title: Novel multiplex polymerase chain reaction primer set for identification of Lactococcus species.
abstract: OBJECTIVE
  The gram-positive bacterial genus Lactococcus has been taxonomically classified into seven species (Lactococcus lactis, Lactococcus garvieae, Lactococcus piscium, Lactococcus plantarum, Lactococcus raffinolactis, Lactococcus chungangensis and Lactococcus fujiensis). This study aimed to develop a novel multiplex polymerase chain reaction (PCR) primer set for the identification of the seven lactococcal species, as well as to differentiate the two industrially important dairy subspecies, L. lactis subsp. lactis and L. lactis subsp. cremoris.
  
  
  RESULTS
  A multiplex PCR primer set was designed based on the nucleotide sequences of the 16S rRNA gene of the seven lactococcal species. The specificity of the established one-step multiplex PCR scheme was verified using more than 200 bacterial strains, in which a complete sequence match was confirmed by partial sequencing of their 16S rRNA gene.
  
  
  CONCLUSIONS
  The one-step multiplex PCR enables the identification and speciation of bacterial strains belonging to the genus Lactococcus and the differentiation of strains of L. lactis subsp. lactis and L. lactis subsp. cremoris.
  
  
  CONCLUSIONS
  This work provides an efficient method for identification of lactococcal strains of industrial importance.
authors.affiliation: Food Science and Technology Institute, Morinaga Milk Industry Co. Ltd., 5-1-83 Higashihara, Zama, Kanagawa, Japan. t-odamak@morinagamilk.co.jp
authors.forename: T
authors.initials: T
authors.lastname: Odamaki
authors.forename: S
authors.initials: S
authors.lastname: Yonezawa
authors.forename: M
authors.initials: M
authors.lastname: Kitahara
authors.forename: Y
authors.initials: Y
authors.lastname: Sugahara
authors.forename: J-Z
authors.initials: JZ
authors.lastname: Xiao
authors.forename: T
authors.initials: T
authors.lastname: Yaeshima
authors.forename: K
authors.initials: K
authors.lastname: Iwatsuki
authors.forename: M
authors.initials: M
authors.lastname: Ohkuma
mesh_terms: D001431:Bacteriological Techniques / Q000379:methods*; D017931:DNA Primers / Q000235:genetics*; D017045:Lactococcus / Q000145:classification / Q000235:genetics*; D013294:Lactococcus lactis / Q000145:classification / Q000235:genetics; D016133:Polymerase Chain Reaction; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013045:Species Specificity
chemical_list: D017931:DNA Primers; D012336:RNA, Ribosomal, 16S