title: Multiplex Polymerase Chain Reaction for Identification of Shigellae and Four Shigella Species Using Novel Genetic Markers Screened by Comparative Genomics.
abstract: In the detection of Shigella species using molecular biological methods, previously known genetic markers for Shigella species were not sufficient to discriminate between Shigella species and diarrheagenic Escherichia coli. The purposes of this study were to screen for genetic markers of the Shigella genus and four Shigella species through comparative genomics and develop a multiplex polymerase chain reaction (PCR) for the detection of shigellae and Shigella species. A total of seven genomic DNA sequences from Shigella species were subjected to comparative genomics for the screening of genetic markers of shigellae and each Shigella species. The primer sets were designed from the screened genetic markers and evaluated using PCR with genomic DNAs from Shigella and other bacterial strains in Enterobacteriaceae. A novel Shigella quintuplex PCR, designed for the detection of Shigella genus, S. dysenteriae, S. boydii, S. flexneri, and S. sonnei, was developed from the evaluated primer sets, and its performance was demonstrated with specifically amplified results from each Shigella species. This Shigella multiplex PCR is the first to be reported with novel genetic markers developed through comparative genomics and may be a useful tool for the accurate detection of the Shigella genus and species from closely related bacteria in clinical microbiology and food safety.
authors.affiliation: Department of Food Science and Biotechnology, Kyung Hee University , Yongin, Republic of Korea.
authors.forename: Hyun-Joong
authors.initials: HJ
authors.lastname: Kim
authors.affiliation: Department of Food Science and Biotechnology, Kyung Hee University , Yongin, Republic of Korea.
authors.forename: Ji-Oh
authors.initials: JO
authors.lastname: Ryu
authors.affiliation: Department of Food Science and Biotechnology, Kyung Hee University , Yongin, Republic of Korea.
authors.forename: Ji-Yeon
authors.initials: JY
authors.lastname: Song
authors.affiliation: Department of Food Science and Biotechnology, Kyung Hee University , Yongin, Republic of Korea.
authors.forename: Hae-Yeong
authors.initials: HY
authors.lastname: Kim
mesh_terms: D055028:Comparative Genomic Hybridization; D017931:DNA Primers; D004269:DNA, Bacterial / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics* / Q000302:isolation & purification; D005819:Genetic Markers; D060885:Multiplex Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA; D012760:Shigella / Q000145:classification / Q000235:genetics* / Q000302:isolation & purification*
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D005819:Genetic Markers
keywords: comparative genomics; genetic marker; identification; multiplex PCR; shigellae