%0 Journal Article %1 honisch_automated_2007 %A Honisch, Christiane %A Chen, Yong %A Mortimer, Chloe %A Arnold, Catherine %A Schmidt, Oliver %A van den Boom, Dirk %A Cantor, Charles R %A Shah, Haroun N %A Gharbia, Saheer E %D 2007 %J Proceedings of the National Academy of Sciences of the United States of America %K Alleles, Analysis, Bacterial Bacterial, Base Cluster Data, Laser Mass, Molecular Neisseria Reproducibility Results, Sequence Sequence, Spectrometry, Techniques, Typing meningitidis, of {DNA,} {Desorption-Ionization} {Matrix-Assisted} %N 25 %P 10649--10654 %R 10.1073/pnas.0704152104 %T Automated comparative sequence analysis by base-specific cleavage and mass spectrometry for nucleic acid-based microbial typing %U http://www.ncbi.nlm.nih.gov/pubmed/17563374 %V 104 %X Traditional microbial typing technologies for the characterization of pathogenic microorganisms and monitoring of their global spread are often difficult to standardize and poorly portable, and they lack sufficient ease of use, throughput, and automation. To overcome these problems, we introduce the use of comparative sequencing by MALDI-TOF MS for automated high-throughput microbial DNA sequence analysis. Data derived from the public multilocus sequence typing (MLST) database (http://pubmlst.org/neisseria) established a reference set of expected peak patterns. A model pathogen, Neisseria meningitidis, was used to validate the technology and explore its applicability as an alternative to dideoxy sequencing. One hundred N. meningitidis samples were typed by comparing MALDI-TOF MS fingerprints of the standard MLST loci to reference sequences available in the public MLST database. Identification results can be obtained in 2 working days. Results were in concordance with classical dideoxy sequencing with 98\% correct automatic identification. Sequence types (STs) of 89 samples were represented in the database, seven samples revealed new STs, including three new alleles, and four samples contained mixed populations of multiple STs. The approach shows interlaboratory reproducibility and allows for the exchange of mass spectrometric fingerprints to study the geographic spread of epidemic N. meningitidis strains or other microbes of clinical importance.

@article{honisch_automated_2007, abstract = {Traditional microbial typing technologies for the characterization of pathogenic microorganisms and monitoring of their global spread are often difficult to standardize and poorly portable, and they lack sufficient ease of use, throughput, and automation. To overcome these problems, we introduce the use of comparative sequencing by {MALDI-TOF} {MS} for automated high-throughput microbial {DNA} sequence analysis. Data derived from the public multilocus sequence typing {(MLST)} database (http://pubmlst.org/neisseria) established a reference set of expected peak patterns. A model pathogen, Neisseria meningitidis, was used to validate the technology and explore its applicability as an alternative to dideoxy sequencing. One hundred N. meningitidis samples were typed by comparing {MALDI-TOF} {MS} fingerprints of the standard {MLST} loci to reference sequences available in the public {MLST} database. Identification results can be obtained in 2 working days. Results were in concordance with classical dideoxy sequencing with 98\% correct automatic identification. Sequence types {(STs)} of 89 samples were represented in the database, seven samples revealed new {STs,} including three new alleles, and four samples contained mixed populations of multiple {STs.} The approach shows interlaboratory reproducibility and allows for the exchange of mass spectrometric fingerprints to study the geographic spread of epidemic N. meningitidis strains or other microbes of clinical importance.}, added-at = {2011-03-11T10:05:34.000+0100}, author = {Honisch, Christiane and Chen, Yong and Mortimer, Chloe and Arnold, Catherine and Schmidt, Oliver and van den Boom, Dirk and Cantor, Charles R and Shah, Haroun N and Gharbia, Saheer E}, biburl = {https://www.bibsonomy.org/bibtex/2d8f511b76a9348bc53ebd2044d4bd745/jelias}, doi = {10.1073/pnas.0704152104}, interhash = {85a3a843e07bae825c0ba2a43d2cb6e0}, intrahash = {d8f511b76a9348bc53ebd2044d4bd745}, issn = {0027-8424}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, keywords = {Alleles, Analysis, Bacterial Bacterial, Base Cluster Data, Laser Mass, Molecular Neisseria Reproducibility Results, Sequence Sequence, Spectrometry, Techniques, Typing meningitidis, of {DNA,} {Desorption-Ionization} {Matrix-Assisted}}, month = jun, note = {{PMID:} 17563374}, number = 25, pages = {10649--10654}, timestamp = {2011-03-11T10:06:34.000+0100}, title = {Automated comparative sequence analysis by base-specific cleavage and mass spectrometry for nucleic acid-based microbial typing}, url = {http://www.ncbi.nlm.nih.gov/pubmed/17563374}, volume = 104, year = 2007 }