Characterization of Yersinia species by protein profiling using automated microfluidic capillary electrophoresis

Bennett, VM; McNevin, D; Roffey, P; Gahan, ME

Characterization of Yersinia species by protein profiling using automated microfluidic capillary electrophoresis

Bennett, VM McNevin, D

Roffey, P Gahan, ME

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Publication Type:: Journal Article
Citation:: Forensic Science, Medicine, and Pathology, 2017, 13 (1), pp. 10 - 19
Issue Date:: 2017-03-01

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Field	Value	Language
dc.contributor.author	Bennett, VM	en_US
dc.contributor.author	McNevin, D https://orcid.org/0000-0003-1665-3367	en_US
dc.contributor.author	Roffey, P	en_US
dc.contributor.author	Gahan, ME	en_US
dc.date.available	2016-11-07	en_US
dc.date.issued	2017-03-01	en_US
dc.identifier.citation	Forensic Science, Medicine, and Pathology, 2017, 13 (1), pp. 10 - 19	en_US
dc.identifier.issn	1547-769X	en_US
dc.identifier.uri	http://hdl.handle.net/10453/123717
dc.description.abstract	© 2016, Springer Science+Business Media New York. Yersinia pestis is a biological agent of high risk to national security due to its ability to be easily disseminated and transmitted among humans. If Y. pestis was to be utilized in a deliberate disease outbreak it would be essential to rapidly and accurately identify the agent. Current identification methods for Yersinia species are limited by their reliance on cultivation, the time taken to achieve results and/or the use of protocols that are not amenable for field use. Faster identification methods are urgently required. Microfluidic capillary electrophoresis was used to identify seven Yersinia species based on their protein profiles. Further objectives included determining if Yersinia species could be detected in mixtures of milk products and Escherichia coli, determining if Yersinia could be detected in a blinded identification and reproducibility across two platforms. Two characteristic protein bands were detected at 50 kilodaltons (kDa) and between 50 and 75 kDa for the Yersinia species. Individual Yersinia species could be differentiated from one another and distinguished from E. coli, Bacillus anthracis Sterne strain and Dipel (containing Bacillus thuringiensis). Due to the high protein content of milk products Yersinia could not be detected when mixed with these but was detected when mixed with E. coli. Species were correctly identified with 96% success in blinded procedures using 12 individuals. Whilst protein profile patterns were reproducible across platforms there was some discrepancy in protein sizing. This study demonstrates that protein profiling using microfluidic capillary electrophoresis is able to rapidly and reproducibly identify and characterize Yersinia species. Results show this technique is a powerful front-line, rapid and broad range screening method capable of identifying and differentiating biological agents, hoax agents and environmental bacterial species.	en_US
dc.relation.ispartof	Forensic Science, Medicine, and Pathology	en_US
dc.relation.isbasedon	10.1007/s12024-016-9824-7	en_US
dc.subject.classification	Legal & Forensic Medicine	en_US
dc.subject.mesh	Milk	en_US
dc.subject.mesh	Animals	en_US
dc.subject.mesh	Humans	en_US
dc.subject.mesh	Bacillus	en_US
dc.subject.mesh	Escherichia coli	en_US
dc.subject.mesh	Yersinia	en_US
dc.subject.mesh	Bacterial Proteins	en_US
dc.subject.mesh	Electrophoresis, Capillary	en_US
dc.subject.mesh	Reproducibility of Results	en_US
dc.subject.mesh	Microfluidics	en_US
dc.subject.mesh	Biological Warfare Agents	en_US
dc.title	Characterization of Yersinia species by protein profiling using automated microfluidic capillary electrophoresis	en_US
dc.type	Journal Article
utslib.citation.volume	1	en_US
utslib.citation.volume	13	en_US
utslib.for	1103 Clinical Sciences	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Faculty of Science/School of Mathematical and Physical Sciences
pubs.organisational-group	/University of Technology Sydney/Strength - CFS - Centre for Forensic Science
utslib.copyright.status	closed_access
pubs.issue	1	en_US
pubs.publication-status	Published	en_US
pubs.volume	13	en_US

Abstract:

© 2016, Springer Science+Business Media New York. Yersinia pestis is a biological agent of high risk to national security due to its ability to be easily disseminated and transmitted among humans. If Y. pestis was to be utilized in a deliberate disease outbreak it would be essential to rapidly and accurately identify the agent. Current identification methods for Yersinia species are limited by their reliance on cultivation, the time taken to achieve results and/or the use of protocols that are not amenable for field use. Faster identification methods are urgently required. Microfluidic capillary electrophoresis was used to identify seven Yersinia species based on their protein profiles. Further objectives included determining if Yersinia species could be detected in mixtures of milk products and Escherichia coli, determining if Yersinia could be detected in a blinded identification and reproducibility across two platforms. Two characteristic protein bands were detected at 50 kilodaltons (kDa) and between 50 and 75 kDa for the Yersinia species. Individual Yersinia species could be differentiated from one another and distinguished from E. coli, Bacillus anthracis Sterne strain and Dipel (containing Bacillus thuringiensis). Due to the high protein content of milk products Yersinia could not be detected when mixed with these but was detected when mixed with E. coli. Species were correctly identified with 96% success in blinded procedures using 12 individuals. Whilst protein profile patterns were reproducible across platforms there was some discrepancy in protein sizing. This study demonstrates that protein profiling using microfluidic capillary electrophoresis is able to rapidly and reproducibly identify and characterize Yersinia species. Results show this technique is a powerful front-line, rapid and broad range screening method capable of identifying and differentiating biological agents, hoax agents and environmental bacterial species.

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http://hdl.handle.net/10453/123717