Prevalence and molecular epidemiology of mcr-1-positive Klebsiella pneumoniae in healthy adults from China

2020 ◽  
Vol 75 (9) ◽  
pp. 2485-2494 ◽  
Author(s):  
Jiayue Lu ◽  
Ning Dong ◽  
Congcong Liu ◽  
Yu Zeng ◽  
Qiaoling Sun ◽  
...  
Keyword(s):  
Klebsiella Pneumoniae ◽  
Antimicrobial Susceptibility ◽  
Susceptibility Testing ◽  
Mainland China ◽  
Healthy Adults ◽  
Antimicrobial Susceptibility Testing ◽  
Pcr Analysis ◽  
Resistant Bacteria ◽  
Healthy Individuals ◽  
Low Prevalence

Abstract Objectives To investigate the nationwide prevalence of mcr-1-positive Klebsiella pneumoniae (MCRPKP) strains among healthy adults in China and identify their phenotypic and genomic characterizations. Methods A total of 7401 rectal swab samples were collected from healthy individuals in 30 hospitals located in 30 provinces and municipalities of mainland China in 2016. Colistin-resistant bacteria were enriched in colistin-supplemented lysogeny broth. MCRPKP strains were isolated and characterized with MALDI-TOF MS, PCR analysis and antimicrobial susceptibility testing. The genomic characteristics of MCRPKP strains were determined by WGS and bioinformatics analysis. Results Seven MCRPKP strains and one mcr-1-positive Klebsiella variicola strain were selectively isolated from six locales (three from Henan and one from each of Tianjin, Jiangxi, Yunnan, Gansu and Tibet). Antimicrobial susceptibility testing results indicated that all mcr-1-positive strains were susceptible to meropenem, aztreonam and ceftazidime/avibactam. WGS analysis suggested these strains belonged to seven distinct STs: ST15, ST1425, ST1462, ST273, ST307, ST391 and ST37-SLV. mcr-1 genes were carried by diverse plasmids, including IncHI2 (n = 3), IncX4 (n = 2), IncHI2/IncN (n = 1), IncFIB (n = 1) and one other plasmid type. Two ST15 strains harboured both mcr-1 and mcr-8 genes, which has not been reported before. Conclusions Our data indicated a low prevalence of mcr-1-positive Klebsiella strains (0.11%, 8/7401) in healthy individuals in mainland China and most of these strains remained susceptible to clinically important antibiotics. The prevalence and coexistence of mcr-1 and mcr-8 in K. pneumoniae may further threaten public health through either the food chain or environmental routes.

scholarly journals Emergence and Genomic Characterization of a KPC-2-, NDM-1-, and IMP-4-Producing Klebsiella michiganensis Isolate

2022 ◽  
Vol 12 ◽  
Author(s):  
Yanyan Zhang ◽  
Danxia Gu ◽  
Xuemei Yang ◽  
Yuchen Wu ◽  
Congcong Liu ◽  
...  
Keyword(s):  
Whole Genome Sequencing ◽  
Genome Sequencing ◽  
Antimicrobial Susceptibility ◽  
Colloidal Gold ◽  
Susceptibility Testing ◽  
Antimicrobial Susceptibility Testing ◽  
Pcr Analysis ◽  
Resistant Bacteria ◽  
Whole Genome ◽  
Sequencing Analysis

A rectal swab sample was collected from a patient with Guillain–Barré syndrome and enriched in lysogeny broth. Carbapenem-resistant bacteria were selected by China Blue agar plates containing 0.3 μg/ml meropenem. Carbapenemase-producing Klebsiella michiganensis was identified and characterized by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF), immune colloidal gold technique, a conjugation experiment, PCR analysis, and antimicrobial susceptibility testing. The genome of K. michiganensis was determined by whole genome sequencing. Antimicrobial susceptibility testing showed that the K. michiganensis was resistant to imipenem, meropenem, ertapenem, cefmetazole, ceftazidime, cefotaxime, piperacillin/tazobactam, sulbactam/cefoperazone, ceftazidime/avibactam, cefepime, and aztreonam while susceptible to polymyxin B, ciprofloxacin, tigecycline, and amikacin. Immune colloidal gold technique suggested that this strain co-produced three different carbapenemases [Klebsiella pneumoniae carbapenemase (KPC), New Delhi metallo-beta-lactamase (NDM), and Imipenem (IMP)]. Whole genome sequencing analysis indicated that this strain belonged to ST91, and blaKPC–2, blaNDM–1, and blaIMP–4 were carried on different conjugative plasmids. Besides, the co-existence and transferability of blaKPC–2, blaNDM–1, and blaIMP–4 in K. michiganensis facilitates the potential horizontal dissemination and nosocomial spread of resistance genes among multidrug-resistant organisms.

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 Antibiotic Resistance among Escherichia coli Isolates from Hospital Wastewater Compared to Community Wastewater

Water ◽  
2021 ◽  
Vol 13 (23) ◽  
pp. 3449
Author(s):  
Cristina-Mirabela Gaşpar ◽  
Ludovic Toma Cziszter ◽  
Cristian Florin Lăzărescu ◽  
Ioan Ţibru ◽  
Marius Pentea ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Antimicrobial Susceptibility ◽  
Susceptibility Testing ◽  
Antimicrobial Susceptibility Testing ◽  
Diffusion Method ◽  
Resistant Bacteria ◽  
Hospital Wastewater ◽  
Extended Spectrum Beta Lactamase ◽  
E Coli

This study aimed to compare the antibiotic resistance levels of the indicator bacteria Escherichia coli in wastewater samples collected from two hospitals and two urban communities. Antimicrobial susceptibility testing was performed on 81 E. coli isolates (47 from hospitals and 34 from communities) using the disc diffusion method according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST) methodology. Ten antibiotics from nine different classes were chosen. The strains isolated from the community wastewater, compared to those from the hospital wastewater, were not resistant to gentamicin (p = 0.03), but they showed a significantly higher susceptibility—increased exposure to ceftazidime (p = 0.001). Multidrug resistance was observed in 85.11% of the hospital wastewater isolates and 73.53% of the community isolates (p > 0.05). The frequency of the presumed carbapenemase-producing E. coli was higher among the community isolates (76.47% compared to 68.09%) (p > 0.05), whereas the frequency of the presumed extended-spectrum beta-lactamase (ESBL)-producing E. coli was higher among the hospital isolates (21.28% compared to 5.88%) (p > 0.05). The antibiotic resistance rates were high in both the hospital and community wastewaters, with very few significant differences between them, so the community outlet might be a source of resistant bacteria that is at least as important as the well-recognised hospitals.

scholarly journals Antimikrobiyal Aktivitenin Belirlenmesinde Cross-Streak Metodu Kullanımı

2019 ◽  
Vol 7 (sp1) ◽  
pp. 160
Author(s):  
Mustafa Ersal
Keyword(s):  
Antimicrobial Susceptibility ◽  
Antimicrobial Agents ◽  
Susceptibility Testing ◽  
Clinical Laboratory ◽  
Antimicrobial Susceptibility Testing ◽  
Rapid Screening ◽  
Resistant Bacteria ◽  
Production Environment ◽  
Specific Test ◽  
The Cross

Antimicrobial susceptibility testing can be used for prediction of therapeutic results, epidemiology and drug discovery. Microbial infections are an important problem which have developed resistance towards antimicrobial agents. Otherwise, efficacy of these agents is considerable with treatment failures associated with multidrug-resistant bacteria and it has become a global concern to public health. Therefore, explore the new antimicrobial agents and widely use of antimicrobial susceptibility need to be developed. There are many techniques for the determination of antimicrobial activity. Many of these techniques, which are applied to inhibit sensitive microorganisms, are based on diffusion-related methods in the solid or semi-solid production environment. Cross-streak among these techniques is an easy technique that allows for relatively rapid screening of cultures in research for the discovery of the new antibiotics. However, the biggest disadvantage of the Cross-streak test is the difficulty in obtaining quantitative data. Because the edges of the inhibition zone are usually very fuzzy and unclear. Some antimicrobial susceptibility testing techniques were standardized by Clinical Laboratory Standards Institute (CLSI) and European Committee on Antimicrobial Susceptibility Testing (EUCAST) to determine the striking steps in this area. This testing procedure requires the use of specific test conditions and methods. In addition, the medium, incubation conditions and time are among these requirements. It is important to understand and develop the Cross-streak method from the currently used activity determination methods.

scholarly journals A Rapid Single-Cell Antimicrobial Susceptibility Testing Workflow for Bloodstream Infections

Biosensors ◽  
2021 ◽  
Vol 11 (8) ◽  
pp. 288
Author(s):  
Britney Forsyth ◽  
Peter Torab ◽  
Jyong-Huei Lee ◽  
Tyler Malcom ◽  
Tza-Huei Wang ◽  
...  
Keyword(s):  
Single Cell ◽  
Antimicrobial Susceptibility ◽  
Susceptibility Testing ◽  
Rapid Determination ◽  
Bloodstream Infections ◽  
Antimicrobial Susceptibility Testing ◽  
Resistant Bacteria ◽  
Bloodborne Pathogens ◽  
The Matrix ◽  
Single Cell Trapping

Bloodstream infections are a significant cause of morbidity and mortality worldwide. The rapid initiation of effective antibiotic treatment is critical for patients with bloodstream infections. However, the diagnosis of bloodborne pathogens is largely complicated by the matrix effect of blood and the lengthy blood tube culture procedure. Here we report a culture-free workflow for the rapid isolation and enrichment of bacterial pathogens from whole blood for single-cell antimicrobial susceptibility testing (AST). A dextran sedimentation step reduces the concentration of blood cells by 4 orders of magnitude in 20–30 min while maintaining the effective concentration of bacteria in the sample. Red blood cell depletion facilitates the downstream centrifugation-based enrichment step at a sepsis-relevant bacteria concentration. The workflow is compatible with common antibiotic-resistant bacteria and does not influence the minimum inhibitory concentrations. By applying a microfluidic single-cell trapping device, we demonstrate the workflow for the rapid determination of bacterial infection and antimicrobial susceptibility testing at the single-cell level. The entire workflow from blood to categorical AST result can be completed in less than two hours.

scholarly journals Rapid antimicrobial susceptibility testing on positive blood cultures through an innovative light scattering technology: performances and turnaround time evaluation

2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Lidvine Boland ◽  
Corentin Streel ◽  
Hélène De Wolf ◽  
Hector Rodriguez ◽  
Alexia Verroken
Keyword(s):  
Light Scattering ◽  
Antimicrobial Susceptibility ◽  
Susceptibility Testing ◽  
Turnaround Time ◽  
Blood Cultures ◽  
Antimicrobial Susceptibility Testing ◽  
Resistant Bacteria ◽  
Gram Positive ◽  
Gram Negative ◽  
Gram Positive Cocci

Abstract Background A bacteremia diagnosis with speeded-up identification and antimicrobial susceptibility testing (AST) is mandatory to adjust empirical broad-spectrum antibiotherapy and avoid the emergence of multi-resistant bacteria. Alfred 60AST (Alifax, Polverara, PD, Italy) is an innovative automated system based on light scattering measurements allowing direct AST from positive blood cultures with rapid results. In this study we aimed to evaluate the system’s performances and turnaround time (TAT) compared to routine AST. Methods The study was conducted during 2 non-consecutive 3-month periods at the microbiology laboratory of the Cliniques universitaires Saint-Luc. All blood cultures detected positive in the 0 AM–10 AM time frame with a pure Gram-positive cocci or Gram-negative bacilli stain were included for Alfred 60AST testing. Two customized EUCAST antibiotic panels were set up composed of 1) a “Gram-negative” panel including cefuroxime, ceftazidime Enterobacteriaceae, piperacillin-tazobactam Enterobacteriaceae, ciprofloxacine, and ceftazidime Pseudomonas 2) a “Gram-positive” panel including cefoxitin Staphylococcus aureus, cefoxitin coagulase-negative (CNS) Staphylococci and ampicillin Enterococci. Categorical agreement (CA), very major errors (VME), major errors (ME), minor errors (mE) and TAT to Alfred 60AST results were calculated in comparison with AST results obtained from direct testing on positive blood cultures with the Phoenix system (Becton Dickinson, Franklin Lakes, NJ, USA). Results Five hundred seventy and one hundred nine antibiotics were evaluated on respectively 166 Gram-negative bacilli and 109 Gram-positive cocci included in the studied population. During the first study period regarding Gram-negative strains a CA of 89.5% was obtained with a high rate of VME (19 and 15.4% respectively) for cefuroxime and piperacillin-tazobactam Enterobacteriaceae. Considering this, Alifax reviewed these antibiotics’ formulations improving Gram-negative bacilli total CA to 92.2% with no VME during the second study period. For Gram-positive cocci, total CA was 88.1% with 2.3% VME, 13.8% ME (mainly cefoxitin CNS) and 12% mE rates both study periods combined. Median TAT to AST results was 5 h with Alfred versus 12 h34 with Phoenix. Conclusion The Alfred 60AST system shows correct yet improvable microbiological performances and a major TAT reduction compared to direct automated AST testing. Clinical studies measuring the impact of the approach on antibiotic management of patients with bacteremia are recommended.

scholarly journals EUCAST rapid antimicrobial susceptibility testing (RAST) in blood cultures: validation in 55 European laboratories

2020 ◽  
Vol 75 (11) ◽  
pp. 3230-3238
Author(s):  
Anna Åkerlund ◽  
Emma Jonasson ◽  
Erika Matuschek ◽  
Lena Serrander ◽  
Martin Sundqvist ◽  
...  
Keyword(s):  
Antimicrobial Susceptibility ◽  
Susceptibility Testing ◽  
Total Error ◽  
Bloodstream Infections ◽  
Error Rates ◽  
Blood Cultures ◽  
Antimicrobial Susceptibility Testing ◽  
Fine Tuning ◽  
Resistant Bacteria ◽  
Disc Diffusion

Abstract Objectives When bloodstream infections are caused by resistant bacteria, rapid antimicrobial susceptibility testing (RAST) is important for adjustment of therapy. The EUCAST RAST method, directly from positive blood cultures, was validated in a multi-laboratory study in Europe. Methods RAST was performed in 40 laboratories in northern Europe (NE) and 15 in southern Europe (SE) from clinical blood cultures positive for Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Staphylococcus aureus or Streptococcus pneumoniae. Categorical results at 4, 6 and 8 h of incubation were compared with results for EUCAST standard 16–20 h disc diffusion. The method, preliminary breakpoints and the performance of the laboratories were evaluated. Results The total number of isolates was 833/318 in NE/SE. The number of zone diameters that could be read (88%, 96% and 99%) and interpreted (70%, 81% and 85%) increased with incubation time (4, 6 and 8 h). The categorical agreement was acceptable, with total error rates in NE/SE of 2.4%/4.9% at 4 h, 1.1%/3.5% at 6 h and 1.1%/3.3% at 8 h. False susceptibility at 4, 6 and 8 h of incubation was below 0.3% and 1.1% in NE and SE, respectively, and the corresponding percentages for false resistance were below 1.9% and 2.8%. After fine-tuning breakpoints, more zones could be interpreted (73%, 89% and 93%), with only marginally affected error rates. Conclusions The EUCAST RAST method can be implemented in routine laboratories without major investments. It provides reliable antimicrobial susceptibility testing results for relevant bloodstream infection pathogens after 4–6 h of incubation.

scholarly journals RNA markers for ultra-rapid molecular antimicrobial susceptibility testing in fluoroquinolone-treated Klebsiella pneumoniae

2020 ◽  
Vol 75 (7) ◽  
pp. 1747-1755 ◽  
Author(s):  
Xi Yang ◽  
Marjan M Hashemi ◽  
Nadya Andini ◽  
Michelle M Li ◽  
Shuzhen Kuang ◽  
...  
Keyword(s):  
Klebsiella Pneumoniae ◽  
Rna Sequencing ◽  
Antimicrobial Susceptibility ◽  
Susceptibility Testing ◽  
Expression Profiles ◽  
Transcript Level ◽  
Bioinformatic Analysis ◽  
Antimicrobial Susceptibility Testing ◽  
Marker Selection ◽  
Appropriate Treatment

Abstract Objectives Traditional antimicrobial susceptibility testing (AST) is growth dependent and time-consuming. With rising rates of drug-resistant infections, a novel diagnostic method is critically needed that can rapidly reveal a pathogen’s antimicrobial susceptibility to guide appropriate treatment. Recently, RNA sequencing has been identified as a powerful diagnostic tool to explore transcriptional gene expression and improve AST. Methods RNA sequencing was used to investigate the potential of RNA markers for rapid molecular AST using Klebsiella pneumoniae and ciprofloxacin as a model. Downstream bioinformatic analysis was applied for optimal marker selection. Further validation on 11 more isolates of K. pneumoniae was performed using quantitative real-time PCR. Results From RNA sequencing, we identified RNA signatures that were induced or suppressed following exposure to ciprofloxacin. Significant shifts at the transcript level were observed as early as 10 min after antibiotic exposure. Lastly, we confirmed marker expression profiles with concordant MIC results from traditional culture-based AST and validated across 11 K. pneumoniae isolates. recA, coaA and metN transcripts harbour the most sensitive susceptibility information and were selected as our top markers. Conclusions Our results suggest that RNA signature is a promising approach to AST development, resulting in faster clinical diagnosis and treatment of infectious disease. This approach is potentially applicable in other models including other pathogens exposed to different classes of antibiotics.

scholarly journals Genomic evolution of the globally disseminated multidrug-resistant Klebsiella pneumoniae clonal group 147

2022 ◽  
Vol 8 (1) ◽  
Author(s):  
Carla Rodrigues ◽  
Siddhi Desai ◽  
Virginie Passet ◽  
Devarshi Gajjar ◽  
Sylvain Brisse
Keyword(s):  
Klebsiella Pneumoniae ◽  
Resistance Genes ◽  
Antimicrobial Susceptibility ◽  
Type Species ◽  
Susceptibility Testing ◽  
Multidrug Resistant ◽  
Antimicrobial Susceptibility Testing ◽  
Content Type ◽  
Genetic Elements ◽  
Link Type

The rapid emergence of multidrug-resistant Klebsiella pneumoniae is being driven largely by the spread of specific clonal groups (CGs). Of these, CG147 includes 7-gene multilocus sequence typing (MLST) sequence types (STs) ST147, ST273 and ST392. CG147 has caused nosocomial outbreaks across the world, but its global population dynamics remain unknown. Here, we report a pandrug-resistant ST147 clinical isolate from India (strain DJ) and define the evolution and global emergence of CG147. Antimicrobial-susceptibility testing following European Committee on Antimicrobial Susceptibility Testing (EUCAST) guidelines and genome sequencing (Illumina and Oxford Nanopore Technologies, Unicycler assembly) were performed on strain DJ. Additionally, we collated 217 publicly available CG147 genomes [National Center for Biotechnology Information (NCBI), May 2019]. CG147 evolution was inferred within a temporal phylogenetic framework (beast) based on a recombination-free sequence alignment (Roary/Gubbins). Comparative genomic analyses focused on resistance and virulence genes and other genetic elements (BIGSdb, Kleborate, PlasmidFinder, phaster, ICEfinder and CRISPRCasFinder). Strain DJ had a pandrug-resistance phenotype. Its genome comprised the chromosome, seven plasmids and one linear phage-plasmid. Four carbapenemase genes were detected: bla NDM-5 and two copies of bla OXA-181 in the chromosome, and a second copy of bla NDM-5 on an 84 kb IncFII plasmid. CG147 genomes carried a mean of 13 acquired resistance genes or mutations; 63 % carried a carbapenemase gene and 83 % harboured bla CTX-M. All CG147 genomes presented GyrA and ParC mutations and a common subtype I-E CRISPR-Cas system. ST392 and ST273 emerged in 2005 and 1995, respectively. ST147, the most represented phylogenetic branch, was itself divided into two main clades with distinct capsular loci: KL64 (74 %, DJ included, emerged in 1994 and disseminated worldwide, with carbapenemases varying among world regions) and KL10 (20 %, emerged in 2002, predominantly found in Asian countries, associated with carbapenemases NDM and OXA-48-like). Furthermore, subclades within ST147-KL64 differed at the yersiniabactin locus, OmpK35/K36 mutations, plasmid replicons and prophages. The absence of IncF plasmids in some subclades was associated with a possible activity of a CRISPR-Cas system. K. pneumoniae CG147 comprises pandrug-resistant or extensively resistant isolates, and carries multiple and diverse resistance genes and mobile genetic elements, including chromosomal bla NDM-5. Its emergence is being driven by the spread of several phylogenetic clades marked by their own genomic features and specific temporo–spatial dynamics. These findings highlight the need for precision surveillance strategies to limit the spread of particularly concerning CG147 subsets.

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