1227. Plazomicin Susceptibility Testing using ETEST® MIC for Enterobacterales

Abstract Background Plazomicin (PLZ) approved by FDA in June of 2018, is an aminoglycoside class antibacterial indicated for the treatment of adults with complicated urinary tract infections (cUTI) including pyelonephritis caused by Enterobacterales. It is used in patients who have limited or no alternative treatment options, e.g. CRE and MDRO patients. The drug has bactericidal activity, it is active against organisms producing ESBL, Carbapenemase and aminoglycoside-modifying enzymes. The purpose of this study was to compare ETEST® PLZ bioMérieux to the broth microdilution reference method (BMD) for Citrobacter freundii, Enterobacter cloacae, Escherichia coli, Klebsiella oxytoca and pneumoniae, Morganella morganii, Providencia stuartii, Proteus mirabilis and vulgaris and Serratia marcescens isolates. Methods A total of 598 isolates were tested by ETEST® (PLZ) and BMD at four clinical trial sites. Isolates were subcultured on tryptic soy or Columbia agar plates supplemented with 5% sheep blood. Suspensions of the isolates were prepared in 0.85% saline, which were used to inoculate BMD and Mueller Hinton agar for ETEST®. Results were read after 16-20 hours incubation at 35°C +2°C in ambient air. QC organisms were tested with each run following CLSI QC guidelines. Results were analyzed using FDA breakpoints for PLZ (Susceptible <2 µg/mL, Intermediate 4 µg/mL, Resistant >8 µg/mL). Performance was evaluated using FDA performance criteria, EA and CA (≥ 90%), major error rate (≤3.0%) and very major error rate (≤2.0%). Results Conclusion ETEST® PLZ clinical performance met the FDA acceptance criteria and was found useful for determining Plazomicin MIC of Enterobacterales, including ESBL, CRE (MBL, KPC, Oxa-48), high level AmpC and aminoglycoside resistant strains. Percent susceptibility of Plazomicin is at 80% among the 598 isolates tested, the mode MIC is 0.5 ug/ml as Susceptible. Disclosures Tom Armstrong, BS, bioMérieux (Employee) Laurine Blanchard, PhD, bioMérieux (Employee) Michael Kresken, PhD, bioMérieux (Scientific Research Study Investigator, Research Grant or Support) Gilles Zambardi, biomerieux (Employee) Marion Pompilio, BioMérieux (Employee)

Download Full-text

1222. Fosfomycin Susceptibility Testing using the new ETEST®FO

Open Forum Infectious Diseases ◽

10.1093/ofid/ofaa439.1407 ◽

2020 ◽

Vol 7 (Supplement_1) ◽

pp. S632-S632

Author(s):

Diane Halimi ◽

Marion Pompilio ◽

Sandrine Fontaine ◽

Roland Martelin ◽

Christine Franceschi ◽

...

Keyword(s):

Error Rate ◽

Reference Method ◽

Ambient Air ◽

Performance Criteria ◽

Study Phase ◽

Major Error ◽

E Coli ◽

Wide Range ◽

Agar Dilution ◽

Tract Infections

Abstract Background Fosfomycin (FO) is a bactericidal antibiotic with a broad spectrum of activity against a wide range of Gram-positive and Gram-negative bacteria. Oral FO is mainly used in the treatment of urinary tract infections, particularly those caused by E. coli and E. faecalis. In order to determine MICs to FO, the ETEST® FM is already available but the reading can be difficult especially with E. coli. To resolve this issue, a new ETEST® with FO, called ETEST® FO, has been developed (not FDA cleared, yet). The purpose of this study is to compare this new strip to the agar dilution reference method (AD) on a panel of E. coli and E. faecalis. Methods A total of 39 isolates comprising 20 E. coli (ESBL or CPE) and 19 E. faecalis (VRE or VSE) were tested by ETEST® FO and Agar dilution. The isolates were sub-cultured on Columbia agar plates supplemented with 5% sheep blood before testing. After incubation, suspensions of the isolates were prepared in 0.85% saline. These suspensions were used to inoculate both AD and ETEST® plates. Results were read after 16-20 hours incubation at 35°C +2°C in ambient air. Following CLSI QC guideline, 4 QC organisms were tested. Results were analyzed using the FDA/CLSI breakpoints for FO (S < 64µg/mL, I=128 µg/mL, R> 256 µg/mL). Performance was evaluated using FDA performance criteria, essential agreement (EA, ≥ 90%), category agreement (CA, ≥ 90%), major error rate (ME, ≤3.0%) and very major error rate (VME, ≤2.0%). Results All the QC strains MICs were within the CLSI ranges. For the panel results, see the table below: Performance for ETEST® FO on E. coli and E. faecalis Conclusion This first and preliminary study shows that ETEST® FO can potentially meet the FDA acceptance criteria and could be a valuable tool for determining FO MIC for E. coli harboring various resistance mechanisms and E. faecalis including VRE. Moreover, in comparison with the current ETEST FM strip, this new strip brings a real reading improvement and resolve the issue for E. coli. The clinical study phase will determine the product’s performance. Disclosures Marion Pompilio, BioMérieux (Employee) Gilles Zambardi, biomerieux (Employee)

Download Full-text

Multicenter Evaluation of Ceftazidime-Avibactam Susceptibility Testing of Enterobacterales and Pseudomonas aeruginosa on the VITEK® 2 System

Journal of Clinical Microbiology ◽

10.1128/jcm.01870-20 ◽

2020 ◽

Author(s):

Romney Humphries ◽

Shelley Campeau ◽

Thomas E. Davis ◽

Kristin J. Nagaro ◽

Vincent J. LaBombardi ◽

...

Keyword(s):

Pseudomonas Aeruginosa ◽

Error Rate ◽

Susceptibility Testing ◽

Reference Method ◽

Error Rates ◽

Performance Criteria ◽

Test Results ◽

Major Error ◽

Overall Performance ◽

Vitek 2

In this multisite study, VITEK® 2 AST-Gram-Negative Ceftazidime-Avibactam (CZA) test results for 1073 isolates (866 Enterobacterales and 207 Pseudomonas aeruginosa) were compared to the Clinical & Laboratory Standards Institute (CLSI) broth microdilution (BMD) reference method. The results were analyzed for essential agreement (EA), category agreement (CA), major error rates, and very major error rates following FDA/ISO performance criteria using the FDA-recognized CLSI/EUCAST breakpoints (S ≤8/4 μg/ml and R ≥16/4 μg/ml). The overall EA was 94.5% (1014/1073) and CA was 98.7% (1059/1073). No very major errors were reported. The major error rate was 1.4% (14/998). Out of 14 major errors, 9 were within EA. Based on the EA and lack of an intermediate category for CZA, the adjusted major error rate for FDA criteria was 0.5% (5/998). The performance for ISO criteria after error resolutions included EA 94.5% (1014/1073), CA 98.9% (1061/1073), major error 1.2% (12/998), and no very major error. Vitek 2 met the ISO and FDA criteria of ≥95% reproducibility and ≥95% quality control (QC) results within acceptable ranges for QC organisms. Vitek 2 overall performance for Enterobacterales and P. aeruginosa met or exceeded the FDA and ISO performance criteria and thus is a reliable alternative to BMD reference method for routine CZA susceptibility testing.

Download Full-text

Antibiotic susceptibility profile of extended spectrum β-lactamase producing Escherichia coli, Klebsiella pneumoniae and Klebsiella oxytoca from Urinary tract infections

Research Journal of Pharmacy and Technology ◽

10.52711/0974-360x.2021.00768 ◽

2021 ◽

pp. 4425-4428

Author(s):

M. Sharmal Kumar ◽

Arunagirinathan N. ◽

Ravikumar M.

Keyword(s):

Escherichia Coli ◽

Urinary Tract ◽

Urinary Tract Infections ◽

Antibiotic Susceptibility ◽

Klebsiella Oxytoca ◽

Bacterial Pathogen ◽

Resistant Bacteria ◽

Extended Spectrum ◽

High Level ◽

Tract Infections

The aim of this study was to analyze the extended spectrum β-lactamases (ESBLs) production and antibiotic susceptibility profile of urinary tract infected bacterial pathogens such as Escherichia coli and Klebsiella spp. A total of 143 Gram-negative bacteria were isolated from people suffering from urinary tract infections (UTIs) were included in this study. Among them, Escherichia coli (75%) were the predominantly isolated bacterial pathogen followed by Klebsiella oxytoca (14.6%) and K. pneumoniae (10.4%). Approximately 65% (n=93) of isolates were positive for ESBL production and E.coli was found to be the highest ESBL producer (67.6%) followed by K. oxytoca (57.1%) and K. pneumoniae (53.3%). E. coli showed high level of 86.1% resistance to cefotaxime and cefuroxime and 100% sensitive to imipenem and meropenem, whereas, K. oxytoca showed high level of 90.5% resistance to cefuroxime and 100% sensitive to amikacin, imipenem and meropenem. Similarly, K. pneumoniae showed high level of 73.3% resistance to nitrofurantoin and 93.3% sensitive to imipenem. This study reveals that majority of UTIs caused bacteria are ESBL producing multidrug-resistant bacteria and showing broad spectrum antibiotic resistance profile.

Download Full-text

Assessment of the performance of blood glucose monitoring systems for monitoring dysglycaemia in neonatal patients

BMJ Paediatrics Open ◽

10.1136/bmjpo-2018-000339 ◽

2018 ◽

Vol 2 (1) ◽

pp. e000339 ◽

Cited By ~ 3

Author(s):

Yin Ba ◽

Jin Xu ◽

Lin Yuan ◽

Haiyan Zhu ◽

Yipei Yang ◽

...

Keyword(s):

Blood Glucose ◽

Clinical Performance ◽

Glucose Monitoring ◽

Reference Method ◽

Performance Criteria ◽

Blood Glucose Monitoring ◽

Monitoring Systems ◽

Clinical Risk ◽

Laboratory Reference ◽

Clinical Risk Assessment

ObjectiveTo validate a three-step protocol that assesses the clinical risk associated with using blood glucose monitoring systems (BGMS) in neonates for the management of dysglycaemia.MethodThe three-step validation approach included confirmation of the accuracy of the reference method using National Institute of Standards and Technology (NIST) glucose standards, assessment of analytical risk performed on whole blood collected from paediatric patients routinely tested for glucose and a clinical risk assessment performed using heel stick capillary samples collected from 147 new-born babies and neonates admitted to intensive care. BGMS glucose measurements were compared with the NIST aligned laboratory reference method.ResultsThe accuracy of the laboratory reference method was confirmed with the NIST standards. Specificity studies demonstrated that the accuracy of one of the BGMS was affected, particularly, in the hypoglycaemic range, by known interference factors including haematocrit, ascorbic acid, lactose, galactose, N-acetylcysteine and glutathione. The accuracy of the other BGMS was unaffected. The clinical performance of this BGMS in neonates met the system accuracy criteria of Clinical and Laboratory Standards Institute (CLSI) POCT 12-A3 standard for evaluating hospital BGMS with 95.1% of glucose measurements within±0.67 mmol/L for samples ≤5.55 mmol/L and 95.6% within±12.5% for samples>5.55 mmol/L.ConclusionsThis three-step validation protocol provides a challenging approach for determining the accuracy and reliability of BGMS for managing dysglycaemia in neonates. StatStrip BGMS achieved analytical and clinical performance criteria confirming its suitability for use in neonates. We advocate that this validation approach should be considered for performance evaluations of both BGMS and continuous glucose monitoring systems going forward.

Download Full-text

Detection of Plasmid-Mediated qnr Genes Among the Clinical Quinolone-Resistant Escherichia coli Strains Isolated in Tehran, Iran

The Open Microbiology Journal ◽

10.2174/1874285801812010248 ◽

2018 ◽

Vol 12 (1) ◽

pp. 248-253 ◽

Cited By ~ 1

Author(s):

Reza Ranjbar ◽

Sajjad S. Tolon ◽

Mehrdad Sami ◽

Reza Golmohammadi

Keyword(s):

Escherichia Coli ◽

Urinary Tract ◽

Nalidixic Acid ◽

Treatment Options ◽

Bacterial Genome ◽

Multidrug Resistant ◽

Documentation System ◽

E Coli ◽

High Level ◽

Tract Infections

Background:Escherichia coliis one of the most important bacterial agents to cause urinary tract infections. Inappropriate and unnecessary administration of antibiotics has led to an increase in the appearance of multidrug-resistantE. coliisolates, limiting treatment options. The increase in a number of resistant strains of bacteria is a major concern of health authorities worldwide.Objective:The purpose of this study was to determine the presence of theqnrgenes amongE. coliisolated from UTIs of patients in Baqiyatallah hospital in Tehran province, Iran.Method:Clinical urine samples of patients with suspected urinary tract infection were collected by standard methods in sterile disposable containers. After analysis of urine, microscopic observations and culture analysis, the bacterial genome was extracted by boiling method. PCR for detection ofqnrgenes includingqnrA,qnrBandqnrSwas done by specific primers, then PCR products were run using gel electrophoresis and visualized by gel documentation system.Results:In the present study among the 95 isolates, 60 strains were resistant to nalidixic acid. PCR showed that 92 strains were positive forqnrS. TheqnrAandqnrBgenes were not found among the clinical isolates.Conclusion:Our finding indicates a high level of resistance against nalidixic acid amongE. coliisolates recovered from the patients with UTI. Also, the high frequency ofqnrSimposes the importance of survey of molecular and genetic analysis of mechanisms of quinolone resistance inE. colistrains.

Download Full-text

Evaluation of an In-House Colistin NP Test for Use in Resource-Limited Settings

Journal of Clinical Microbiology ◽

10.1128/jcm.00501-19 ◽

2019 ◽

Vol 57 (10) ◽

Author(s):

B. Mitton ◽

C. Kingsburgh ◽

M. M. Kock ◽

N. M. Mbelle ◽

K. Strydom

Keyword(s):

Error Rate ◽

Screening Method ◽

Reference Method ◽

Accurate Method ◽

Rapid Screening ◽

Major Error ◽

Colistin Resistance ◽

Screening Assay ◽

Resource Limited Settings ◽

Resource Limited

ABSTRACT Colistin has become increasingly important in the treatment of multidrug-resistant Gram-negative bacteria. Resistance to colistin has emerged globally, necessitating the need for an accurate method to detect colistin resistance. The colistin NP test has shown promise as a rapid screening assay for colistin resistance. This study compared the performance of an in-house-prepared colistin NP test against broth microdilution (BMD) as the gold standard and against Etest (bioMérieux, Marcy l’Etoile, France) as an alternative method. A total of 215 stored Enterobacteriaceae isolates were evaluated, of which 159 were resistant and 56 susceptible to colistin by BMD. The categorical agreement of the colistin NP test with BMD was found to be 98.1%, compared to 87.9% for the Etest. One major error was detected with both the colistin NP test and the Etest. Three very major errors were detected with the colistin NP test compared to 25 with the Etest. This resulted in a major error rate of 1.8% for both the colistin NP test and the Etest and a very major error rate of 1.9% and 15.7% for the colistin NP test and the Etest, respectively. The colistin NP test compared satisfactorily to the BMD reference method in determining colistin susceptibility. The colistin NP test is a rapid, inexpensive screening method for colistin resistance, especially in resource-limited settings.

Download Full-text

Biosimilars Regulation in the United States and FDA Approved Biosimilars from 2015-2018

Applied Clinical Research Clinical Trials and Regulatory Affairs ◽

10.2174/2213476x06666190213141839 ◽

2020 ◽

Vol 7 (1) ◽

pp. 12-29

Author(s):

Vikram ◽

Aakash Deep ◽

Manita ◽

Avtar C. Rana ◽

Monu Yadav ◽

...

Keyword(s):

Price Competition ◽

Clinical Performance ◽

Treatment Options ◽

The United States ◽

Living Cells ◽

Human Beings ◽

Biological Products ◽

Technical Requirements ◽

Manufacturing Techniques ◽

High Level

Background: Biological products are the chemicals in the form of medicines that are prepared from the living cells through highly intricate manufacturing techniques that should be handled and managed under favorable conditions. The regulation of the biosimilar products consists of significant challenges, since they are part of the growing sector of the pharmaceutical industry and normally used by human beings. The regulatory framework and the technical requirements of the US biosimilars program involve a stepwise approach that relies heavily on analytical methods to demonstrate through a “totality of the evidence” that a proposed product is biosimilar to its reference product. By integrating analytical, pharmacological, and clinical data, each of which has limitations, a high level of confidence can be reached regarding clinical performance. The Biologics Price Competition and Innovation Act of 2009 (BPCI Act) was passed as part of the health reform (Affordable Care Act). Objective: The current manuscript will provide the information regarding the regulation of Biosimilars products with the detail of biosimilar USER fees structure and the list of approved biosimilar by FDA from 2015- 2018. Conclusion: Research is continually developing more biological products that will help treat medical conditions or add some innovation to the existing treatment options. Biosimilars and reference products are generated in the living cells and require trained expertise as well as technology for biologics being usually highly effective compared to small molecule drugs. These are usually specific against the respective target, which generally produces lesser side effects and low toxicity. FDA’s regulatory authority for the approval of biologics is under PHS (Public Health Service Act) which are also suggested to regulate under the Federal Food, Drug and Cosmetics Act (FD&C). Biosimilars can help expand access to high-quality treatment options for doctors and patients, as well as reduce costs for the healthcare system.

Download Full-text

Multicenter Evaluation of the BD Phoenix CPO Detect Test for Detection and Classification of Carbapenemase-Producing Organisms in Clinical Isolates

Journal of Clinical Microbiology ◽

10.1128/jcm.01752-19 ◽

2020 ◽

Vol 58 (5) ◽

Author(s):

Vicki Whitley ◽

Susan Kircher ◽

Tracey Gill ◽

Janet A. Hindler ◽

Susan O’Rourke ◽

...

Keyword(s):

Pseudomonas Aeruginosa ◽

Acinetobacter Baumannii ◽

Bacterial Infections ◽

Clinical Performance ◽

Treatment Options ◽

Reference Method ◽

Patient Treatment ◽

High Morbidity ◽

Content Type

ABSTRACT Limited treatment options contribute to high morbidity/mortality rates with carbapenem-resistant, Gram-negative bacterial infections. New approaches for carbapenemase-producing organism (CPO) detection may help inform clinician decision-making on patient treatment and infection control. BD Phoenix CPO detect (CPO detect) detects and classifies carbapenemases in Enterobacterales, Acinetobacter baumannii, and Pseudomonas aeruginosa during susceptibility testing. The clinical performance of CPO detect is reported here. Enterobacterales, Acinetobacter baumannii, and Pseudomonas aeruginosa isolates were evaluated across three sites using CPO detect and a composite reference method (RM); the latter was comprised of the modified carbapenem inactivation method and a MIC screen for ertapenem, imipenem, and meropenem. Multiplex PCR testing was also utilized for Ambler class determination. Positive and negative percentages of agreement (PPA and NPA, respectively) between CPO detect and the RM were determined. The PPA and NPA for Enterobacterales were 98.5% (confidence intervals, 96.6%, 99.4%) and 97.2% (95.8%, 98.2%), respectively. The A. baumannii PPA and NPA, respectively, were 97.1% (90.2%, 99.2%) and 97.1% (89.9%, 99.2%). The P. aeruginosa PPA and NPA, respectively, were 95.9% (88.6%, 98.6%) and 92.3% (86.7%, 95.6%). The PPA values for carbapenemase class designations for all organisms combined and Enterobacterales alone, respectively, were 95.3% (90.2%, 97.8%) and 94.6% (88.8%, 97.5%) for class A, 94.0% (88.7%, 96.6%) and 96.4% (90.0%, 98.8%) for class B, and 95.0% (90.1%, 97.6%) and 99.0% (94.4%, 99.8%) for class D carbapenemases. NPA values for all organisms and Enterobacterales alone ranged from 98.5% to 100%. CPO detect provided accurate detection and classification of CPOs for the majority of isolates of Enterobacterales, Acinetobacter baumannii, and Pseudomonas aeruginosa tested.

Download Full-text

In Vivo Selection of a Chromosomally Encodedβ -Lactamase Variant Conferring Ceftazidime Resistance in Klebsiella oxytoca

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.47.12.3739-3742.2003 ◽

2003 ◽

Vol 47 (12) ◽

pp. 3739-3742 ◽

Cited By ~ 21

Author(s):

Hedi Mammeri ◽

Laurent Poirel ◽

Patrice Nordmann

Keyword(s):

Biochemical Analysis ◽

Klebsiella Oxytoca ◽

Sequencing Analysis ◽

Single Nucleotide ◽

In Vivo Selection ◽

Pcr Products ◽

High Level ◽

Tract Infections ◽

Selection Of

ABSTRACT Klebsiella oxytoca clinical isolate A was recovered from the urine of a 55-year-old man with prostatic and urinary tract infections. This isolate displayed a β-lactam resistance phenotype consistent with overproduction of a chromosomally encoded class Aβ -lactamase and had decreased susceptibilities to allβ -lactams except ceftazidime, cephamycins, and carbapenems. Four weeks after treatment with an antibiotic regimen that included ceftazidime, K. oxytoca isolate B, which had a high level of resistance to ceftazidime, was isolated from the urine of the same patient. Isoelectric focusing analysis of the culture extracts of these isolates gave a pI of 5.4 for both isolates. Cloning experiments with the PCR products of the bla OXY gene resulted in two Escherichia coli DH10B recombinant clones with resistance phenotypes mirroring those of the parental isolates. Sequencing analysis revealed that the blaOXY-2-5 gene from K. oxytoca B had a single nucleotide substitution compared to the sequence of the bla OXY-2 gene from K. oxytoca A, leading to a proline-to-serine substitution at position 167, according to the numbering of Ambler. Biochemical analysis of purified OXY-2-5 showed that it had the ability to hydrolyze ceftazidime. This is the first report of in vivo selection of a K. oxytoca isolate that produced a chromosomally encodedβ -lactamase conferring resistance to ceftazidime.

Download Full-text