Open AccessRapid identification of 11 human intestinal Lactobacillus species by multiplex PCR assays using group‐ and species‐specific primers derived from the 16S–23S rRNA intergenic spacer region and its flanking 23S rRNA
Author(s) -
Song YuLi,
Kato Naoki,
Liu ChengXu,
Matsumiya Yoshiko,
Kato Haru,
Watanabe Kunitomo
Publication year - 2000
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.2000.tb09155.x
Subject(s) - lactobacillus salivarius , biology , lactobacillus gasseri , lactobacillus , lactobacillus fermentum , microbiology and biotechnology , lactobacillus crispatus , lactobacillus paracasei , 23s ribosomal rna , lactobacillus reuteri , multiplex polymerase chain reaction , lactobacillus rhamnosus , polymerase chain reaction , lactobacillus plantarum , bacteria , genetics , gene , ribosome , lactic acid , rna
Rapid and reliable two‐step multiplex polymerase chain reaction (PCR) assays were established to identify human intestinal lactobacilli; a multiplex PCR was used for grouping of lactobacilli with a mixture of group‐specific primers followed by four multiplex PCR assays with four sorts of species‐specific primer mixtures for identification at the species level. Primers used were designed from nucleotide sequences of the 16S–23S rRNA intergenic spacer region and its flanking 23S rRNA gene of members of the genus Lactobacillus which are commonly isolated from human stool specimens: Lactobacillus acidophilus , Lactobacillus crispatus , Lactobacillus delbrueckii (ssp. bulgaricus and ssp. lactis ), Lactobacillus fermentum , Lactobacillus gasseri , Lactobacillus jensenii , Lactobacillus paracasei (ssp. paracasei and ssp. tolerans ), Lactobacillus plantarum , Lactobacillus reuteri , Lactobacillus rhamnosus and Lactobacillus salivarius (ssp. salicinius and ssp. salivarius ). The established two‐step multiplex PCR assays were applied to the identification of 84 Lactobacillus strains isolated from human stool specimens and the PCR results were consistent with the results from the DNA–DNA hybridization assay. These results suggest that the multiplex PCR system established in this study is a simple, rapid and reliable method for the identification of common Lactobacillus isolates from human stool samples.