Whole-genome sequencing for routine pathogen surveillance in public health : A population snapshot of invasive Staphylococcus aureus in Europe

Funding Information: This work, including the efforts of Matthew Holden, Janina Dordel, Julian Parkhill, and Stephen Bentley, was funded by Wellcome Trust (098051). This work, including the efforts of David M. Aanensen, Corin Yeats, and Artemij Fedosejev, was funded by Wellcome Trust (099202). This work, including the efforts of Brian Spratt, was funded by Wellcome Trust (089472). This work, including the efforts of Santiago Castillo-Ram?rez, was funded by Medical Research Council (MRC) (G1000803). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Publisher Copyright: © 2016 Aanensen et al.The implementation of routine whole-genome sequencing (WGS) promises to transform our ability to monitor the emergence and spread of bacterial pathogens. Here we combined WGS data from 308 invasive Staphylococcus aureus isolates corresponding to a pan-European population snapshot, with epidemiological and resistance data. Geospatial visualization of the data is made possible by a generic software tool designed for public health purposes that is available at the project URL (http:// www.microreact.org/project/EkUvg9uY?tt=rc). Our analysis demonstrates that high-risk clones can be identified on the basis of population level properties such as clonal relatedness, abundance, and spatial structuring and by inferring virulence and resistance properties on the basis of gene content. We also show that in silico predictions of antibiotic resistance profiles are at least as reliable as phenotypic testing. We argue that this work provides a comprehensive road map illustrating the three vital components for future molecular epidemiological surveillance: (i) large-scale structured surveys, (ii) WGS, and (iii) communityoriented database infrastructure and analysis tools. IMPORTANCE The spread of antibiotic-resistant bacteria is a public health emergency of global concern, threatening medical intervention at every level of health care delivery. Several recent studies have demonstrated the promise of routine wholegenome sequencing (WGS) of bacterial pathogens for epidemiological surveillance, outbreak detection, and infection control. However, as this technology becomes more widely adopted, the key challenges of generating representative national and international data sets and the development of bioinformatic tools to manage and interpret the data become increasingly pertinent. This study provides a road map for the integration of WGS data into routine pathogen surveillance. We emphasize the importance of large-scale routine surveys to provide the population context for more targeted or localized investigation and the development of open-access bioinformatic tools to provide the means to combine and compare independently generated data with publicly available data sets.publishersversionPeer reviewe

Analysis of a database of DNA profiles of 734 hybrid tea rose varieties

Over 10,000 Hybrid Tea rose varieties have been described. The large number of varieties and the size of the reference collections may cause problems in DUS (Distinctiveness, Uniformity, and Stability) testing. Molecular markers may help to solve these problems by providing high power to identify and recognise seedling-derived varieties based on unique genotypes, while grouping mutants into groups with identical marker scores. Here we describe the use of a set of 11 rose microsatellite markers to generate a database of molecular profiles of Hybrid Tea varieties. The data were analysed with respect to reproducibility, discriminative power, genetic (sub) structure, and correlation between molecular and DUS characteristics. The use of the markers in the DUS context is discussed with respect to the options 2 and 3 as they were formulated by the UPOV-BMT working group. It is concluded that an option 3 approach (granting of PBR based on distinctness observed with molecular markers) is feasible for ros

Inducers of epithelial mesenchymal transition and cancer stem cells in malignant pleural effusions

The Epithelial to Mesenchymal Transition (EMT) plays a role not only in tumor metastasis but also in tumor recurrence. This process is believed to be tightly linked to the presence of Cancer Stem Cells (CSCs) however, it is still not clear which factors could induce EMT and how it could be a source for CSCs. It has been demonstrated that Malignant Pleural Effusion (MPEs) may represent an excellent source to identify markers and molecular mechanisms involved in EMT and CSCs development. Growth factors, cell differentiation markers and molecular adhesion are involved in some of the crucial neoplastic cell events such as proliferation, metastasis, resistance to chemotherapy and EMT. In this review, we summarize the current understanding of which molecular markers can orchestrate EMT and CSCs in MPEs

Marker development in ornamental plants

Development of markers for a new crop or development of additional markers for a crop where markers have been developed in the past raises the question of the intended use of the markers. Depending on the different objectives in mind one marker type may be better suited then another. In general one can think of two main objectives for the use of markers; variety identification and breeding applications. In view of recent developments in molecular genetics, and sequencing technologies in particular, within the 23rd International Eucarpia Symposium Section Ornamentals a workshop was devoted on molecular markers and their use in ornamentals. Within this paper an overview will be presented on the development of markers for identification of ornamental crops and on the importance of the new developments in marker and sequence technology for the use of markers in ornamental breedin

On the informativeness of dominant and co-dominant genetic markers for Bayesian supervised clustering

We study the accuracy of Bayesian supervised method used to cluster individuals into genetically homogeneous groups on the basis of dominant or codominant molecular markers. We provide a formula relating an error criterion the number of loci used and the number of clusters. This formula is exact and holds for arbitrary number of clusters and markers. Our work suggests that dominant markers studies can achieve an accuracy similar to that of codominant markers studies if the number of markers used in the former is about 1.7 times larger than in the latter

Variable number of tandem repeat markers in the genome sequence of Mycosphaerella fijiensis, the causal agent of black leaf streak disease of banana (Musa spp)

ABSTRACT. We searched the genome of Mycosphaerella fijiensis for molecular markers that would allow population genetics analysis of this plant pathogen. M. fijiensis, the causal agent of banana leaf streak disease, also known as black Sigatoka, is the most devastating pathogen attacking bananas (Musa spp). Recently, the entire genome sequence of M. fijiensis became available. We screened this database for VNTR markers. Forty-two primer pairs were selected for validation, based on repeat type and length and the number of repeat units. Five VNTR markers showing multiple alleles were validated with a reference set of isolates from different parts of the world and a population from a banana plantation in Costa Rica. Polymorphism information content values varied from 0.6414 to 0.7544 for the reference set and from 0.0400 and 0.7373 for the population set. Eighty percent of the polymorphism information content values were above 0.60, indicating that the markers are highly informative. These markers allowed robust scoring of agarose gels and proved to be useful for variability and population genetics studies. In conclusion, the strategy we developed to identify and validate VNTR markers is an efficient means to incorporate markers that can be used for fungicide resistance management and to develop breeding strategies to control banana black leaf streak disease. This is the first report of VNTR-minisatellites from the M. fijiensis genome sequence. Key words: Molecular markers; VNTRs; Genetic diversity; Population genetics; Black Sigatok