title: Combination of biobarcode assay with on-chip capillary electrophoresis for ultrasensitive and multiplex biological agent detection.
abstract: Early diagnosis of biological agents is of paramount importance to prevent the casualties and fatal disease in human during bioterrorism or biological warfare. In this study, we reported an efficient and sensitive multiplex biological agent detection method based on the DNA biobarcode assay and the micro-capillary electrophoresis (μCE) technology. Monoplex as well as multiplex pathogen identification was performed using five targets including Bacillus anthracis, Francisella tularensis, Yersinia pestis, Vaccinia virus and Botulinum toxin A. Through the DNA biobarcode assay process, the magnetic microparticle-pathogen-polystyrene microbead complexes were formed, and the FAM labeled single stranded barcode DNA could be released from the complexes upon denaturation. Different lengths of a barcode DNA were designed to designate each pathogen, so that the specific peak elution time in the capillary electrophoresis on a chip allows us to distinguish the target with high accuracy within 3 min. We improved the assignment accuracy of the peak in the electropherogram by adding two bracket ladders. Owing to the abundant amount of barcode DNAs, the presence of B. anthracis, F. tularensis, Y. pestis, Vaccinia virus was confirmed with a limit of detection of 50CFU/mL, while Botulinum toxin A was analyzed even at a concentration of 12.5 ag/mL. Multiple pathogen detection was also successfully conducted in a phosphate buffered saline (PBS) as well as a serum medium with background of other pathogens. Thus, our analytical platform based on the biobarcode assay and on-chip CE analysis provides rapid, sensitive, multiplex, and accurate biological agent identification.
authors.affiliation: Department of Chemical and Biomolecular Engineering (BK21 PLUS Program), Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea. Electronic address: mkcho25@kaist.ac.kr.
authors.forename: Minkyung
authors.initials: M
authors.lastname: Cho
authors.affiliation: Department of Chemical and Biomolecular Engineering (BK21 PLUS Program), Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea. Electronic address: csy717@kaist.ac.kr.
authors.forename: Soyi
authors.initials: S
authors.lastname: Chung
authors.affiliation: Department of Chemical and Biomolecular Engineering (BK21 PLUS Program), Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea. Electronic address: jjaehwan@kaist.ac.kr.
authors.forename: Jae Hwan
authors.initials: JH
authors.lastname: Jung
authors.affiliation: Division of High-Risk Pathogen Research, Center for Infectious Diseases, National Institute of Health, 187 Osongsaengmyeong 2-ro, Cheongwon-gun, Chungbuk 363-951, Republic of Korea. Electronic address: gerhie@nih.go.kr.
authors.forename: Gi-eun
authors.initials: GE
authors.lastname: Rhie
authors.affiliation: Division of High-Risk Pathogen Research, Center for Infectious Diseases, National Institute of Health, 187 Osongsaengmyeong 2-ro, Cheongwon-gun, Chungbuk 363-951, Republic of Korea. Electronic address: jhjeon78@korea.kr.
authors.forename: Jun Ho
authors.initials: JH
authors.lastname: Jeon
authors.affiliation: Department of Chemical and Biomolecular Engineering (BK21 PLUS Program), Korea Advanced Institute of Science and Technology (KAIST), 291 Daehak-ro, Yuseong-gu, Daejeon 305-701, Republic of Korea. Electronic address: seots@kaist.ac.kr.
authors.forename: Tae Seok
authors.initials: TS
authors.lastname: Seo
mesh_terms: D001408:Bacillus anthracis / Q000235:genetics / Q000302:isolation & purification; D001424:Bacterial Infections / Q000175:diagnosis / Q000382:microbiology; D015374:Biosensing Techniques / Q000295:instrumentation*; D023482:Bioterrorism; D019274:Botulinum Toxins, Type A / Q000235:genetics / Q000302:isolation & purification; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification*; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification*; D019075:Electrophoresis, Capillary / Q000295:instrumentation*; D004867:Equipment Design; D005604:Francisella tularensis / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D014615:Vaccinia / Q000175:diagnosis / Q000235:genetics; D014616:Vaccinia virus / Q000235:genetics / Q000302:isolation & purification; D015010:Yersinia pestis / Q000235:genetics / Q000302:isolation & purification
chemical_list: D004269:DNA, Bacterial; D004279:DNA, Viral; D019274:Botulinum Toxins, Type A
keywords: Barcode assay; Biological agents; Capillary electrophoresis; Chip; Pathogen detection