scholarly journals Immunofluorescence Targeting PBP2a Protein: A New Potential Methicillin Resistance Screening Test

2021 ◽  
Vol 8 ◽  
Author(s):  
Serenella Silvestri ◽  
Elisa Rampacci ◽  
Valentina Stefanetti ◽  
Michele Trotta ◽  
Caterina Fani ◽  
...  
Keyword(s):  
Diagnostic Accuracy ◽  
Screening Test ◽  
Methicillin Resistance ◽  
Protein A ◽  
Resistant Bacteria ◽  
Bacterial Cells ◽  
Resistance Screening ◽  
Culture Methods ◽  
Easy Method ◽  
Apparent Prevalence

The indiscriminate use of first-line drugs contributed to the spread of resistant bacteria, a major concern for both human and veterinary medicine. Methicillin resistance is acquired through the mecA gene, which encodes for the PBP2a protein and lends the resistance to β-lactams. Verifying the correspondence between gene harboring and protein expression and accelerating methicillin resistance diagnosis is critical to improve the management of antimicrobial administration and to reduce the spread of drug resistances. We tested the applicability of immunofluorescence targeting PBP2a protein to identify a new potential methicillin resistance screening test, ancillary to conventional culture methods. We collected 26 clinical Staphylococcus pseudintermedius (SP) isolates: 25 from canine pyoderma and 1 from dermatitis in a dog owner. SP is one of the most important etiological agents in canine pyoderma and can harbor the mecA gene. We performed PCR for mecA gene detection, broth microdilution (BMD) for phenotypic methicillin resistance, and immunofluorescence targeting PBP2a protein. Compared to the PCR as the gold standard, immunofluorescence showed an apparent prevalence of 34.6% vs. a true prevalence of 53.8%, with 100% specificity, 64.3% sensitivity, and 80.8% diagnostic accuracy. PBP2a expression showed isolate-dependent variability: in some isolates, most of the bacterial cells showed an intense and clearly membranous pattern, while in others only a few of them could be detected. Performing the assay in duplicate improved the diagnostic accuracy. Since the mecA gene is shared among the members of the Staphylococcus genus, the test can be applied to identify methicillin resistance independently from the staphylococcal species, both in human and animal samples. Being a rapid and easy method and providing the unique possibility to study the expression of PBP2a by directly visualizing the morphology, it could represent a new interesting tool for both research and diagnostics. To accelerate methicillin resistance diagnosis, it would be worth further testing of its performance on cytological samples.

The comparative study among the MRSAcin, Nisin A and vancomycin, on biofilm formation by Methicillin resistance Staphylococcus aureus isolated from food sources.

2019 ◽  
Vol 9 (o3) ◽  
Author(s):  
¹Hind H. Muunim ◽  
Muna T Al-Mossawei ◽  
Mais Emad Ahmed
Keyword(s):  
Biofilm Formation ◽  
Methicillin Resistance ◽  
Microbial Control ◽  
Antibiotic Sensitivity ◽  
Raw Milk ◽  
Sensitivity Test ◽  
Food Sources ◽  
Resistant Bacteria ◽  
Biochemical Tests ◽  
Food Preservatives

Biofilms formation by pathogens microbial Control considered important in medical research because it is the hazarded virulence factor leading to becoming difficult to treat because of its high resistance to antimicrobials. Glycopeptide antibiotic a (Vancomycin) and the commercial bacteriocin (Nisin A) were used to comparative with purification bacteriocin (MRSAcin) against MRSA biofilm. One hundred food samples were collected from Baghdad markets from July 2016 to September 2016, including (cheese, yogurt, raw milk, fried meat, grilled meat, and beef burger). All samples were cultures; S. aureus was confirmation by macroscopic culture and microscopic examination, in addition to biochemical tests. Methicillin resistance S. asureus (MRSA) were identification by antibiotic sensitivity test (AST), Vitek 2 system. The result shown the 60(60%) isolate were identified as S. aureus and 45(75%) gave positive result as MRSA isolate, M13 isolate was chosen as MRSA isolates highest biofilm formation for treatment with MRSAcin, Nisin A(bacteriocin) and Vancomycin (antibiotic) to compared the more antimicrobial have bacteriocidal effect. The sensitivity test uses to determine the effect of MRSAcin, Nisin A, and Vancomycin MIC on MRSA planktonic cell by (WDA). The new study shows the impacts of new kind Pure Bacteriocins (MRSAcin) from methicillin-resistant S. aureus (MRSA) highly effects then (Vancomycin and Nisin A) at different concentration. In a current study aimed to suggest new Bacteriocin is potent highly for the treatment of resistant bacteria biofilm infections in food preservatives

scholarly journals Liposomes as Antibiotic Delivery Systems: A Promising Nanotechnological Strategy against Antimicrobial Resistance

Molecules ◽  
2021 ◽  
Vol 26 (7) ◽  
pp. 2047
Author(s):  
Magda Ferreira ◽  
Maria Ogren ◽  
Joana N. R. Dias ◽  
Marta Silva ◽  
Solange Gil ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Bacterial Infections ◽  
Private Investment ◽  
Therapeutic Index ◽  
Multidrug Resistant ◽  
Current Treatment ◽  
Delivery Systems ◽  
Resistant Bacteria ◽  
Bacterial Cells ◽  
Antimicrobial Drugs

Antimicrobial drugs are key tools to prevent and treat bacterial infections. Despite the early success of antibiotics, the current treatment of bacterial infections faces serious challenges due to the emergence and spread of resistant bacteria. Moreover, the decline of research and private investment in new antibiotics further aggravates this antibiotic crisis era. Overcoming the complexity of antimicrobial resistance must go beyond the search of new classes of antibiotics and include the development of alternative solutions. The evolution of nanomedicine has allowed the design of new drug delivery systems with improved therapeutic index for the incorporated compounds. One of the most promising strategies is their association to lipid-based delivery (nano)systems. A drug’s encapsulation in liposomes has been demonstrated to increase its accumulation at the infection site, minimizing drug toxicity and protecting the antibiotic from peripheral degradation. In addition, liposomes may be designed to fuse with bacterial cells, holding the potential to overcome antimicrobial resistance and biofilm formation and constituting a promising solution for the treatment of potential fatal multidrug-resistant bacterial infections, such as methicillin resistant Staphylococcus aureus. In this review, we aim to address the applicability of antibiotic encapsulated liposomes as an effective therapeutic strategy for bacterial infections.

scholarly journals Insulin-like growth factor-1 level is a poor diagnostic indicator of growth hormone deficiency

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Hideyuki Iwayama ◽  
Sachiko Kitagawa ◽  
Jyun Sada ◽  
Ryosuke Miyamoto ◽  
Tomohito Hayakawa ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Growth Factor ◽  
Diagnostic Accuracy ◽  
Growth Hormone Deficiency ◽  
Screening Test ◽  
Characteristic Curve ◽  
Hormone Deficiency ◽  
Height Velocity ◽  
Insulin Like Growth Factor ◽  
Predictive Values

AbstractWe evaluated the diagnostic accuracy of insulin-like growth factor-1 (IGF-1) for screening growth hormone deficiency (GHD) to determine the usefulness of IGF-1 as a screening test. Among 298 consecutive children who had short stature or decreased height velocity, we measured IGF-1 levels and performed growth hormone (GH) secretion test using clonidine, arginine, and, in cases with different results of the two tests, L-dopa. Patients with congenital abnormalities were excluded. GHD was defined as peak GH ≤ 6.0 ng/mL in the two tests. We identified 60 and 238 patients with and without GHD, respectively. The mean IGF-1 standard deviation (SD) was not significantly different between the GHD and non-GHD groups (p = 0.23). Receiver operating characteristic curve analysis demonstrated the best diagnostic accuracy at an IGF-1 cutoff of − 1.493 SD, with 0.685 sensitivity, 0.417 specificity, 0.25 positive and 0.823 negative predictive values, and 0.517 area under the curve. Correlation analysis revealed that none of the items of patients’ characteristics increased the diagnostic power of IGF-1. IGF-1 level had poor diagnostic accuracy as a screening test for GHD. Therefore, IGF-1 should not be used alone for GHD screening. A predictive biomarker for GHD should be developed in the future.

scholarly journals Recent Development of Rapid Antimicrobial Susceptibility Testing Methods through Metabolic Profiling of Bacteria

Antibiotics ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 311
Author(s):  
Chen Chen ◽  
Weili Hong
Keyword(s):  
Antimicrobial Susceptibility ◽  
Metabolic Profiling ◽  
Bacterial Infections ◽  
Susceptibility Testing ◽  
Antimicrobial Susceptibility Testing ◽  
Bacterial Metabolism ◽  
Resistant Bacteria ◽  
Bacterial Cells ◽  
Inappropriate Use ◽  
Key Factor

Due to the inappropriate use and overuse of antibiotics, the emergence and spread of antibiotic-resistant bacteria are increasing and have become a major threat to human health. A key factor in the treatment of bacterial infections and slowing down the emergence of antibiotic resistance is to perform antimicrobial susceptibility testing (AST) of infecting bacteria rapidly to prescribe appropriate drugs and reduce the use of broad-spectrum antibiotics. Current phenotypic AST methods based on the detection of bacterial growth are generally reliable but are too slow. There is an urgent need for new methods that can perform AST rapidly. Bacterial metabolism is a fast process, as bacterial cells double about every 20 to 30 min for fast-growing species. Moreover, bacterial metabolism has shown to be related to drug resistance, so a comparison of differences in microbial metabolic processes in the presence or absence of antimicrobials provides an alternative approach to traditional culture for faster AST. In this review, we summarize recent developments in rapid AST methods through metabolic profiling of bacteria under antibiotic treatment.

scholarly journals Duplex Real-Time PCR Assay for Rapid Detection of Ampicillin-Resistant Enterococcus faecium

2004 ◽  
Vol 48 (2) ◽  
pp. 556-560 ◽  
Author(s):  
Stein Christian Mohn ◽  
Arve Ulvik ◽  
Roland Jureen ◽  
Rob J. L. Willems ◽  
Janetta Top ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Enterococcus Faecium ◽  
Rapid Detection ◽  
Resistant Bacteria ◽  
Pcr Assay ◽  
Culture Methods ◽  
Accurate Identification ◽  
Antibiotic Resistant ◽  
Highly Correlated

ABSTRACT Rapid and accurate identification of carriers of resistant microorganisms is an important aspect of efficient infection control in hospitals. Traditional identification methods of antibiotic-resistant bacteria usually take at least 3 to 4 days after sampling. A duplex real-time PCR assay was developed for rapid detection of ampicillin-resistant Enterococcus faecium (ARE). Primers and probes that are used in this assay specifically detected the d-Ala-d-Ala ligase gene of E. faecium and the modified penicillin-binding protein 5 gene (pbp5) carrying the Glu-to-Val substitution at position 629 (Val-629) in a set of 129 tested E. faecium strains with known pbp5 sequence. Presence of the Val-629 in the strain set from 11 different countries was highly correlated with ampicillin resistance. In a screening of hospitalized patients, the real-time PCR assay yielded a sensitivity and a specificity for the detection of ARE colonization of 95% and 100%, respectively. The results were obtained 4 h after samples were harvested from overnight broth of rectal swab samples, identifying both species and the resistance marker mutation in pbp5. This novel assay reliably identifies ARE 2 to 3 days more quickly than traditional culture methods, thereby increasing laboratory throughput, making it useful for rectal screening of ARE. The assay demonstrates the advantages of real-time PCR for detection of nosocomial pathogens.

scholarly journals Studies on genes expression pattern of antioxidant enzymes and enzymes involved into the genetic information implementation in E.coli cells due to the antibiotic resistance against apramycin and cefatoxime

2020 ◽  
Vol 17 ◽  
pp. 00103
Author(s):  
Oleg Fomenko ◽  
Evgeny Mikhailov ◽  
Nadezhda Pasko ◽  
Svetlana Grin ◽  
Andrey Koshchaev ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
Antioxidant Enzymes ◽  
Genetic Information ◽  
Bacterial Resistance ◽  
Resistant Bacteria ◽  
Control Group ◽  
Bacterial Cells ◽  
E Coli ◽  
Antimicrobial Substances ◽  
Dna And Rna

The emergence of antibiotic-resistant bacteria is considered a serious problem. The resistance of bacteria against antimicrobial substances becomes important in the repair systems for damage to DNA and RNA molecules. The role of the antioxidant system in the development of bacterial resistance against antibiotics is not yet practically studied. The article studied the expression regulation of the genes of antioxidant enzymes and enzymes involved in the genetic information in E. coli cells with the antibiotic resistance against apramycin and cefatoxime. The study was conducted on bacterial cells resistant against these two antibiotics. The genes blaOXA-1, blaSHV, blaTEM, mdtK, aadA1, aadA2, sat, strA, blaCTX, blaPER-2, tnpA, tnpR, intC1 and intC1c were identified in bacterial cell case. This indicates the presence of plasmids in bacteria with these genes, which provide bacterial resistance to apramycin and cefatoxime. It was established that during the formation of cefotaxime resistance, there was a sharp increase in the expression of the Cu, Zn superoxide dismutase gene: in comparison with the control group, the representation of its transcripts increased 141.04 times for cefotoxime and 155.42 times for apramycin. It has been established that during the formation of resistance to the studied antibiotics in E. coli, an increase in the expression of the end4 and end3 genes is observed. There is tendency toward an increase in the number of transcripts of the pol3E gene observed in the formation of resistance against cefotaxime and apromycin.

scholarly journals Early Prognostic Factors for the Progress of Preeclampsia – Our Experience in the Period 2010-2011

2016 ◽  
Vol 4 (3) ◽  
pp. 420-422 ◽  
Author(s):  
Mariya Angelova Angelova ◽  
Ivan Todorov ◽  
Emil Kovachev
Keyword(s):  
Pregnant Women ◽  
Prognostic Value ◽  
Protein A ◽  
Low Doses ◽  
Easy Method ◽  
Uterine Arteries ◽  
Non Invasive ◽  
Placental Blood ◽  
Single Child ◽  
Placental Blood Flow

AIM: To determine the prognostic value of the low Pregnancy-associated plasma protein A (PAPP-A) levels in the early stages of pregnancy (11–13 weeks GA) independently and in combination with a Doppler test of the uterine arteries during the second half of pregnancy (22–23 weeks GA).MATERIAL AND METHODS: The study covered the period 2010–2011 and included 106 pregnant women, aged 35–40, with a single child pregnancy. The research excluded pregnant women with anomalies of the fetus, smokers and women taking prophylactically low doses of aspirin.RESULTS: Thirty-six pregnant women had PAPP-A level below 0.4 MoM, whereas 20 of them developed preeclampsia and 7 – early preeclampsia. The combination of the low PAPP-A values and the abnormal Doppler test of the uterine arteries is with a considerably better prognostic value in regards to the risk of developing preeclampsia.CONCLUSION: The Doppler test is a non-invasive, quick and easy method for assessment of the uterine-placental blood flow.

scholarly journals Prevalence of Methicillin Resistant and Virulence Determinants in Clinical Isolates of Staphylococcus aureus

2018 ◽  
Vol 10 (1) ◽  
pp. 108-115
Author(s):  
Manjunath Chavadi ◽  
Rahul Narasanna ◽  
Ashajyothi Chavan ◽  
Ajay Kumar Oli ◽  
Chandrakanth Kelmani. R
Keyword(s):  
Staphylococcus Aureus ◽  
Cell Morphology ◽  
Methicillin Resistance ◽  
Protein A ◽  
Clinical Isolates ◽  
Clinical Samples ◽  
Virulence Determinants ◽  
Methicillin Resistant ◽  
Alternative Medicines ◽  
Mrsa Strains

Introduction:Methicillin-resistantStaphylococcus aureus(MRSA) is the major threat that is a result of the uncontrolled use of antibiotics causing a huge loss in health, so understanding their prevalence is necessary as a public health measure.Objective:The aim of this study was to determine the prevalence of methicillin-resistant MRSA and virulence determinant among associatedS. aureusfrom the clinical samples obtained from various hospital and health care centers of the Gulbarga region in India.Materials and Methods:All the collected samples were subjected for the screening ofS. aureusand were further characterized by conventional and molecular methods including their antibiotic profiling. Further, the response of methicillin antibiotic on cell morphology was studied using scanning electron microscopy.Results:A total 126S. aureuswas isolated from the clinical samples which showed, 100% resistant to penicillin, 55.5% to oxacillin, 75.3% to ampicillin, 70.6% to streptomycin, 66.6% to gentamicin, 8.7% to vancomycin and 6.3% to teicoplanin. The selected MRSA strains were found to possessmecA(gene coding for penicillin-binding protein 2A) andfemA(factor essential for methicillin resistance)genetic determinants in their genome with virulence determinants such as Coagulase (coa) and the X region of the protein A (spa)gene. Further, the methicillin response in resistantS. aureusshowed to be enlarged and malformed on cell morphology.Conclusion:The molecular typing of clinical isolates ofS. aureusin this study was highly virulent and also resistant to methicillin; this will assist health professionals to control, exploration of alternative medicines and new approaches to combat Staphylococcal infections more efficiently by using targeted therapy.

EPISIOTOMY INFECTIONS IN THE PUERPERIUM AND ANTIMICROBIAL RESISTANCE OF RESPONSIBLE PATHOGENS IN UKRAINE

2020 ◽  
Vol 73 (11) ◽  
pp. 2325-2331
Author(s):  
Aidyn G. Salmanov ◽  
Taras G. Voitok ◽  
Igor V. Maidannyk ◽  
Serhiy Yu. Vdovychenko ◽  
Olena О. Chorna ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Antimicrobial Resistance ◽  
Vaginal Delivery ◽  
Prevalence Rate ◽  
Methicillin Resistance ◽  
Carbapenem Resistance ◽  
Resistant Bacteria ◽  
Esbl Production ◽  
Multicenter Cohort Study ◽  
Klebsiella Spp

1 2 ABSTRACT The aim: To obtain the first estimates of the current prevalence rate of episiotomy infections in the puerperium and antimicrobial resistance of responsible pathogens in Ukraine. Materials and methods: We performed a retrospective multicenter cohort study was based on surveillance data. The study population consisted of all women who had a vaginal delivery in 7 Regional Women’s Hospitals of Ukraine. Definitions of episiotomy infections were used from the Centers for Disease Control and Prevention’s National Healthcare Safety Network (CDC/NHSN). Results: Total 35.6% women after vaginal delivery had episiotomy done. The prevalence rate of episiotomy infections was 17.7%. The predominant pathogens were: Escherichia coli (49.2%), Enterobacter spp. (11.1%), Streptococcus spp. (9.1%), Enterococcus faecalis (6.5%), Klebsiella spp. (8.1%), followed by Pseudomonas aeruginosa (4.7%), Staphylococcus aureus (4.2%), Proteus spp.(2.9%) and Staphylococcus epidermidis (2.8%). The overall proportion of methicillin-resistance was observed in 17.3% of Staphylococcus aureus (MRSA). Vancomycin resistance was observed in 6.8% of isolated enterococci. Carbapenem resistance was identified in 8% of P.aeruginosa isolates. Resistance to third-generation cephalosporins was observed in 15.2% Klebsiella spp. and E.coli 16.4% isolates. The overall proportion of extended spectrum beta-lactamases (ESBL) production among Enterobacteriaceae was 26.4%. The prevalence of ESBL production among E. coli isolates was significantly higher than in K. pneumoniae (31.4%, vs 12.5%). Conclusions: Episiotomy infections in the puerperium are common in Ukraine and most of these infections caused by antibiotic-resistant bacteria. Optimizing the management and empirical antimicrobial therapy may reduce the burden of episiotomy infections, but prevention is the key element.

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