scholarly journals Whole-Genome Sequencing of a Large Panel of Contemporary Neisseria gonorrhoeae Clinical Isolates Indicates that a Wild-Type mtrA Gene Is Common: Implications for Inducible Antimicrobial Resistance

2017 ◽  
Vol 61 (4) ◽  
Author(s):  
Eshaw Vidyaprakash ◽  
A. Jeanine Abrams ◽  
William M. Shafer ◽  
David L. Trees
Keyword(s):  
Antimicrobial Resistance ◽  
Neisseria Gonorrhoeae ◽  
Whole Genome Sequencing ◽  
Genome Sequencing ◽  
Clinical Isolates ◽  
Whole Genome ◽  
Wild Type ◽  
Large Panel

scholarly journals Neisseria gonorrhoeae clustering to reveal major European whole-genome-sequencing-based genogroups in association with antimicrobial resistance

2020 ◽  
Author(s):  
Miguel Pinto ◽  
Vítor Borges ◽  
Joana Isidro ◽  
João Carlos Rodrigues ◽  
Luís Vieira ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Neisseria Gonorrhoeae ◽  
Whole Genome Sequencing ◽  
Genome Sequencing ◽  
Type Species ◽  
Transmitted Disease ◽  
Epidemiological Surveillance ◽  
Whole Genome ◽  
Content Type ◽  
Link Type

Neisseria gonorrhoeae , the bacterium responsible for the sexually transmitted disease gonorrhoea, has shown an extraordinary ability to develop antimicrobial resistance (AMR) to multiple classes of antimicrobials. With no available vaccine, managing N. gonorrhoeae infections demands effective preventive measures, antibiotic treatment and epidemiological surveillance. The latter two are progressively being supported by the generation of whole-genome sequencing (WGS) data on behalf of national and international surveillance programmes. In this context, this study aims to perform N. gonorrhoeae clustering into genogroups based on WGS data, for enhanced prospective laboratory surveillance. Particularly, it aims to identify the major circulating WGS-genogroups in Europe and to establish a relationship between these and AMR. Ultimately, it enriches public databases by contributing with WGS data from Portuguese isolates spanning 15 years of surveillance. A total of 3791 carefully inspected N. gonorrhoeae genomes from isolates collected across Europe were analysed using a gene-by-gene approach (i.e. using cgMLST). Analysis of cluster composition and stability allowed the classification of isolates into a two-step hierarchical genogroup level determined by two allelic distance thresholds revealing cluster stability. Genogroup clustering in general agreed with available N. gonorrhoeae typing methods [i.e. MLST (multilocus sequence typing), NG-MAST ( N. gonorrhoeae multi-antigen sequence typing) and PubMLST core-genome groups], highlighting the predominant genogroups circulating in Europe, and revealed that the vast majority of the genogroups present a dominant AMR profile. Additionally, a non-static gene-by-gene approach combined with a more discriminatory threshold for potential epidemiological linkage enabled us to match data with previous reports on outbreaks or transmission chains. In conclusion, this genogroup assignment allows a comprehensive analysis of N. gonorrhoeae genetic diversity and the identification of the WGS-based genogroups circulating in Europe, while facilitating the assessment (and continuous monitoring) of their frequency, geographical dispersion and potential association with specific AMR signatures. This strategy may benefit public-health actions through the prioritization of genogroups to be controlled, the identification of emerging resistance carriage, and the potential facilitation of data sharing and communication.

scholarly journals Burkholderia pseudomallei Isolates from Sarawak, Malaysian Borneo, Are Predominantly Susceptible to Aminoglycosides and Macrolides

2013 ◽  
Vol 58 (1) ◽  
pp. 162-166 ◽  
Author(s):  
Yuwana Podin ◽  
Derek S. Sarovich ◽  
Erin P. Price ◽  
Mirjam Kaestli ◽  
Mark Mayo ◽  
...  
Keyword(s):  
Whole Genome Sequencing ◽  
Genome Sequencing ◽  
Burkholderia Pseudomallei ◽  
Sequence Type ◽  
Clinical Isolates ◽  
Whole Genome ◽  
Wild Type ◽  
Content Type ◽  
District Hospitals ◽  
Malaysian Borneo

ABSTRACTMelioidosis is a potentially fatal disease caused by the saprophytic bacteriumBurkholderia pseudomallei. Resistance to gentamicin is generally a hallmark ofB. pseudomallei, and gentamicin is a selective agent in media used for diagnosis of melioidosis. In this study, we determined the prevalence and mechanism of gentamicin susceptibility found inB. pseudomalleiisolates from Sarawak, Malaysian Borneo. We performed multilocus sequence typing and antibiotic susceptibility testing on 44B. pseudomalleiclinical isolates from melioidosis patients in Sarawak district hospitals. Whole-genome sequencing was used to identify the mechanism of gentamicin susceptibility. A novel allelic-specific PCR was designed to differentiate gentamicin-sensitive isolates from wild-typeB. pseudomallei. A reversion assay was performed to confirm the involvement of this mechanism in gentamicin susceptibility. A substantial proportion (86%) ofB. pseudomalleiclinical isolates in Sarawak, Malaysian Borneo, were found to be susceptible to the aminoglycoside gentamicin, a rare occurrence in other regions whereB. pseudomalleiis endemic. Gentamicin sensitivity was restricted to genetically related strains belonging to sequence type 881 or its single-locus variant, sequence type 997. Whole-genome sequencing identified a novel nonsynonymous mutation withinamrB, encoding an essential component of the AmrAB-OprA multidrug efflux pump. We confirmed the role of this mutation in conferring aminoglycoside and macrolide sensitivity by reversion of this mutation to the wild-type sequence. Our study demonstrates that alternativeB. pseudomalleiselective media without gentamicin are needed for accurate melioidosis laboratory diagnosis in Sarawak. This finding may also have implications for environmental sampling of other locations to test forB. pseudomalleiendemicity.

scholarly journals Comparison of Antimicrobial Resistance and Pan-Genome of Clinical and Non-Clinical Enterococcus cecorum from Poultry Using Whole-Genome Sequencing

Foods ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 686
Author(s):  
Poonam Sharma ◽  
Sushim K. Gupta ◽  
John B. Barrett ◽  
Lari M. Hiott ◽  
Tiffanie A. Woodley ◽  
...  
Keyword(s):  
Antimicrobial Resistance ◽  
Whole Genome Sequencing ◽  
Genome Sequencing ◽  
Resistance Genes ◽  
Gene Families ◽  
Clinical Isolates ◽  
Whole Genome ◽  
Pan Genome ◽  
Antimicrobial Resistance Genes ◽  
Enterococcus Cecorum

Enterococcus cecorum is an emerging avian pathogen, particularly in chickens, but can be found in both diseased (clinical) and healthy (non-clinical) poultry. To better define differences between E. cecorum from the two groups, whole-genome sequencing (WGS) was used to identify and compare antimicrobial resistance genes as well as the pan-genome among the isolates. Eighteen strains selected from our previous study were subjected to WGS using Illumina MiSeq and comparatively analyzed. Assembled contigs were analyzed for resistance genes using ARG-ANNOT. Resistance to erythromycin was mediated by ermB, ermG, and mefA, in clinical isolates and ermB and mefA, in non-clinical isolates. Lincomycin resistance genes were identified as linB, lnuB, lnuC, and lnuD with lnuD found only in non-clinical E. cecorum; however, lnuB and linB were found in only one clinical isolate. For both groups of isolates, kanamycin resistance was mediated by aph3-III, while tetracycline resistance was conferred by tetM, tetO, and tetL. No mutations or known resistance genes were found for isolates resistant to either linezolid or chloramphenicol, suggesting possible new mechanisms of resistance to these drugs. A comparison of WGS results confirmed that non-clinical isolates contained more resistance genes than clinical isolates. The pan-genome of clinical and non-clinical isolates resulted in 3651 and 4950 gene families, respectively, whereas the core gene sets were comprised of 1559 and 1534 gene families in clinical and non-clinical isolates, respectively. Unique genes were found more frequently in non-clinical isolates than clinical. Phylogenetic analysis of the isolates and all the available complete and draft genomes showed no correlation between healthy and diseased poultry. Additional genomic comparison is required to elucidate genetic factors in E. cecorum that contribute to disease in poultry.

scholarly journals 1436. Use of Whole Genome Sequencing to Characterize Antimicrobial-resistant Salmonella Berta Isolates from Clinical and Retail Meat Sources

2020 ◽  
Vol 7 (Supplement_1) ◽  
pp. S722-S723
Author(s):  
Nkuchia M M’ikanatha ◽  
Rachael Jacques ◽  
David Faucette ◽  
Dettinger Lisa ◽  
Kevin Libuit ◽  
...  
Keyword(s):  
Single Nucleotide Polymorphism ◽  
Antimicrobial Resistance ◽  
Whole Genome Sequencing ◽  
Genome Sequencing ◽  
Clinical Isolates ◽  
Health Concern ◽  
Whole Genome ◽  
Nucleotide Polymorphism ◽  
Single Nucleotide ◽  
Retail Meat

Abstract Background Antimicrobial resistance (AMR) in foodborne pathogens of animal origin, including non-typhoidal Salmonella (NTS) are a public health concern. Pennsylvania conducts integrated surveillance for AMR in NTS isolates from human and animal sources in collaboration with the National Antimicrobial Resistance Monitoring System (NARMS). Methods During 2009-2014, Salmonella enterica isolates from various types of meat purchased from randomly selected retail outlets in southeastern Pennsylvania were analyzed by pulsed-field gel electrophoresis (PFGE). We compared PFGE patterns from meat with clinical isolates in the Pennsylvania surveillance database. All meat isolates and a subset of matched clinical isolates were tested for susceptibility to antimicrobial agents. Eleven isolates with indistinguishable PFGE patterns were analyzed by whole genome sequencing (WGS). Sequence data were uploaded to the FDA’s GalaxyTrakr platform for quality assessment, genome assembly, AMR gene detection, and phylogenetic inference via single-nucleotide polymorphism (SNP) analysis. Results PFGE patterns of 86 (48.6%) of 177 meat isolates had PFGE matches to 1,665 clinical isolates; 40 distinct PFGE patterns were represented among the shared patterns. Seventeen (43%) of the 40 shared PFGE patterns (with ≥1 isolate(s) from both sources) were considered multi-drug resistant (MDR). Among the 48 S. Berta pattern JAXX01.0001 isolates, 5 (10.9%) and 2 (100%) from human and meat sources respectively were MDR including resistance to amoxicillin and ceftriaxone. WGS analysis of one isolate from ground turkey meat (PNUSAS061602) was genetically related to clinical isolates including two within 9 and 11 SNPs [Figure]. Presence of genes that hydrolyze extended spectrum cephalosporins (ESC), [blaCMY, blaHERA, or blaTEM], was identified in eight (two meat and six clinical) isolates. One meat isolate was resistant to six antibiotics including ceftriaxone. Figure 2. Single nucleotide polymorphism (SNP) distance matrix showing relatedness in non-typhoidal Salmonella isolates from retail meat (n=2) and human (n=9) sources — Pennsylvania, 2010-2014. One S Berta from retail meat was separated from two clinical two clinical isolates by 9 and 11 SNPs. Second isolate from meat was separated from those associated with human infections by 14 (n=1), 17 (n=1) and ≥20 (n=7). Conclusion WGS analysis revealed clinically relevant ESCs genes in closely related S. Berta isolates from human and animal sources. Presence of these genes in NTS highlights the need for enhanced One-Health surveillance and judicious use of antibiotics in humans and food-animal production. Disclosures All Authors: No reported disclosures

scholarly journals Antimicrobial resistance profiling and phylogenetic analysis of Neisseria gonorrhoeae clinical isolates from Kenya in a resource-limited setting

2020 ◽  
Author(s):  
Meshack O Juma ◽  
Arun Sankaradoss ◽  
Redcliff Ndombi ◽  
Patrick Mwaura ◽  
Tina Damodar ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Phylogenetic Analysis ◽  
Antimicrobial Resistance ◽  
Neisseria Gonorrhoeae ◽  
Whole Genome Sequencing ◽  
Genome Sequencing ◽  
Whole Genome ◽  
Resource Limited Settings ◽  
Resource Limited ◽  
The World

Background Africa has one of the highest incidences of gonorrhoea, but not much information is available on the relatedness with strains from other geographical locations. Antimicrobial resistance (AMR) in Neisseria gonorrhoeae is a major public health threat, with the bacteria gaining resistance to most of the available antibiotics, compromising treatment across the world. Whole-genome sequencing is an efficient way of predicting AMR determinants and their spread in the human population. Previous studies on Kenyan gonococcal samples have focused on plasmid-mediated drug resistance and fluoroquinolone resistance using Illumina sequencing. Recent advances in next-generation sequencing technologies like Oxford Nanopore Technology (ONT) have helped in the generation of longer reads of DNA in a shorter duration with lower cost. However, long-reads are error-prone. The increasing accuracy of base-calling algorithms, high throughput, error-correction strategies, and ease of using the mobile sequencer in remote areas is leading to the adoption of the MinION sequencer (ONT), for routine microbial genome sequencing. Methods To investigate whether MinION-only sequencing is sufficient for diagnosis, genome sequencing and downstream analysis like inferring phylogenetic relationships and detection of AMR in resource-limited settings, we sequenced the genomes of fourteen clinical isolates suspected to be N. gonorrhoeae from Nairobi, Kenya. The isolates were tested using standard bacteriological methods for identification, interpretted using analytical profile index and antibiotic susceptibility tests had indicated ciprofloxacin and gentamycin resistance. Using whole genome sequencing, the isolates were confirmed to be cases of N. gonorrhoeae (n=12), Additionally, we identified reads from N. meningitidis (n=2) and both of N. gonorrhoeae and Moraxella osloensis (n=3) in the sample (co-infections) respectively, which have been implicated in sexually transmitted infections in the recent years. The near-complete N. gonorrhoeae genomes (n=10) were anaysed further for mutations/factors causing AMR using an in-house database of mutations curated from the literature. We attempted to understand the basis of drug resistance using homology modelling of AMR proteins, using known structures from other bacteria. Results We observe that Ciprofloxacin resistance is associated with multiple mutations in both gyrA and parC. We identified mutations conferring tetracycline (rpsJ) and Sulfonamide (folA) resistance in all the isolates and plasmids encoding beta-lactamase and tet(M) were identified in almost all of the strains. Phylogenetic analysis clustered the nine isolates into clades containing previously sequenced genomes from Kenya and countries across the world. Conclusion Here, we demonstrate the utility of mobile DNA sequencing technology supplemented with reference-based assembly in sequence typing and elucidating the basis of AMR. Bioinformatics profiling to predict AMR can be used along with routine AMR susceptibily tests in clinics. The workflow followed in the study, including AMR mutation dataset creation and the genome identification, assembly and analysis, can be used for the genome assembly and analysis of any clinical isolate. Further studies are required to determine the utility of real-time sequencing in the outbreak investigations, diagnosis and management of infections, especially in resource-limited settings.

scholarly journals Expanding U.S. Laboratory Capacity for Neisseria gonorrhoeae Antimicrobial Susceptibility Testing and Whole-Genome Sequencing through the CDC's Antibiotic Resistance Laboratory Network

2020 ◽  
Vol 58 (4) ◽  
Author(s):  
Ellen N. Kersh ◽  
Cau D. Pham ◽  
John R. Papp ◽  
Robert Myers ◽  
Richard Steece ◽  
...  
Keyword(s):  
Public Health ◽  
Antibiotic Resistance ◽  
Neisseria Gonorrhoeae ◽  
Whole Genome Sequencing ◽  
Genome Sequencing ◽  
Antimicrobial Susceptibility ◽  
Susceptibility Testing ◽  
Whole Genome ◽  
Content Type ◽  
Laboratory Capacity

ABSTRACT U.S. gonorrhea rates are rising, and antibiotic-resistant Neisseria gonorrhoeae (AR-Ng) is an urgent public health threat. Since implementation of nucleic acid amplification tests for N. gonorrhoeae identification, the capacity for culturing N. gonorrhoeae in the United States has declined, along with the ability to perform culture-based antimicrobial susceptibility testing (AST). Yet AST is critical for detecting and monitoring AR-Ng. In 2016, the CDC established the Antibiotic Resistance Laboratory Network (AR Lab Network) to shore up the national capacity for detecting several resistance threats including N. gonorrhoeae. AR-Ng testing, a subactivity of the CDC’s AR Lab Network, is performed in a tiered network of approximately 35 local laboratories, four regional laboratories (state public health laboratories in Maryland, Tennessee, Texas, and Washington), and the CDC’s national reference laboratory. Local laboratories receive specimens from approximately 60 clinics associated with the Gonococcal Isolate Surveillance Project (GISP), enhanced GISP (eGISP), and the program Strengthening the U.S. Response to Resistant Gonorrhea (SURRG). They isolate and ship up to 20,000 isolates to regional laboratories for culture-based agar dilution AST with seven antibiotics and for whole-genome sequencing of up to 5,000 isolates. The CDC further examines concerning isolates and monitors genetic AR markers. During 2017 and 2018, the network tested 8,214 and 8,628 N. gonorrhoeae isolates, respectively, and the CDC received 531 and 646 concerning isolates and 605 and 3,159 sequences, respectively. In summary, the AR Lab Network supported the laboratory capacity for N. gonorrhoeae AST and associated genetic marker detection, expanding preexisting notification and analysis systems for resistance detection. Continued, robust AST and genomic capacity can help inform national public health monitoring and intervention.

Sign in / Sign up

Export Citation Format

Share Document