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|Title:||Translational Genomics to Develop a Salmonella enterica Serovar Paratyphi A Multiplex Polymerase Chain Reaction Assay.|
Ju, Cindy Teh Shuan
Barer, Michael R.
|Publisher:||The American Society for Investigative Pathology and the Association for Molecular Pathology|
|Citation:||Journal of Molecular Diagnostics, 2007, 9 (5), pp. 624-630.|
|Abstract:||The use of pathogen genome sequence data for the control and management of infections remains an ongoing challenge. We describe a broadly applicable, web-enabled approach that can be used to develop bacteria-specific polymerase chain reaction (PCR) assays. Salmonella enterica Paratyphi A has emerged as a major cause of enteric fever in Asia. Culture-based diagnosis is slow and frequently negative in patients with suspected typhoid and paratyphoid fever, potentially compromising patient management and public health. We used the MobilomeFINDER web-server to perform in silico subtractive hybridization, thus identifying 43 protein-coding sequences (CDSs) that were present in two Paratyphi A strains but not in other sequenced Salmonella genomes. After exclusion of 29 CDSs found to be variably present in Paratyphi A strains by microarray hybridization and grouping of remaining CDSs by genomic location, four dispersed targets (stkF, spa2473, spa2539, hsdM) were used to develop a highly discriminatory multiplex PCR assay. All 52 Paratyphi A strains within the diverse panel investigated produced one of two athognomonic four-band signatures. Given rapid and ongoing expansion of DNA and comparative genomics databases, our universally accessible web-server-supported do-it-yourself approach offers the potential to contribute significantly to the rapid development of species-, serovar-, or pathotype-specific PCR assays targeting pre-existing and emerging bacterial pathogens.|
|Rights:||This paper was published as Journal of Molecular Diagnostics, 2007, 9 (5), pp. 624-630. Copyright © The American Society for Investigative Pathology and the Association for Molecular Pathology. Doi: 10.2353/jmoldx.2007.070064. Reprinted from The Journal of Molecular Diagnostics with permission from the American Society for Investigative Pathology and the Association for Molecular Pathology. Any reproduction or reuse of this material including but not limited to reformatting, reposting, republication, translation, or other derivative works based on any portion of this article, will require separate permission from the Publisher (ASIP). Definitive version of the article appears on http://jmd.amjpathol.org/.|
|Appears in Collections:||Published Articles, Dept. of Infection, Immunity and Inflammation|
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