Novel multiplex polymerase chain reaction primer set for identification of Lactococcus species
Article first published online: 8 MAR 2011
© 2011 The Authors. Letters in Applied Microbiology © 2011 The Society for Applied Microbiology
Letters in Applied Microbiology
Volume 52, Issue 5, pages 491–496, May 2011
How to Cite
Odamaki, T., Yonezawa, S., Kitahara, M., Sugahara, Y., Xiao, J.-Z., Yaeshima, T., Iwatsuki, K. and Ohkuma, M. (2011), Novel multiplex polymerase chain reaction primer set for identification of Lactococcus species. Letters in Applied Microbiology, 52: 491–496. doi: 10.1111/j.1472-765X.2011.03028.x
- Issue published online: 7 APR 2011
- Article first published online: 8 MAR 2011
- Accepted manuscript online: 8 FEB 2011 03:14AM EST
- 2010/2069: received 16 November 2010, revised 19 January 2011 and accepted 2 February 2011
- L. chungangensis;
- L. fujiensis;
- L. garvieae;
- L. lactis;
- L. piscium;
- L. plantarum;
- L. raffinolactis;
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.