scholarly journals Pilot Evaluation of a Fully Automated Bioinformatics System for Analysis of Methicillin-Resistant Staphylococcus aureus Genomes and Detection of Outbreaks

2019 ◽  
Vol 57 (11) ◽  
Author(s):  
Nicholas M. Brown ◽  
Beth Blane ◽  
Kathy E. Raven ◽  
Narender Kumar ◽  
Danielle Leek ◽  
...  

ABSTRACT Genomic surveillance that combines bacterial sequencing and epidemiological information will become the gold standard for outbreak detection, but its clinical translation is hampered by the lack of automated interpretation tools. We performed a prospective pilot study to evaluate the analysis of methicillin-resistant Staphylococcus aureus (MRSA) genomes using the Next Gen Diagnostics (NGD) automated bioinformatics system. Seventeen unselected MRSA-positive patients were identified in a clinical microbiology laboratory in England over a period of 2 weeks in 2018, and 1 MRSA isolate per case was sequenced on the Illumina MiniSeq instrument. The NGD system automatically activated after sequencing and processed fastq folders to determine species, multilocus sequence type, the presence of a mec gene, antibiotic susceptibility predictions, and genetic relatedness based on mapping to a reference MRSA genome and detection of pairwise core genome single-nucleotide polymorphisms. The NGD system required 90 s per sample to automatically analyze data from each run, the results of which were automatically displayed. The same data were independently analyzed using a research-based approach. There was full concordance between the two analysis methods regarding species (S. aureus), detection of mecA, sequence type assignment, and detection of genetic determinants of resistance. Both analysis methods identified two MRSA clusters based on relatedness, one of which contained 3 cases that were involved in an outbreak linked to a clinic and ward associated with diabetic patient care. We conclude that, in this pilot study, the NGD system provided rapid and accurate data that could support infection control practices.

2014 ◽  
Vol 58 (4) ◽  
pp. 1907-1917 ◽  
Author(s):  
Peter M. Kinnevey ◽  
Anna C. Shore ◽  
Grainne I. Brennan ◽  
Derek J. Sullivan ◽  
Ralf Ehricht ◽  
...  

ABSTRACTClonal replacement of predominant nosocomial methicillin-resistantStaphylococcus aureus(MRSA) strains has occurred several times in Ireland during the last 4 decades. However, little is known about sporadically occurring MRSA in Irish hospitals or in other countries. Eighty-eight representativepvl-negative sporadic MRSA isolates recovered in Irish hospitals between 2000 and 2012 were investigated. These yielded unusual pulsed-field gel electrophoresis and antibiogram-resistogram typing patterns distinct from those of the predominant nosocomial MRSA clone, ST22-MRSA-IV, during the study period. Isolates were characterized byspatyping and DNA microarray profiling for multilocus sequence type (MLST) clonal complex (CC) and/or sequence type (ST) and SCCmectype assignment, as well as for detection of virulence and antimicrobial resistance genes. Conventional PCR-based SCCmecsubtyping was undertaken when necessary. Extensive diversity was detected, including 38spatypes, 13 MLST-CCs (including 18 STs among 62 isolates assigned to STs), and 25 SCCmectypes (including 2 possible novel SCCmecelements and 7 possible novel SCCmecsubtypes). Fifty-four MLST-spa-SCCmectype combinations were identified. Overall, 68.5% of isolates were assigned to nosocomial lineages, with ST8-t190-MRSA-IID/IIE ± SCCM1predominating (17.4%), followed by CC779/ST779-t878-MRSA-ψSCCmec-SCC-SCCCRISPR(7.6%) and CC22/ST22-t032-MRSA-IVh (5.4%). Community-associated clones, including CC1-t127/t386/t2279-MRSA-IV, CC59-t216-MRSA-V, CC8-t008-MRSA-IVa, and CC5-t002/t242-MRSA-IV/V, and putative animal-associated clones, including CC130-t12399-MRSA-XI, ST8-t064-MRSA-IVa, ST398-t011-MRSA-IVa, and CC6-t701-MRSA-V, were also identified. In total, 53.3% and 47.8% of isolates harbored genes for resistance to two or more classes of antimicrobial agents and two or more mobile genetic element-encoded virulence-associated factors, respectively. Effective ongoing surveillance of sporadic nosocomial MRSA is warranted for early detection of emerging clones and reservoirs of virulence, resistance, and SCCmecgenes.


2020 ◽  
Vol 9 (41) ◽  
Author(s):  
Raphael N. Sieber ◽  
Søren Overballe-Petersen ◽  
Hülya Kaya ◽  
Anders R. Larsen ◽  
Andreas Petersen

ABSTRACT Methicillin-resistant Staphylococcus aureus (MRSA) sequence type 630 (ST630) and spa type t4549 is an emerging lineage in Nordic countries, and some representatives carry the CRISPR-Cas system. Here, the complete genome sequences of two isolates from this lineage are presented, comprising chromosomes of 2,918,239 and 2,877,083 nucleotides, respectively, and a 2,473-nucleotide plasmid carrying erm(C).


2017 ◽  
Vol 5 (35) ◽  
Author(s):  
Junzo Hisatsune ◽  
Hideharu Hagiya ◽  
Sumiko Shiota ◽  
Motoyuki Sugai

ABSTRACT Staphylococcus aureus JH4899, a community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) isolate collected from a patient with systematically disseminated infection, is classified as sequence type 8 and carries the staphylococcal cassette chromosome mec type IVl (SCCmecIVl). It produces TSST-1, SEC, a newly discovered enterotoxin (SE1), and epidermal cell differentiation inhibitor A (EDIN-A). Here, we present the complete genome sequence of the chromosome and a plasmid harboring the se1 and ednA genes.


2021 ◽  
Vol 10 (37) ◽  
Author(s):  
Nicola M. Karakatsanis ◽  
Shakeel Mowlaboccus ◽  
Elena Colombi ◽  
Julie C. Pearson ◽  
Joshua P. Ramsay ◽  
...  

Sequence type 1 (ST1) methicillin-resistant Staphylococcus aureus (MRSA) type IV[2B] has become one of the most common community-associated MRSA clones in Australia. We report the complete genome sequence of one of the earliest isolated Australian S. aureus ST1-MRSA-IV strains, WBG8287, isolated from an Indigenous Australian patient living in the remote Kimberley Region of Western Australia.


2019 ◽  
Vol 57 (9) ◽  
Author(s):  
Ayesha Khan ◽  
Lina M. Rivas ◽  
Maria Spencer ◽  
Rodrigo Martinez ◽  
Marusella Lam ◽  
...  

ABSTRACT Ceftaroline (CPT) is a broad-spectrum agent with potent activity against methicillin-resistant Staphylococcus aureus (MRSA). The sequence type 5 (ST5) Chilean-Cordobés clone, associated with CPT nonsusceptibility, is dominant in Chile, a region with high rates of MRSA infections. Here, we assessed the in vitro activity of CPT against a collection of MRSA isolates collected between 1999 and 2018 from nine hospitals (n = 320) and community settings (n = 41) in Santiago, Chile, and evaluated performance across testing methodologies. We found that our hospital-associated isolates exhibited higher CPT MIC distributions (MIC50 and MIC90 of 2 mg/liter) than the community isolates (MIC50 and MIC90 of 0.5 mg/liter), a finding that was consistent across time and independent of the culture source. High proportions (64%) of isolates were CPT nonsusceptible despite the absence of CPT use in Chile. Across methodologies, the Etest underestimated the MIC relative to the gold standard broth microdilution (BMD) test (MIC50 and MIC90 of 1 and 1.5 mg/liter, respectively). There was low (∼51%) categorical agreement (CA) between Etest and BMD results across CLSI and EUCAST breakpoints. The recent revision of CLSI guidelines abolished “very major error” (VME) from the previous guidelines (81%), which perform similarly to the EUCAST guidelines. The level of concordance between CLSI and EUCAST for BMD testing and Etest was >95%. Disk diffusion performed poorly relative to BMD under CLSI (CA, 55%) and EUCAST (CA, 36%) guidelines. Comparison of EUCAST to CLSI for disk diffusion (with EUCAST used as the reference) showed low agreement (CA, 25%; VME, 70%). In summary, CPT-nonsusceptible MRSA are dominant in clinical settings in Chile. Our results provide data to support the reevaluation of CPT breakpoints and to improve agreement across methodologies and agencies.


2017 ◽  
Vol 5 (44) ◽  
Author(s):  
Samantha J. Hau ◽  
Darrell O. Bayles ◽  
David P. Alt ◽  
Timothy S. Frana ◽  
Tracy L. Nicholson

ABSTRACT Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) is a bacterium carried by or obtained from swine and other livestock. The initial and predominant swine-associated LA-MRSA sequence type (ST) identified is ST398. Here, we present 14 draft genome sequences from LA-MRSA ST398 isolates found in the United States.


mSystems ◽  
2021 ◽  
Author(s):  
Fangyou Yu ◽  
Astrid V. Cienfuegos-Gallet ◽  
Marcus H. Cunningham ◽  
Ye Jin ◽  
Bingjie Wang ◽  
...  

Staphylococcus aureus sequence type 9 (ST9) is the main LA-MRSA clone spreading in the Asian continent. It can colonize and cause mild to severe infections both in animal and humans.


2017 ◽  
Vol 5 (42) ◽  
Author(s):  
Samantha J. Hau ◽  
Darrell O. Bayles ◽  
David P. Alt ◽  
Tracy L. Nicholson

ABSTRACT Staphylococcus aureus is a commensal bacterium of humans that can cause a spectrum of diseases. An isolate’s capacity to cause disease is partially attributed to the acquisition of novel mobile genetic elements. This report provides the draft genome sequence of one methicillin-susceptible and seven methicillin-resistant clinical human S. aureus isolates.


2017 ◽  
Vol 5 (19) ◽  
Author(s):  
Olga Makarova ◽  
Paul Johnston ◽  
Birgit Walther ◽  
Jens Rolff ◽  
Uwe Roesler

ABSTRACT We report here the complete genome sequence of the livestock-associated methicillin-resistant Staphylococcus aureus strain 08S00974 from sequence type 398 (ST398 LA-MRSA) isolated from a fatting pig at a farm in Germany.


2015 ◽  
Vol 59 (6) ◽  
pp. 3669-3671 ◽  
Author(s):  
Jia Chang Cai ◽  
Yan Yan Hu ◽  
Hong Wei Zhou ◽  
Gong-Xiang Chen ◽  
Rong Zhang

ABSTRACTSixcfr-harboring methicillin-resistantStaphylococcus aureus(MRSA) isolates, which belonged to the same clone of sequence type 5 (ST5)-staphylococcal cassette chromosomemecelement II (SCCmecII)-spat311, were investigated in this study. Complete sequencing of acfr-carrying plasmid, pLRSA417, revealed an 8,487-bp fragment containing a Tn4001-like transposon,cfr,orf1, and ISEnfa4. This segment, first identified in an animal plasmid, pSS-01, was observed in several plasmids from clinical coagulase-negative staphylococci in China, suggesting that thecfrgene, which might originate from livestock, was located in the same mobile element and disseminated among different clinical staphylococcal species.


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