scholarly journals Detection of Carbapenem-Resistant Enterobacterales in Simulated Blood Culture in 15 Minutes

Life ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 145
Author(s):  
Daria Baer ◽  
Maya Azrad ◽  
Nora Saleh ◽  
Avi Peretz
Keyword(s):  
Blood Culture ◽  
Medical Center ◽  
Bloodstream Infections ◽  
Resistant Bacteria ◽  
Clinical Microbiology Laboratory ◽  
Antibiotic Resistant ◽  
Culture Bottle ◽  
Carbapenem Resistant ◽  
Life Threatening ◽  
Threatening Situation

Bacteremia leading to sepsis and organ dysfunction is a life-threatening situation, leading to death of up to one fourth of the infected individuals around the world. One major challenge in the treatment of sepsis is the rising prevalence of antibiotic resistant bacteria, such as carbapenem-resistant Enterobacterales (CRE). In recent years, several molecular assays have been developed for the detection of CRE mechanisms, enabling rapid results reporting. We evaluated the performance of the NG-Test CARBA 5 (NG Biotech) kit in detection of CRE in simulated blood cultures. Carbapenemase-producing (CP) CRE isolates (n = 38) and non-carbapenemase CRE (Non-CP) isolates (n = 10), previously identified using the routine methods practiced at the clinical microbiology laboratory of the Baruch Padeh Medical Center, Israel, were used in this analysis. Variable concentrations of the bacterial isolates were added to a suspension composed of human blood and saline, simulating the composition of a blood culture. Samples were then transferred to an anaerobic blood culture bottle and later tested with the NG-Test CARBA 5 (NG Biotech) kit, that identifies the CRE mechanism within 15 min. The NG-Test CARBA 5 kit correctly identified 43 samples (89.5%). The sensitivity and specificity of the kits were 86.8% and 100%, respectively. In conclusion, the NG-Test CARBA 5 kit is a reliable and accessible tool for the rapid diagnosis of CRE bloodstream infections.

Detection of carbapenemase-resistant Enterobacterales in blood culture in 15 minutes

2020 ◽  
Author(s):  
Daria Baer ◽  
Maya Azrad ◽  
Nora Saleh ◽  
Avi Peretz
Keyword(s):  
Blood Culture ◽  
Medical Center ◽  
Bloodstream Infections ◽  
Resistant Bacteria ◽  
Clinical Microbiology Laboratory ◽  
Antibiotic Resistant ◽  
Culture Bottle ◽  
Carbapenem Resistant ◽  
Life Threatening ◽  
Threatening Situation

Abstract Background: Bacteremia leading to sepsis and organ dysfunction is a life-threatening situation. One major challenge in treatment of sepsis is the uprising prevalence of antibiotic resistant bacteria, such as Carbapenem-resistant Enterobacterales (CRE). In recent years several molecular assays have been developed for the detection of CRE mechanisms, enabling rapid results reporting. We evaluated the performance of NG-Test CARBA 5 (NG Biotech) kit for detection of CRE, directly from patients’ blood culture.Methods: Carbapenemase-producing (CP) CRE-positive isolates (n=38) and non-carbapenemase CRE (non-CP) isolates (n=10), previously identified using the routine methods practiced at the clinical microbiology laboratory of the Baruch Padeh Medical Center, Israel were used in this analysis. Variable concentrations of the bacterial isolates were added to a suspension composed of blood unit and saline simulating the composition of a blood culture. Samples were then transferred to an anaerobic blood culture bottle and later tested it with the NG-Test CARBA 5 (NG Biotech) kit, that identifies the CRE mechanism in 15 minutes.Results: The NG-Test CARBA 5 kit correctly identified 43 samples (89.5%). The sensitivity and specificity of the kit were 86.8% and 100%, respectively.Conclusions: The NG-Test CARBA 5 kit is a reliable and accessible tool for the rapid diagnosis of CRE bloodstream infections.

scholarly journals 654. Performance of the T2Resistance Panel in Detecting Antibiotic Resistant Bacteria Directly in Whole Blood, and Implications for Improving Appropriate Therapy of Bloodstream Infections

2021 ◽  
Vol 8 (Supplement_1) ◽  
pp. S429-S429
Author(s):  
Abigail Skeel ◽  
Cornelius J Clancy ◽  
Aaron Lucas ◽  
Kailey L Hughes ◽  
Ryan K Shields ◽  
...  
Keyword(s):  
Blood Culture ◽  
Whole Blood ◽  
Bloodstream Infections ◽  
Resistant Bacteria ◽  
Gram Stain ◽  
Research Support ◽  
Antibiotic Resistant ◽  
Culture Independent ◽  
Appropriate Therapy ◽  
Resistance Markers

Abstract Background Appropriate antibiotic (Ab) therapy of bloodstream infections (BSI) is often delayed by time to blood culture (BC) positivity, species (sp) identification and Ab sensitivity (sensi). The T2Resistance (T2R) Panel is a direct-from-blood (culture-independent) diagnostic that detects 13 genetic markers associated with methicillin-resistant S. aureus (MRSA), vancomycin-resistant Enterococcus (VRE), ESBL- and carbapenemase-producing Enterobacteriaceae (E). We assessed T2R performance in detecting these resistant bacteria in whole blood (WB) and analyzed possible impact on time to appropriate Ab. Methods We performed T2R using WB samples obtained from patients (pts) on the same day as BCs from July 2019-2020. Receipt of appropriate Ab was assessed at time of empiric, Gram stain-directed, MALDI-directed (sp identification) and sensi-directed therapy. T2R results were not available to care teams. Teams were notified of positive BCs. Stewardship optimized Abs based on sensi. Results BC from 103 pts grew 114 bacterial sp: E (n=54; 16 ESBL-, 1 KPC-producer), S. aureus (n=29, 22 MRSA), Enterococcus (n=21, 16 VRE), P. aeruginosa and others (n=10). 12 ESBL-E produced CTX-M 14/15. T2R sensitivity and specificity was 78% and 99%, respectively, compared to sequencing of resistance markers. Sensitivity was excellent for vanA/B, KPC (100% each), and CTX-M14/15 (92%); sensitivity was 58% for mecA/C. T2R detected resistance determinants in 3-7h. Median time to appropriate Ab was 16.3h, which was significantly longer for VRE (25.6h) and ESBL- or KPC-E (50.9h) BSIs than for T2R marker-negative bacteria (6.7h; p=0.04). Pts with VRE or ESBL-/KPC-E BSI were less likely to received appropriate empiric Ab (18% and 30%, respectively) than pts with T2R marker-negative BSI (63%; p=0.02; Fig.1). Median times to achieve ≥80% appropriate Ab therapy of marker-negative, VRE and CTX-M/KPC-E BSIs were 15.5h (after Gram stain), 43.9h (after MALDI) and 63.5h (after sensi), respectively. Antibiotic Therapy Conclusion There was a significant delay in appropriate Ab therapy of BSIs, especially in pts infected with VRE and ESBL/KPC-E. T2R rapidly and accurately detected BSI caused by VRE and ESBL/KPC-E, and has the potential to significantly shorten time to appropriate Ab. Disclosures Cornelius J. Clancy, MD, Merck (Grant/Research Support) Ryan K. Shields, PharmD, MS, Shionogi (Consultant, Research Grant or Support) Minh-Hong Nguyen, MD, Merck (Grant/Research Support)

scholarly journals Prevalence of Antibiotic-Resistant Bacteria ESKAPE among Healthy People Estimated by Monitoring of Municipal Wastewater

Antibiotics ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 495
Author(s):  
Masateru Nishiyama ◽  
Susan Praise ◽  
Keiichi Tsurumaki ◽  
Hiroaki Baba ◽  
Hajime Kanamori ◽  
...  
Keyword(s):  
Antibiotic Resistance ◽  
General Population ◽  
Municipal Wastewater ◽  
Treatment Plant ◽  
Multidrug Resistant ◽  
Resistant Bacteria ◽  
Municipal Wastewater Treatment ◽  
Antibiotic Resistant ◽  
Carbapenem Resistant ◽  
Wastewater Samples

There is increasing attention toward factors that potentially contribute to antibiotic resistance (AR), as well as an interest in exploring the emergence and occurrence of antibiotic resistance bacteria (ARB). We monitored six ARBs that cause hospital outbreaks in wastewater influent to highlight the presence of these ARBs in the general population. We analyzed wastewater samples from a municipal wastewater treatment plant (MWWTP) and hospital wastewater (HW) for six species of ARB: Carbapenem-resistant Enterobacteria (CARBA), extended-spectrum β-lactamase producing Enterobacteria (ESBL), multidrug-resistant Acinetobacter (MDRA), multidrug-resistant Pseudomonas aeruginosa (MDRP), methicillin-resistant Staphylococcus aureus (MRSA), and vancomycin-resistant Enterococci (VRE). We registered a high percentage of ARBs in MWWTP samples (>66%) for all ARBs except for MDRP, indicating a high prevalence in the population. Percentages in HW samples were low (<78%), and no VRE was detected throughout the study. CARBA and ESBL were detected in all wastewater samples, whereas MDRA and MRSA had a high abundance. This result demonstrated the functionality of using raw wastewater at MWWTP to monitor the presence and extent of ARB in healthy populations. This kind of surveillance will contribute to strengthening the efforts toward reducing ARBs through the detection of ARBs to which the general population is exposed.

Aminoglycoside antibiotic resistance conferred by Hpa2 of MDR Acinetobacter baumannii: an unusual adaptation of a common histone acetyltransferase

2019 ◽  
Vol 476 (5) ◽  
pp. 795-808 ◽  
Author(s):  
Jyoti Singh Tomar ◽  
Rama Krishna Peddinti ◽  
Ramakrishna V. Hosur
Keyword(s):  
Acinetobacter Baumannii ◽  
Histone Acetyltransferase ◽  
Hydrophobic Interactions ◽  
Aminoglycoside Antibiotic ◽  
Aminoglycoside Antibiotics ◽  
Titration Calorimetry ◽  
Resistant Bacteria ◽  
Antibiotic Resistant ◽  
Carbapenem Resistant ◽  
Eskape Pathogens

AbstractAntibiotic-resistant bacteria pose the greatest threat to human health. Among the list of such bacteria released by WHO, carbapenem-resistant Acinetobacter baumannii, for which almost no treatment exists, tops the list. A. baumannii is one of the most troublesome ESKAPE pathogens and mechanisms that have facilitated its rise as a successful pathogen are not well studied. Efforts in this direction have resulted in the identification of Hpa2-Ab, an uncharacterized histone acetyltransferase enzyme of GNAT superfamily. Here, we show that Hpa2-Ab confers resistance against aminoglycoside antibiotics using Escherichia coli DH5α strains in which Hpa2 gene is expressed. Resistivity for aminoglycoside antibiotics is demonstrated with the help of CLSI-2010 and KB tests. Isothermal titration calorimetry, MALDI and acetylation assays indicate that conferred resistance is an outcome of evolved antibiotic acetylation capacity in this. Hpa2 is known to acetylate nuclear molecules; however, here it is found to cross its boundary and participate in other functions. An array of biochemical and biophysical techniques were also used to study this protein, which demonstrates that Hpa2-Ab is intrinsically oligomeric in nature, exists primarily as a dimer and its interface is mainly stabilized by hydrophobic interactions. Our work demonstrates an evolved survival strategy by A. baumannii and provides insights into the mechanism that facilitates it to rise as a successful pathogen.

scholarly journals Blurred molecular epidemiological lines between the two dominant methicillin-resistant Staphylococcus aureus clones

2018 ◽  
Author(s):  
Amy C. Dupper ◽  
Mitchell J. Sullivan ◽  
Kieran I. Chacko ◽  
Aaron Mishkin ◽  
Brianne Ciferri ◽  
...  
Keyword(s):  
New York ◽  
Staphylococcus Aureus ◽  
Medical Center ◽  
Data Extraction ◽  
Age Groups ◽  
Bloodstream Infections ◽  
Methicillin Resistant ◽  
Peripheral Intravenous Catheter ◽  
Life Threatening ◽  
Healthcare Associated

AbstractBackgroundMethicillin-resistant Staphylococcus aureus (MRSA) causes life-threatening infections in both community and hospital settings and is a leading cause of healthcare-associated infections (HAIs). We sought to describe the molecular epidemiological landscape of patients with MRSA bloodstream infections (BSIs) at an urban medical center by evaluating the clinical characteristics associated with the two dominant endemic clones.MethodsComprehensive clinical data extraction from the electronic health records of 227 hospitalized patients ≥18 years old with MRSA BSI over a 33-month period in New York City were collected. The descriptive epidemiology and mortality associated with the two dominant clones was compared using logistic regression.ResultsMolecular analysis revealed that 91% of all single-patient MRSA BSIs were due to two equally represented genotypes, clonal complex (CC) 5 (N=117) and CC8 (N=110). MRSA BSIs were associated with a 90-day mortality of 27%. CC8 caused disease more frequently in younger age groups (56 ± 17 vs 67 ± 17 years old; p<0.001) and in non-White race (OR=3.45 95% CI [1.51-7.87]; p=0.003), with few other major distinguishing features. Morbidity and mortality also did not differ significantly between the two clones. CC8 caused BSIs more frequently in the setting of peripheral intravenous catheters (OR=5.96; 95% CI [1.51-23.50]; p=0.01).ConclusionThe clinical features distinguishing dominant MRSA clones continue to converge. The association of CC8 with peripheral intravenous catheter infections underscores the importance of classical community clones causing hospital-onset infections. Ongoing monitoring and analysis of the dynamic epidemiology of this endemic pathogen is crucial to inform management to prevent disease.

scholarly journals Fighting Antibiotic-Resistant Klebsiella pneumoniae with “Sweet” Immune Targets

mBio ◽  
2018 ◽  
Vol 9 (3) ◽  
Author(s):  
Roberto Adamo ◽  
Immaculada Margarit
Keyword(s):  
Klebsiella Pneumoniae ◽  
Multidrug Resistant ◽  
Resistant Bacteria ◽  
Therapeutic Approaches ◽  
Antibiotic Resistant ◽  
Content Type ◽  
Carbapenem Resistant ◽  
Surface Polysaccharides ◽  
Resistant Strains ◽  
Murine Monoclonal Antibodies

ABSTRACT Antibiotics and vaccines have greatly impacted human health in the last century by dramatically reducing the morbidity and mortality associated with infectious diseases. The recent challenge posed by the emergence of multidrug-resistant bacteria could possibly be addressed by novel immune prophylactic and therapeutic approaches. Among the newly threatening pathogens, Klebsiella pneumoniae is particularly worrisome in the nosocomial setting, and its surface polysaccharides are regarded as promising antigen candidates. The majority of Klebsiella carbapenem-resistant strains belong to the sequence type 158 (ST258) lineage, with two main clades expressing capsular polysaccharides CPS1 and CPS2. In a recent article, S. D. Kobayashi and colleagues (mBio 9:e00297-18, 2018, https://doi.org/10.1128/mBio.00297-18) show that CPS2-specific IgGs render ST258 clade 2 bacteria more sensitive to human serum and phagocytic killing. E. Diago-Navarro et al. (mBio 9:e00091-18, 2018, https://doi.org/10.1128/mBio.00091-18) generated two murine monoclonal antibodies recognizing distinct glycotopes of CPS2 that presented functional activity against multiple ST258 strains. These complementary studies represent a step toward the control of this dangerous pathogen.

scholarly journals Impact of Baseline Characteristics on Future Episodes of Bloodstream Infections: Multistate Model in Septic Patients With Bloodstream Infections

2019 ◽  
Author(s):  
M Cristina Vazquez Guillamet ◽  
Rodrigo Vazquez ◽  
Jonas Noe ◽  
Scott T Micek ◽  
Victoria J Fraser ◽  
...  
Keyword(s):  
Risk Factors ◽  
Blood Culture ◽  
Bloodstream Infections ◽  
Baseline Risk ◽  
Resistant Bacteria ◽  
Beta Lactam ◽  
Candida Spp ◽  
Patients At Risk ◽  
Baseline Characteristics ◽  
Inappropriate Antibiotics

Abstract Background Looking only at the index infection, studies have described risk factors for infections caused by resistant bacteria. We hypothesized that septic patients with bloodstream infections may transition across states characterized by different microbiology and that their trajectory is not uniform. We also hypothesized that baseline risk factors may influence subsequent blood culture results. Methods All adult septic patients with positive blood cultures over a 7-year period were included in the study. Baseline risk factors were recorded. We followed all survivors longitudinally and recorded subsequent blood culture results. We separated states into bacteremia caused by gram-positive cocci, susceptible gram-negative bacilli (sGNB), resistant GNB (rGNB), and Candida spp. Detrimental transitions were considered when transitioning to a culture with a higher mortality risk (rGNB and Candida spp.). A multistate Markov-like model was used to determine risk factors associated with detrimental transitions. Results A total of 990 patients survived and experienced at least 1 transition, with a total of 4282 transitions. Inappropriate antibiotics, previous antibiotic exposure, and index bloodstream infection caused by either rGNB or Candida spp. were associated with detrimental transitions. Double antibiotic therapy (beta-lactam plus either an aminoglycoside or a fluoroquinolone) protected against detrimental transitions. Conclusion Baseline characteristics that include prescribed antibiotics can identify patients at risk for subsequent bloodstream infections caused by resistant bacteria. By altering the initial treatment, we could potentially influence future bacteremic states.

scholarly journals Forceps, Actual Use, and Potential Cesarean Section Prevention: Study in a Selected Mexican Population

2015 ◽  
Vol 2015 ◽  
pp. 1-5 ◽  
Author(s):  
Rodrigo Ayala-Yáñez ◽  
Paulette Bayona-Soriano ◽  
Arturo Hernández-Jimenez ◽  
Alejandra Contreras-Rendón ◽  
Paulina Chabat-Manzanera ◽  
...  
Keyword(s):  
Cesarean Section ◽  
Medical Center ◽  
Prevention Study ◽  
Perineal Tear ◽  
Safe Alternative ◽  
Fourth Degree ◽  
Actual Use ◽  
Life Threatening ◽  
Prophylactic Use ◽  
Threatening Situation

Objective. Assessment of the frequency of complications observed with various forceps and operative vaginal delivery (OVD) techniques performed at the ABC Medical Center (Mexico City) to evaluate their safety, bearing in mind the importance of decreasing our country’s high cesarean section incidence.Methods. We reviewed 5,375 deliveries performed between the years 2007 and 2012, only 146 were delivered by OVD.  Results. Only 1.0% of the cases had a serious, life-threatening situation (uterine rupture). The Simpson forceps was the most favored instrument (46%) due to its simplicity of use, effectiveness, and familiarity. Prophylactic use was the most common indication (30.8%) and significant complications observed were vaginal lacerations (p=0.016), relative risk (RR) of 3.4 (95% confidence interval [CI]: 1.15–10.04), and fourth degree perineal tear (p=0.016), RR of 3.4 (95% CI: 1.15–10.04).Conclusions. Forceps use and other OVD techniques are a safe alternative to be considered, diminishing C-section incidence and its complications.

scholarly journals Plasmid Dynamics in KPC-Positive Klebsiella pneumoniae during Long-Term Patient Colonization

mBio ◽  
2016 ◽  
Vol 7 (3) ◽  
Author(s):  
Sean Conlan ◽  
Morgan Park ◽  
Clayton Deming ◽  
Pamela J. Thomas ◽  
Alice C. Young ◽  
...  
Keyword(s):  
Health Care ◽  
Antibiotic Resistance ◽  
Klebsiella Pneumoniae ◽  
Health Care Facilities ◽  
Resistant Bacteria ◽  
Whole Genome ◽  
Antibiotic Resistant ◽  
Carbapenem Resistant ◽  
Clinical Center

ABSTRACT Carbapenem-resistant Klebsiella pneumoniae strains are formidable hospital pathogens that pose a serious threat to patients around the globe due to a rising incidence in health care facilities, high mortality rates associated with infection, and potential to spread antibiotic resistance to other bacterial species, such as Escherichia coli . Over 6 months in 2011, 17 patients at the National Institutes of Health (NIH) Clinical Center became colonized with a highly virulent, transmissible carbapenem-resistant strain of K. pneumoniae . Our real-time genomic sequencing tracked patient-to-patient routes of transmission and informed epidemiologists’ actions to monitor and control this outbreak. Two of these patients remained colonized with carbapenemase-producing organisms for at least 2 to 4 years, providing the opportunity to undertake a focused genomic study of long-term colonization with antibiotic-resistant bacteria. Whole-genome sequencing studies shed light on the underlying complex microbial colonization, including mixed or evolving bacterial populations and gain or loss of plasmids. Isolates from NIH patient 15 showed complex plasmid rearrangements, leaving the chromosome and the bla KPC -carrying plasmid intact but rearranging the two other plasmids of this outbreak strain. NIH patient 16 has shown continuous colonization with bla KPC -positive organisms across multiple time points spanning 2011 to 2015. Genomic studies defined a complex pattern of succession and plasmid transmission across two different K. pneumoniae sequence types and an E. coli isolate. These findings demonstrate the utility of genomic methods for understanding strain succession, genome plasticity, and long-term carriage of antibiotic-resistant organisms. IMPORTANCE In 2011, the NIH Clinical Center had a nosocomial outbreak involving 19 patients who became colonized or infected with bla KPC -positive Klebsiella pneumoniae . Patients who have intestinal colonization with bla KPC -positive K. pneumoniae are at risk for developing infections that are difficult or nearly impossible to treat with existing antibiotic options. Two of those patients remained colonized with bla KPC -positive Klebsiella pneumoniae for over a year, leading to the initiation of a detailed genomic analysis exploring mixed colonization, plasmid recombination, and plasmid diversification. Whole-genome sequence analysis identified a variety of changes, both subtle and large, in the bla KPC -positive organisms. Long-term colonization of patients with bla KPC -positive Klebsiella pneumoniae creates new opportunities for horizontal gene transfer of plasmids encoding antibiotic resistance genes and poses complications for the delivery of health care.

scholarly journals Diagnosis of Bloodstream Infections in Children

2016 ◽  
Vol 54 (6) ◽  
pp. 1418-1424 ◽  
Author(s):  
Jennifer Dien Bard ◽  
Erin McElvania TeKippe
Keyword(s):  
Blood Culture ◽  
Clinical Microbiology ◽  
Bloodstream Infections ◽  
Clinical Microbiology Laboratory ◽  
Microbiology Laboratory ◽  
Diagnostic Approaches ◽  
Pediatric Populations ◽  
Culture Specimen

Identification of bloodstream infections is among the most critical tasks performed by the clinical microbiology laboratory. While the criteria for achieving an adequate blood culture specimen in adults have been well described, there is much more ambiguity in pediatric populations. This minireview focuses on the available pediatric literature pertaining to the collection of an optimal blood culture specimen, including timing, volume, and bottle selection, as well as rapid diagnostic approaches and their role in the management of pediatric bloodstream infections.

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