Rapid identification of probiotic Lactobacillus species by multiplex PCR using species-specific primers based on the region extending from 16S rRNA through 23S rRNA.
This study aimed to develop a novel multiplex polymerase chain reaction (PCR) primer set for the identification of seven probiotic Lactobacillus species such as Lactobacillus acidophilus, Lactobacillus delbrueckii, Lactobacillus casei, Lactobacillus gasseri, Lactobacillus plantarum, Lactobacillus reuteri and Lactobacillus rhamnosus. The primer set, comprising of seven specific and two conserved primers, was derived from the integrated sequences of 16S and 23S rRNA genes and their rRNA intergenic spacer region of each species. It was able to identify the seven target species with 93.6% accuracy, which exceeds that of the general biochemical methods. The phylogenetic analyses, using 16S rDNA sequences of the probiotic isolates, also provided further support that the results from the multiplex PCR assay were trustworthy. Taken together, we suggest that the multiplex primer set is an efficient tool for simple, rapid and reliable identification of seven Lactobacillus species.
ILDONG Research Laboratories, ILDONG Pharmaceutical Co., Ltd., 260-5, Eonnam-ri, Kuseong-eup, Yongin, Kyongki-do 449-915, Korea. hskwon@ildong.com
Hyuk-Sang
HS
Kwon
Eun-Hee
EH
Yang
Seung-Woo
SW
Yeon
Byoung-Hwa
BH
Kang
Tae-Yong
TY
Kim
D015373:Bacterial Typing Techniques / Q000379:methods*; D017931:DNA Primers; D021903:DNA, Ribosomal Spacer / Q000032:analysis* / Q000235:genetics; D007778:Lactobacillus / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction; D019936:Probiotics / Q000302:isolation & purification*; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012338:RNA, Ribosomal, 23S / Q000235:genetics
D017931:DNA Primers; D021903:DNA, Ribosomal Spacer; D012336:RNA, Ribosomal, 16S; D012338:RNA, Ribosomal, 23S