Multilocus sequence typing

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Multilocus sequence typing (to German multi- locus -Sequenztypisierung , MLST) is a biochemical and bioinformatic method of Phylogenomik to determine degrees of relationship between species and subspecies, especially prokaryotes . The MLST was first performed in 1998 by Martin Maiden and colleagues on Neisseria meningitidis .

properties

To get a clearer result, a clone is usually isolated by smear . A DNA extraction is carried out on the clone . The MLST uses the polymerase chain reaction (PCR) to propagate at least five to seven household genes , followed by DNA sequencing of 400 to 500 base pairs of the household genes. A number is assigned to each distinguishable sequence of one of the household genes examined. The combination of these figures results in a sequence type ( English sequence type ) and is used to identify and distinguish strains. The MLST can also be combined with the Multilocus Sequence Analysis (MLSA), in which only the differing household genes from a previously performed MLST are examined.

Due to the variability of household genes, the MLST is only suitable for distinguishing closely related species and strains. The strains of some species cannot be differentiated using the MLST, which is why additional typifications are used - the test strength of the MLST is then insufficient. The MLST is often used in addition to sequencing the 16S rDNA or for sequencing tandem repeats . When differentiating between pathogenic and non-pathogenic bacterial strains, a variant of the MLST is used, the MVLST ( English Multi-virulence-locus sequence typing ). The MVLST examines the genes of virulence factors instead of household genes , which differ more strongly in comparison and also enable statements to be made about the course of the disease . Genome sequencing offers the maximum distinctness (but also higher costs) .

Online databases for MLST are PubMLST and Institut Pasteur MLST . The Bacterial Isolate Genome Sequence Database (BIGSdb) collects MLST from genome sequencing.

Individual evidence

  1. ↑ a b Michael Goodfellow, Iain Sutcliffe, Jongsik Chun: New Approaches to Prokaryotic Systematics. ISBN 0128001763 . P. 221.
  2. ↑ MC Maiden, JA Bygraves, E. Feil, G. Morelli, JE Russell, R. Urwin, Q. Zhang, J. Zhou, K. Zurth, DA Caugant, IM Feavers, M. Achtman, BG Spratt: Multilocus sequence typing : a portable approach to the identification of clones within populations of pathogenic microorganisms. In: Proceedings of the National Academy of Sciences . Volume 95, number 6, March 1998, pp. 3140-3145, doi : 10.1073 / pnas.95.6.3140 , PMID 9501229 , PMC 19708 (free full text).
  3. ↑ a b c Rainer Kurmayer, Kaarina Sivonen, Annick Wilmotte, Nico Salmaso: Molecular Tools for the Detection and Quantification of Toxigenic Cyanobacteria. ISBN 978-1-119-33210-7 ( limited preview in Google Book Search).
  4. ↑ a b M. C. Maiden, MJ Jansen van Rensburg, JE Bray, SG Earle, SA Ford, KA Jolley, ND McCarthy: MLST revisited: the gene-by-gene approach to bacterial genomics. In: Nature reviews. Microbiology. Volume 11, number 10, 10 2013, pp. 728-736, doi : 10.1038 / nrmicro3093 , PMID 23979428 , PMC 3980634 (free full text).
  5. ↑ DM Adair, PL Worsham, KK Hill, AM Klevytska, PJ Jackson, AM Friedlander, P. Keim: Diversity in a variable-number tandem repeat from Yersinia pestis. In: Journal of clinical microbiology. Volume 38, Number 4, April 2000, pp. 1516-1519, PMID 10747136 , PMC 86479 (free full text).
  6. ↑ W. Zhang, BM Jayarao, SJ Knabel: Multi-virulence locus sequence typing of Listeria monocytogenes. In: Applied and Environmental Microbiology. Volume 70, number 2, February 2004, pp. 913-920, doi : 10.1128 / aem.70.2.913-920.2004 , PMID 14766571 , PMC 348834 (free full text).
  7. ↑ KA Jolley, JE Bray, MC Maiden: Open-access bacterial population genomics: BIGSdb software, the PubMLST.org website and their applications. In: Wellcome open research. Volume 3, 2018, p. 124, doi : 10.12688 / wellcomeopenres.14826.1 , PMID 30345391 , PMC 6192448 (free full text).
  8. ↑ Institut Pasteur : Institut Pasteur MLST , accessed on December 19, 2019.
  9. ^ KA Jolley, MC Maiden: BIGSdb: Scalable analysis of bacterial genome variation at the population level. In: BMC Bioinformatics. Volume 11, December 2010, p. 595, doi : 10.1186 / 1471-2105-11-595 , PMID 21143983 , PMC 3004885 (free full text).