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Novel multiplex polymerase chain reaction primer set for identification of Lactococcus species
Author(s) -
Odamaki T.,
Yonezawa S.,
Kitahara M.,
Sugahara Y.,
Xiao J.Z.,
Yaeshima T.,
Iwatsuki K.,
Ohkuma M.
Publication year - 2011
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1111/j.1472-765x.2011.03028.x
Subject(s) - lactococcus lactis , lactococcus , biology , multiplex polymerase chain reaction , polymerase chain reaction , 16s ribosomal rna , primer (cosmetics) , microbiology and biotechnology , ribosomal rna , bacteria , genetics , gene , chemistry , lactic acid , organic chemistry
Aims: 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. Methods and 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. Significance and Impact of the Study: This work provides an efficient method for identification of lactococcal strains of industrial importance.