Failure of phylogeny inferred from multilocus sequence typing to represent bacterial phylogeny

Although multilocus sequence typing (MLST) is highly discriminatory and useful for outbreak investigations and epidemiological surveillance, it has always been controversial whether clustering and phylogeny inferred from the MLST gene loci can represent the real phylogeny of bacterial strains. In this study, we compare the phylogenetic trees constructed using three approaches, (1) concatenated blocks of homologous sequence shared between the bacterial genomes, (2) genome single-nucleotide polymorphisms (SNP) profile and (3) concatenated nucleotide sequences of gene loci in the corresponding MLST schemes, for 10 bacterial species with >30 complete genome sequences available. Major differences in strain clustering at more than one position were observed between the phylogeny inferred using genome/SNP data and MLST for all 10 bacterial species. Shimodaira-Hasegawa test revealed significant difference between the topologies of the genome and MLST trees for nine of the 10 bacterial species, and significant difference between the topologies of the SNP and MLST trees were present for all 10 bacterial species. Matching Clusters and R-F Clusters metrics showed that the distances between the genome/SNP and MLST trees were larger than those between the SNP and genome trees. Phylogeny inferred from MLST failed to represent genome phylogeny with the same bacterial species.

[1]  Yi Chen,et al.  Implementation of Nationwide Real-time Whole-genome Sequencing to Enhance Listeriosis Outbreak Detection and Investigation. , 2016, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[2]  B. Spratt,et al.  Enhanced invasiveness of bovine-derived neonatal sequence type 17 group B streptococcus is independent of capsular serotype. , 2006, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[3]  Paul Keim,et al.  Phylogenetic discovery bias in Bacillus anthracis using single-nucleotide polymorphisms from whole-genome sequencing. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[4]  Jason W. Sahl,et al.  A Comparative Genomic Analysis of Diverse Clonal Types of Enterotoxigenic Escherichia coli Reveals Pathovar-Specific Conservation , 2010, Infection and Immunity.

[5]  Paul Keim,et al.  Anthrax molecular epidemiology and forensics: using the appropriate marker for different evolutionary scales. , 2004, Infection, genetics and evolution : journal of molecular epidemiology and evolutionary genetics in infectious diseases.

[6]  B. Li,et al.  Genotypic Analysis of Klebsiella pneumoniae Isolates in a Beijing Hospital Reveals High Genetic Diversity and Clonal Population Structure of Drug-Resistant Isolates , 2013, PloS one.

[7]  P. Woo,et al.  Evidence for Elizabethkingia anophelis Transmission from Mother to Infant, Hong Kong , 2015, Emerging infectious diseases.

[8]  A. Nekrutenko,et al.  Galaxy: a comprehensive approach for supporting accessible, reproducible, and transparent computational research in the life sciences , 2010, Genome Biology.

[9]  Thomas J Naughton,et al.  Assessment of methods for amino acid matrix selection and their use on empirical data shows that ad hoc assumptions for choice of matrix are not justified , 2006, BMC Evolutionary Biology.

[10]  M. Achtman,et al.  Multilocus sequence typing: a portable approach to the identification of clones within populations of pathogenic microorganisms. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[11]  R. Goering,et al.  Molecular Epidemiology of Methicillin-Resistant and Methicillin-Susceptible Staphylococcus aureus Isolates from Global Clinical Trials , 2008, Journal of Clinical Microbiology.

[12]  Dag Harmsen,et al.  Defining and Evaluating a Core Genome Multilocus Sequence Typing Scheme for Whole-Genome Sequence-Based Typing of Listeria monocytogenes , 2015, Journal of Clinical Microbiology.

[13]  B. Cookson,et al.  Polyclonal multiply antibiotic-resistant methicillin-resistant Staphylococcus aureus with Panton-Valentine leucocidin in England. , 2010, The Journal of antimicrobial chemotherapy.

[14]  Eugene W. Myers,et al.  Basic local alignment search tool. Journal of Molecular Biology , 1990 .

[15]  Daniel J. Blankenberg,et al.  Galaxy: A Web‐Based Genome Analysis Tool for Experimentalists , 2010, Current protocols in molecular biology.

[16]  M. Maiden,et al.  Multilocus Sequence Typing System forCampylobacter jejuni , 2001, Journal of Clinical Microbiology.

[17]  Brian G. Spratt,et al.  Multilocus Sequence Typing of Streptococcus pyogenes and the Relationships between emm Type and Clone , 2001, Infection and Immunity.

[18]  C. Robert,et al.  Clostridium butyricum Strains and Dysbiosis Linked to Necrotizing Enterocolitis in Preterm Neonates. , 2015, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[19]  D. W. Kim,et al.  Shigella sonnei genome sequencing and phylogenetic analysis indicate recent global dissemination from Europe , 2012, Nature Genetics.

[20]  N. Loman,et al.  High-throughput bacterial genome sequencing: an embarrassment of choice, a world of opportunity , 2012, Nature Reviews Microbiology.

[21]  O. Gascuel,et al.  A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. , 2003, Systematic biology.

[22]  Julian Parkhill,et al.  Evolution of MRSA During Hospital Transmission and Intercontinental Spread , 2010, Science.

[23]  C. Rodrigues,et al.  Molecular Characterization of Methicillin-Resistant Staphylococcus aureus with Emergence of Epidemic Clones of Sequence Type (ST) 22 and ST 772 in Mumbai, India , 2010, Journal of Clinical Microbiology.

[24]  F. Rousset,et al.  Emergence and dissemination of extended-spectrum beta-lactamase-producing Escherichia coli in the community: lessons from the study of a remote and controlled population. , 2010, The Journal of infectious diseases.

[25]  Nicholas P. J. Day,et al.  Multilocus Sequence Typing for Characterization of Methicillin-Resistant and Methicillin-Susceptible Clones ofStaphylococcus aureus , 2000, Journal of Clinical Microbiology.

[26]  Brian D. Ondov,et al.  The Harvest suite for rapid core-genome alignment and visualization of thousands of intraspecific microbial genomes , 2014, Genome Biology.

[27]  Paramvir S. Dehal,et al.  FastTree 2 – Approximately Maximum-Likelihood Trees for Large Alignments , 2010, PloS one.

[28]  Steven Salzberg,et al.  Mugsy: fast multiple alignment of closely related whole genomes , 2010, Bioinform..

[29]  B. Spratt,et al.  A multilocus sequence typing scheme for Streptococcus pneumoniae: identification of clones associated with serious invasive disease. , 1998, Microbiology.

[30]  P. Woo,et al.  A novel MLST sequence type discovered in the first fatal case of Laribacter hongkongensis bacteremia clusters with the sequence types of other human isolates , 2014, Emerging Microbes & Infections.

[31]  P. Woo,et al.  Analysis of multilocus sequence typing schemes for 35 different bacteria revealed that gene loci of 10 bacteria could be replaced to improve cost-effectiveness. , 2011, Diagnostic microbiology and infectious disease.

[32]  H. Davies,et al.  Multilocus sequence typing of serotype III group B streptococcus and correlation with pathogenic potential. , 2004, The Journal of infectious diseases.

[33]  A. van Belkum,et al.  Prevalence of Panton-Valentine leukocidin genes among carriage and invasive Staphylococcus aureus isolates in Malaysia. , 2009, International journal of infectious diseases : IJID : official publication of the International Society for Infectious Diseases.

[34]  Keith A. Jolley,et al.  Ribosomal multilocus sequence typing: universal characterization of bacteria from domain to strain , 2012, Microbiology.

[35]  Rodrigo Lopez,et al.  Clustal W and Clustal X version 2.0 , 2007, Bioinform..

[36]  Daniel J. Blankenberg,et al.  Galaxy: a platform for interactive large-scale genome analysis. , 2005, Genome research.

[37]  A. van Belkum,et al.  Associations between Staphylococcus aureus Genotype, Infection, and In-Hospital Mortality: A Nested Case-Control Study. , 2005, The Journal of infectious diseases.

[38]  Wei Qian,et al.  Selection of conserved blocks from multiple alignments for their use in phylogenetic analysis. , 2000, Molecular biology and evolution.

[39]  P. Woo,et al.  Development of a multi-locus sequence typing scheme for Laribacter hongkongensis, a novel bacterium associated with freshwater fish-borne gastroenteritis and traveler's diarrhea , 2009, BMC Microbiology.

[40]  Lori A. S. Snyder,et al.  The Complete Genome and Proteome of Laribacter hongkongensis Reveal Potential Mechanisms for Adaptations to Different Temperatures and Habitats , 2009, PLoS genetics.

[41]  J. Bray,et al.  MLST revisited: the gene-by-gene approach to bacterial genomics , 2013, Nature Reviews Microbiology.

[42]  F. Baquero,et al.  Molecular analysis of community-acquired methicillin-susceptible and resistant Staphylococcus aureus isolates recovered from bacteraemic and osteomyelitis infections in children from Tunisia. , 2011, Clinical microbiology and infection : the official publication of the European Society of Clinical Microbiology and Infectious Diseases.

[43]  Hidetoshi Shimodaira,et al.  Multiple Comparisons of Log-Likelihoods with Applications to Phylogenetic Inference , 1999, Molecular Biology and Evolution.

[44]  E. Myers,et al.  Basic local alignment search tool. , 1990, Journal of molecular biology.

[45]  B. Spratt,et al.  Multilocus sequence typing for characterization of methicillin-resistant and methicillin-susceptible clones of Staphylococcus aureus. , 2000, Journal of clinical microbiology.

[46]  M. Nei,et al.  MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. , 2007, Molecular biology and evolution.

[47]  D. Swofford PAUP*: Phylogenetic analysis using parsimony (*and other methods), Version 4.0b10 , 2002 .

[48]  Savitri Sharma,et al.  Staphylococcus aureus eye infections in two Indian hospitals: emergence of ST772 as a major clone , 2012, Clinical ophthalmology.