Richard R Chapleau1, James C Baldwin1. 1. Applied Technology and Genomics Center, United States Air Force School of Aerospace Medicine , 711th Human Performance Wing, Air Force Research Laboratory, Wright-Patterson AFB OH .
Abstract
INTRODUCTION: Optical restriction genome mapping is a technology in which a genome is linearized on a surface and digested with specific restriction enzymes, giving an arrangement of the genome with gaps whose order and size are unique for a given organism. Current applications of this technology include assisting with the correct scaffolding and ordering of genomes in conjunction with whole-genome sequencing, observation of genetic drift and evolution using comparative genomics and epidemiological monitoring of the spread of infections. Here, we investigated the suitability of genome mapping for use in clinical labs as a potential diagnostic tool. MATERIALS AND METHODS: Using whole genome mapping, we investigated the basic performance of the technology for identifying two bacteria of interest for food-safety (Lactobacilli spp. and Enterohemorrhagic Escherichia coli). We further evaluated the performance for identifying multiple organisms from both simple and complex mixtures. RESULTS: We were able to successfully generate optical restriction maps of four Lactobacillus species as well as a strain of Enterohemorrhagic Escherichia coli from within a mixed solution, each distinguished using a common compatible restriction enzyme. Finally, we demonstrated that optical restriction maps were successfully obtained and the correct organism identified within a clinical matrix. CONCLUSION: With additional development, whole genome mapping may be a useful clinical tool for rapid invitro diagnostics.
INTRODUCTION: Optical restriction genome mapping is a technology in which a genome is linearized on a surface and digested with specific restriction enzymes, giving an arrangement of the genome with gaps whose order and size are unique for a given organism. Current applications of this technology include assisting with the correct scaffolding and ordering of genomes in conjunction with whole-genome sequencing, observation of genetic drift and evolution using comparative genomics and epidemiological monitoring of the spread of infections. Here, we investigated the suitability of genome mapping for use in clinical labs as a potential diagnostic tool. MATERIALS AND METHODS: Using whole genome mapping, we investigated the basic performance of the technology for identifying two bacteria of interest for food-safety (Lactobacilli spp. and Enterohemorrhagic Escherichia coli). We further evaluated the performance for identifying multiple organisms from both simple and complex mixtures. RESULTS: We were able to successfully generate optical restriction maps of four Lactobacillus species as well as a strain of Enterohemorrhagic Escherichia coli from within a mixed solution, each distinguished using a common compatible restriction enzyme. Finally, we demonstrated that optical restriction maps were successfully obtained and the correct organism identified within a clinical matrix. CONCLUSION: With additional development, whole genome mapping may be a useful clinical tool for rapid invitro diagnostics.
Authors: Scott A Jackson; Michael L Kotewicz; Isha R Patel; David W Lacher; Jayanthi Gangiredla; Christopher A Elkins Journal: Appl Environ Microbiol Date: 2011-12-30 Impact factor: 4.792
Authors: Paul D R Johnson; Susan A Ballard; Elizabeth A Grabsch; Timothy P Stinear; Torsten Seemann; Heather L Young; M Lindsay Grayson; Benjamin P Howden Journal: J Infect Dis Date: 2010-10-15 Impact factor: 5.226
Authors: Randi Føns Petersen; Eva Litrup; Jonas T Larsson; Mia Torpdahl; Gitte Sørensen; Luise Müller; Eva M Nielsen Journal: Foodborne Pathog Dis Date: 2011-03-07 Impact factor: 3.171
Authors: Matthew C Riley; Benjamin C Kirkup; Jake D Johnson; Emil P Lesho; Christian F Ockenhouse Journal: Malar J Date: 2011-08-26 Impact factor: 2.979
Authors: Fatma Onmus-Leone; Jun Hang; Robert J Clifford; Yu Yang; Matthew C Riley; Robert A Kuschner; Paige E Waterman; Emil P Lesho Journal: PLoS One Date: 2013-04-17 Impact factor: 3.240
Authors: Gene E Ananiev; Steve Goldstein; Rod Runnheim; Dan K Forrest; Shiguo Zhou; Konstantinos Potamousis; Chris P Churas; Veit Bergendahl; James A Thomson; David C Schwartz Journal: BMC Mol Biol Date: 2008-07-30 Impact factor: 2.946
Authors: Colin Raeside; Joël Gaffé; Daniel E Deatherage; Olivier Tenaillon; Adam M Briska; Ryan N Ptashkin; Stéphane Cruveiller; Claudine Médigue; Richard E Lenski; Jeffrey E Barrick; Dominique Schneider Journal: mBio Date: 2014-09-09 Impact factor: 7.867