scholarly journals Rapid fluoroquinolone resistance detection in Pseudomonas aeruginosa using mismatch amplification mutation assay-based real-time PCR

2021 ◽  
Author(s):  
Danielle E. Madden ◽  
Kate L. McCarthy ◽  
Scott C. Bell ◽  
Olusola Olagoke ◽  
Timothy Baird ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Patient Outcomes ◽  
Health Concern ◽  
Fluoroquinolone Resistance ◽  
Clinical Implementation ◽  
Ciprofloxacin Resistance ◽  
Mutation Assay ◽  
Improve Patient ◽  
Timely Treatment ◽  
Mismatch Amplification Mutation Assay

Antimicrobial resistance (AMR) is an ever-increasing global health concern. Here, we developed two SYBR Green-based mismatch amplification mutation assays (SYBR-MAMAs) targeting GyrA T83I, D87N, D87Y, and D87H, the predominant causes of fluoroquinolone AMR in Pseudomonas aeruginosa. SYBR-MAMAs were tested on 85 isolates, 45 with intermediate/full ciprofloxacin resistance. Assays identified the correct phenotype in 84% intermediate/full resistant and 100% sensitive strains. Clinical implementation of our rapid SYBR-MAMAs will permit timely treatment alterations to improve patient outcomes.

Clinical Utilization of Pharmacogenetics in Psychiatry – Perspectives of Pharmacists, Genetic Counselors, Implementation Science, Clinicians, and Industry

2019 ◽  
Vol 53 (04) ◽  
pp. 162-173 ◽  
Author(s):  
Lisa Brown ◽  
Seenae Eum ◽  
Susanne B. Haga ◽  
Jeffrey R. Strawn ◽  
Heather Zierhut
Keyword(s):  
Patient Outcomes ◽  
Decision Support Tool ◽  
Genetic Counselors ◽  
Clinical Implementation ◽  
Support Tool ◽  
Clinical Utilization ◽  
Medication Side ◽  
Patients At Risk ◽  
Medication Selection ◽  
Improve Patient

Abstract Introduction The use of pharmacogenomic (PGx) testing to guide decisions and improve patient outcomes has increased in recent years. PGx testing represents a decision support tool that may inform dosing, increase the likelihood of treatment response, and identify patients at risk for medication side effects. Methods This is a narrative review of utilization of PGx testing in psychiatry from stakeholders including, pharmacists, genetic counselors, implementation scientists, industry, and clinicians. Results While many limitations exist to streamline use of PGx testing in psychiatry, various stakeholders are crucial to clinical implementation. Discussion PGx testing can assist in medication selection and improve patient outcomes; however, more data are needed to understand when and how to incorporate PGx testing into psychiatric practice.

scholarly journals Ciprofloxacin Resistance in Campylobacter jejuniIsolates: Detection of gyrA Resistance Mutations by Mismatch Amplification Mutation Assay PCR and DNA Sequence Analysis

1999 ◽  
Vol 37 (10) ◽  
pp. 3276-3280 ◽  
Author(s):  
Gerald Zirnstein ◽  
Yu Li ◽  
Bala Swaminathan ◽  
Frederick Angulo
Keyword(s):  
Sequence Analysis ◽  
Dna Sequence ◽  
Campylobacter Jejuni ◽  
Dna Sequence Analysis ◽  
Quinolone Resistance ◽  
Resistance Mutations ◽  
Ciprofloxacin Resistance ◽  
Mutation Assay ◽  
Mismatch Amplification Mutation Assay

The gyrA quinolone resistance determining region was sequenced from 13 ciprofloxacin-resistant and 20 ciprofloxacin-susceptible Campylobacter jejuni isolates. All isolates resistant to ciprofloxacin had Thr-86-to-Ile mutations, a mutation frequently associated with the acquisition of resistance to fluoroquinolones. A mismatch amplification mutation assay (MAMA) PCR protocol was developed that detects this gyrA mutation in quinolone-resistant isolates. The MAMA PCR provides a means for routine detection of the gyrA mutation without the need for sequencing the gyrA gene.

scholarly journals Phenotypic and genotypic examination of antimicrobial resistance in thermophilic Campylobacter species isolated from poultry in Turkey

2018 ◽  
Vol 62 (4) ◽  
pp. 463-468 ◽  
Author(s):  
Mehmet Cemal Adiguzel ◽  
Belgi Diren Sigirci ◽  
Baran Celik ◽  
Beren Basaran Kahraman ◽  
Kemal Metiner ◽  
...  
Keyword(s):  
Nalidixic Acid ◽  
Acid Resistance ◽  
Thermophilic Campylobacter ◽  
Intestinal Contents ◽  
Ciprofloxacin Resistance ◽  
Campylobacter Species ◽  
Rearing Methods ◽  
Mutation Assay ◽  
Nalidixic Acid Resistance ◽  
Mismatch Amplification Mutation Assay

AbstractIntroduction:The study aimed to isolate thermophilicCampylobacterfrom chickens raised three rearing methods, determine its antimicrobial susceptibilities, and examine resistance-related genes by PCR.Material and Methods:Cloacal swabs or intestinal contents were taken in Istanbul, Sakarya, and Izmir provinces. Chickens were from small village-based family-run businesses (n = 70), organically raised (n = 71), and conventionally raised broilers (n = 79). The samples were cultured on modified charcoal cefoperazone desoxycholate (mCCD) agar. Suspect isolates were identified with multiplex PCR (mPCR). As per EUCAST standards, MIC values were derived by broth microdilution for tetracycline, ciprofloxacin, nalidixic acid, kanamycin, gentamicin, and erythromycin in isolates ofC. jejuni(n = 98) andC. coli(n = 83).Results:InC. jejuni, 78.6% tetracycline, 87.8% ciprofloxacin, and 81.6% nalidixic acid resistance was detected, but none was to kanamycin, gentamicin, or erythromycin. InC. coli, 98.8% ciprofloxacin and 63.9% nalidixic acid resistance was detected, whereas resistance to nonquinolones was not observed. C257T (Thr-86-Ile) mutation in thegyrAgene of all phenotypically quinolone-resistant isolates was detected through a mismatch amplification mutation assay PCR (MAMA-PCR). It emerged that all isolates bore thetet(O) resistance gene.Conclusion:Common tetracycline, nalidixic acid, and ciprofloxacin resistance exists inCampylobacterisolated from chickens raised three rearing methods.

scholarly journals Adjunctive S100A8/A9 Immunomodulation Hinders Ciprofloxacin Resistance in Pseudomonas aeruginosa in a Murine Biofilm Wound Model

2021 ◽  
Vol 11 ◽  
Author(s):  
Anne S. Laulund ◽  
Franziska Schwartz ◽  
Hannah Trøstrup ◽  
Kim Thomsen ◽  
Lars Christophersen ◽  
...  
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Chronic Wounds ◽  
Treated Group ◽  
Fluoroquinolone Resistance ◽  
Resistance Pattern ◽  
Adjuvant Effect ◽  
Wound Model ◽  
Inflammatory Proteins ◽  
Ciprofloxacin Resistance

ObjectivePseudomonas aeruginosa is known to contribute to the pathogenesis of chronic wounds by biofilm-establishment with increased tolerance to host response and antibiotics. The neutrophil-factor S100A8/A9 has a promising adjuvant effect when combined with ciprofloxacin, measured by quantitative bacteriology, and increased anti- and lowered pro-inflammatory proteins. We speculated whether a S100A8/A9 supplement could prevent ciprofloxacin resistance in infected wounds.MethodFull-thickness 2.9cm2-necrosis was inflicted on 32 mice. On day 4, P.aeruginosa in seaweed alginate was injected sub-eschar to mimic a mono-pathogenic biofilm. Mice were randomized to receive ciprofloxacin and S100A8/A9 (n=14), ciprofloxacin (n=12) or saline (n=6). Half of the mice in each group were euthanized day 6 and the remaining day 10 post-infection. Mice were treated until sacrifice. Primary endpoint was the appearance of ciprofloxacin resistant P.aeruginosa. The study was further evaluated by genetic characterization of resistance, means of quantitative bacteriology, wound-size and cytokine-production.ResultsThree mice receiving ciprofloxacin monotherapy developed resistance after 14 days. None of the mice receiving combination therapy changed resistance pattern. Sequencing of fluoroquinolone-resistance determining regions in the ciprofloxacin resistant isolates identified two high-resistant strains mutated in gyrA C248T (MIC>32µg/ml) and a gyr B mutation was found in the sample with low level resistance (MIC=3µg/ml). Bacterial densities in wounds were lower in the dual treated group compared to the placebo group on both termination days.ConclusionThis study supports the ciprofloxacin augmenting effect and indicates a protective effect in terms of hindered ciprofloxacin resistance of adjuvant S100A8/A9 in P.aeruginosa biofilm infected chronic wounds.

Combined Approaches to the Skull Base for Intracranial Extension of Tumors via Perineural Spread can Improve Patient Outcomes

2016 ◽  
Vol 77 (S 01) ◽  
Author(s):  
Sheri Palejwala ◽  
Jonnae Barry ◽  
Crystal Rodriguez ◽  
Chandni Parikh ◽  
Stephen Goldstein ◽  
...  
Keyword(s):  
Skull Base ◽  
Patient Outcomes ◽  
Intracranial Extension ◽  
Perineural Spread ◽  
Improve Patient

Microporous Polysaccharide Hemosphere Absorbable Hemostat (AristaAH®) in Re-Operative Cardiac Surgical Procedures

2012 ◽  
Vol 9 (2) ◽  
pp. 96-98
Author(s):  
Brian A Bruckner ◽  
Matthias Loebe
Keyword(s):  
Patient Outcomes ◽  
Surgical Procedures ◽  
Systematic Approach ◽  
Surgical Intervention ◽  
Transfusion Requirement ◽  
Cardiac Surgical Procedures ◽  
Platelet Inhibitors ◽  
Hemostatic Agents ◽  
Surgical Field ◽  
Improve Patient

Patients undergoing re-operative cardiac surgical procedures present a great challenge with regard to obtaining hemostasis in the surgical field. Adhesions are ever-present and these patients are often on oral anti-coagulants and platelet inhibitors. As part of a well-planned surgical intervention, a systematic approach to hemostasis should be employed to decrease blood transfusion requirement and improve patient outcomes. Topical hemostatic agents can be a great help to the surgeon in achieving surgical field hemostasis and are increasingly being employed. Our approach, to these difficult patients, includes the systematic and planned use of AristaAH, which is a novel hemostatic agent whose use has proven safe and efficacious in our patient population.

Bioactivity of Phenolic Compounds Extracted from Strawberry against Formation of Pseudomonas aeruginosa Biofilm

2019 ◽  
Vol 10 (01) ◽  
Author(s):  
Baydaa Hussein ◽  
Zainab A. Aldhaher ◽  
Shahrazad Najem Abdu-Allah ◽  
Adel Hamdan
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Phenolic Compounds ◽  
Biofilm Formation ◽  
Opportunistic Infections ◽  
Formation Rate ◽  
Hydrocarbon Chain ◽  
Health Concern ◽  
Gas Chromatography Mass Spectrometry ◽  
Phenolic Contents

Background: Biofilm is a bacterial way of life prevalent in the world of microbes; in addition to that it is a source of alarm in the field of health concern. Pseudomonas aeruginosa is a pathogenic bacterium responsible for all opportunistic infections such as chronic and severe. Aim of this study: This paper aims to provide an overview of the promotion of isolates to produce a biofilm in vitro under special circumstances, to expose certain antibiotics to produce phenotypic evaluation of biofilm bacteria. Methods and Materials: Three diverse ways were used to inhibited biofilm formation of P.aeruginosa by effect of phenolic compounds extracts from strawberries. Isolates produced biofilm on agar MacConkey under certain circumstances. Results: The results showed that all isolates were resistant to antibiotics except sensitive to azithromycin (AZM, 15μg), and in this study was conducted on three ways to detect the biofilm produced, has been detected by the biofilm like Tissue culture plate (TCP), Tube method (TM), Congo Red Agar (CRA). These methods gave a clear result of these isolates under study. Active compounds were analyzed in both extracts by Gas Chromatography-mass Spectrometry which indicate High molecular weight compound with a long hydrocarbon chain. Conclusion: Phenolic compounds could behave as bioactive material and can be useful to be used in pharmaceutical synthesis. Phenolic contents which found in leaves and fruits extracts of strawberries shows antibacterial activity against all strains tested by the ability to reduce the production of biofilm formation rate.

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