title: Designing multiplex PCR system of Campylobacter jejuni for efficient typing by improving monoplex PCR binary typing method.
abstract: Campylobacter jejuni is responsible for the majority of Campylobacter infections. As the molecular epidemiological study of outbreaks, pulsed-field gel electrophoresis (PFGE) is performed in general. But PFGE has several problems. PCR binary typing (P-BIT) method is a typing method for Campylobacter spp. that was recently developed, and was reported to have a similar discriminatory power and stability to those of PFGE. We modified the P-BIT method from 18 monoplex PCRs to two multiplex PCR systems (mP-BIT). The same results were obtained from monoplex PCRs using original primers and multiplex PCR in the representative isolates. The mP-BIT can analyze 48 strains at a time by using 96-well PCR systems and can identify C. jejuni because mP-BIT includes C. jejuni marker. The typing of the isolates by the mP-BIT and PFGE demonstrated generally concordant results and the mP-BIT method (D = 0.980) has a similar discriminatory power to that of PFGE with SmaI digest (D = 0.975) or KpnI digest (D = 0.987) as with original article. The mP-BIT method is quick, simple and easy, and comes to be able to perform it at low cost by having become a multiplex PCR system. Therefore, the mP-BIT method with two multiplex PCR systems has high potential for a rapid first-line surveillance typing assay of C. jejuni and can be used for routine surveillance and outbreak investigations of C. jejuni in the future.
authors.affiliation: Department of Microbiology and Medical Zoology, Aichi Prefectural Institute of Public Health, Nagoya 462-8576, Japan; Department of Biological Chemistry, College of Bioscience and Biotechnology, Chubu University, Kasugai, Aichi 487-8501, Japan. Electronic address: kazuhiro_yamada@pref.aichi.lg.jp.
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Yamada
authors.affiliation: Department of Microbiology and Medical Zoology, Aichi Prefectural Institute of Public Health, Nagoya 462-8576, Japan.
authors.forename: Ami
authors.initials: A
authors.lastname: Ibata
authors.affiliation: Department of Microbiology and Medical Zoology, Aichi Prefectural Institute of Public Health, Nagoya 462-8576, Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Suzuki
authors.affiliation: Department of Microbiology and Medical Zoology, Aichi Prefectural Institute of Public Health, Nagoya 462-8576, Japan.
authors.forename: Masakado
authors.initials: M
authors.lastname: Matsumoto
authors.affiliation: Department of Microbiology and Medical Zoology, Aichi Prefectural Institute of Public Health, Nagoya 462-8576, Japan.
authors.forename: Teruo
authors.initials: T
authors.lastname: Yamashita
authors.affiliation: Department of Microbiology and Medical Zoology, Aichi Prefectural Institute of Public Health, Nagoya 462-8576, Japan.
authors.forename: Hiroko
authors.initials: H
authors.lastname: Minagawa
authors.affiliation: Department of Biological Chemistry, College of Bioscience and Biotechnology, Chubu University, Kasugai, Aichi 487-8501, Japan.
authors.forename: Ryuichiro
authors.initials: R
authors.lastname: Kurane
mesh_terms: D002169:Campylobacter Infections / Q000382:microbiology*; D016123:Campylobacter jejuni / Q000145:classification / Q000235:genetics*; D016000:Cluster Analysis; D016521:Electrophoresis, Gel, Pulsed-Field; D006801:Humans; D058889:Molecular Typing / Q000379:methods*; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D010802:Phylogeny
keywords: Campylobacter jejuni; PCR binary typing; Pulsed-field gel electrophoresis (PFGE)