isolates. Notably, the C3, C5, and C6 patterns were present in isolates from urine, whereas the C2 and C4 genotypes were preferentially detected in skin sample isolates. Conclusions: These data demonstrate the widespread prevalence of certain genotypes and tissue-specific tendency of other genotypes, suggesting the existence of lineage- and tissue-specific genes that mediate the development of tissue-specific pathogenicities of S. aureus isolates. Copyright
To determine the prevalence and sensitivity trends of urinary bacterial isolates.
A cross-sectional study.
Gondar College of Medical Sciences (GCMS) Teaching and Referral Hospital, north west Ethiopia.
Four hundred and twenty urine specimens from 70 in-patient and 350 out-patient cases were studied by quantitative culture method and anti-microbial sensitivity test was done by disc diffusion technique.
One hundred and seventy two pathogenic organisms were isolated from 166 patients; the isolation rate was 39.5 %. Among the isolates E. coli, S. aureus, Klebsiella species, coagulase negative Staphylococcus species and Citrobacter species were common accounting for 46.0%, 18.0%, 10.0%, 8.0% and 6.0%, respectively. Of the total isolates 71.5% were Gram negatives. Sensitivity tested against ten antibiotics showed that resistance was common, and the effectiveness of tetracycline, ampicillin, co-trimoxazole, chloramphenicol and penicillin was under 50.0%. The resistance rate was 71.5%, 62.2%, and 62.2%, 54.7% and 40.8%, respectively. Polymixin B, cefoxitin, gentamycin and erythromycin controlled over 76.0% of the common infective agents. Ciprofloxacin did control 98.3% of the organisms.
Resistance was found to be very high to the commonly used antibiotics. The sensitivity rate for the recently introduced ciprofloxacin was above 98%. Therefore, this antibiotic may be used for empirical therapy of urinary tract infection (UTI) when culture and sensitivity testing is impossible. Strict control on the use of antibiotics and appropriate measures against over the counter availability and self-medication is recommended.
Acinetobacter baumannii has emerged as a major cause of both community-associated and nosocomial pneumonia, but little is known about the cellular
and molecular mechanisms of host defense against respiratory infection with this bacterial pathogen. In this study, we examined
the role of neutrophils in host resistance to pulmonary A. baumannii infection in a mouse model of intranasal (i.n.) infection. We found that neutrophils were rapidly recruited to the lungs
following i.n. inoculation of the pathogen and declined to baseline level upon clearance of the infection. Depletion of neutrophils
using monoclonal antibody RB6-8C5 prior to infection resulted in an acute lethal infection that was associated with enhanced
bacterial burdens in the lung (P < 0.05) and extrapulmonary dissemination to the spleen. The increased susceptibility to A. baumannii in neutropenic mice was associated with a delay in the mRNA expression and production of early proinflammatory cytokines
such as tumor necrosis factor alpha, interleukin-6, keratinocyte chemoattractant protein, monocyte chemoattractant protein
1, and macrophage inflammatory protein 2 (MIP-2) in the lungs and development of severe bronchopneumonia and lymphoid tissue
destruction in the spleen. Moreover, i.n. administration of the neutrophil-inducing chemokine MIP-2 to normal mice induced
a pulmonary influx of neutrophils and significantly enhanced the clearance of A. baumannii from the lungs (P < 0.01). These results imply that neutrophils play a critical role in host resistance to respiratory A. baumannii infection.
Bacterial vaginosis (BV) is a common infection that occurs when the number of lactobacillus spp. bacteria (vaginal flora) decreases in the vaginal canal. The study aimed to determine the prevalence of Staphylococcus aureus within vaginosis in order to emphasize the importance of early detection and treatment. Totally, 90 vaginal swabs were collected using speculum and swabbing. The vaginal swabs were subjected to standard microbiological testing, which included microscopy, cultures (Blood agar and Mannitol salt agar), and antibiotic sensitivity testing. The results showed that out of 90 samples, only 40 S.aureus isolates were collected. S. aureus isolates showed maximum sensitivity to gentamicin and high resistance to Cefoxitin, Erythromycin, Azithromycin, and moderate resistance to Clindamycin, Rifampicin, Ciprofloxacin, and Doxycycline. Regarding molecular detection of mecA, the results revealed that out of 40 S. aureus isolates only 22 isolates have this gene. This study concludes that S. aureus (MRSA) was the most prevalent within vaginosis with high resistance to most antibiotics.
Tonsillitis is an infection of the tonsils caused by one of several types of bacteria or viruses. Staphylococcus aureus are one of most common bacteria isolated from recurrent tonsillitis, its identification by classical methods takes 3-5 days as well as the accuracy does not reach an absolute degree. So current study aimed to diagnose S. aureus by classical and molecular diagnosis to increase the accuracy of the diagnosis. Two hundred and fifteen tonsil swabs were collected from Iraqi patients susceptible suffered from recurrent tonsillitis who attended clinics in major hospitals in Najafe/ Iraq. Classical diagnoses of S. aureus were done by bacterial cultures, biochemical test and Vitek system. While molecular diagnosis achieved via PCR technique The results of classical methods showed that 50 isolates recovered from (195) suspected patients were S. aureus, while molecular diagnoses showed that 45 isolates out of 50 diagnostic S. aureus via classical methods were S. aureus with significant difference (P≤ 0.01). In conclusion, Molecular method is more sensitive in diagnoses of S. aureus than conventional testes in case of recurrent tonsillitis.
Abstract: Coagulase is considered as the main determinant to distinguish Staphylococcus aureus. The 3′ end coding region of the coagulase gene (coa) has a series of 81bp tandem repeats varying in number and sites of restriction enzyme among diverse isolates. The polymorphism of the coagulase gene among 45 MRSA isolated from burn wounds, otitis media and pneumonia in Basra city, Iraq, were investigated using PCR-RFLP analysis for 3′ end region with AluI enzyme revealing unique restriction patterns (18 patterns) not described in any previous studies. Particularly, pattern 1 was predominant. Different RFLP patterns of coa were shared among isolates from the different sampling sites.
The present study used the polymorphism of Coagulase gene to identify MRSA subtypes, estimate the efficiency of these methods in distinguishing the variable strains and compare these subtypes with the source of isolates.
This study was designed to determine the percentage and the main causative agent causing bacteremia among children aged up to 12 years and complaining from different types of infections (Respiratory, intestinal, and urinary tract infection) in Baghdad. Results showed that the percentage of infection was 46.19 % the main causative agents were Enterobacteriaceae including (E.coli , Pseudomonas , Salmonella.typhi .Serratia , Enterobacter , Klebsiella )and other than Enterobacteriaceae which includes(Staph.aureus , Staph.epidermidis , Streptococcus.Pneumonia and ?-hemolytic streptococci ) .Regarding the age factor ,results showed that the highest infection rate was among the age group (1 day-12 month ) and (12 -36month ) (64.89%)and (15.95%)respectively while the lowest was in (61 month – 12 years )and (37 -60 month) (12.76%) and( 6.38) respectively . All bacterial strains isolated from patient were submitted to sensitivity test, results showed various reactions towards different types of antibiotics used in this study.
Quinolone antimicrobials are synthetic and widely used in clinical medicine. Resistance emerged with clinical use and became common in some bacterial pathogens. Mechanisms of resistance include two categories of mutation and acquisition of resistance-conferring genes. Resistance mutations in one or both of the two drug target enzymes, DNA gyrase and DNA topoisomerase IV, are commonly in a localized domain of the GyrA and ParE subunits of the respective enzymes and reduce drug binding to the enzyme-DNA complex. Other resistance mutations occur in regulatory genes that control the expression of native efflux pumps localized in the bacterial membrane(s). These pumps have broad substrate profiles that include quinolones as well as other antimicrobials, disinfectants, and dyes. Mutations of both types can accumulate with selection pressure and produce highly resistant strains. Resistance genes acquired on plasmids can confer low-level resistance that promotes the selection of mutational high-level resistance. Plasmid-encoded resistance is due to Qnr proteins that protect the target enzymes from quinolone action, one mutant aminoglycoside-modifying enzyme that also modifies certain quinolones, and mobile efflux pumps. Plasmids with these mechanisms often encode additional antimicrobial resistances and can transfer multidrug resistance that includes quinolones. Thus, the bacterial quinolone resistance armamentarium is large.
© 2015 New York Academy of Sciences.
From 1965 to 1973, 75 patients were admitted to the Children's Hospital of Buffalo, with toxic epidermal necrolysis (TEN). A review of their records show an equal ratio of males to females, with 73 whites and only 2 blacks. No patient was septic with staphylococci, and there was only one death. A comparison of treatments indicates that the disease is probably self-limited and not influenced by antibiotics or corticosteroids.
To ascertain the clinical importance of a strain of slide coagulase positive but tube coagulase negative Staphylococcus species isolated from the blood culture of a 43 year old patient with refractory anaemia with excessive blasts in transformation who had neutropenic fever.
The isolate was investigated phenotypically by standard biochemical methods using conventional biochemical tests and two commercially available systems, the Vitek (GPI) and API (Staph) systems. Genotypically, the 16S ribosomal RNA (rRNA) gene of the bacteria was amplified by the polymerase chain reaction (PCR) and sequenced. The sequence of the PCR product was compared with known 16S rRNA gene sequences in the GenBank by multiple sequence alignment.
Conventional biochemical tests did not reveal a pattern resembling a known Staphylococcus species. The Vitek system (GPI) showed that it was 94% S. simulans and 3% S. haemolyticus, whereas the API system (Staph) showed that it was 86.8% S. aureus and 5.1% S. warneri. 16S rRNA gene sequencing showed that there was a 0 base difference between the isolate and S. aureus, 28 base difference between the isolate and S. lugdunensis, 39 base difference between the isolate and S. schleiferi, 21 base difference between the isolate and S. haemolyticus, 41 base difference between the isolate and S. simulans, and 23 base difference between the isolate and S. warneri, indicating that the isolate was a strain of S. aureus. Vancomycin was subsequently prescribed and blood cultures taken four days after the start of treatment were negative.
16S rRNA gene sequencing was useful in ascertaining the clinical importance of the strain of slide coagulase positive but tube coagulase negative Staphylococcus species isolated from blood culture and allowing appropriate management.
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