title: Simultaneous discrimination of species and strains in Lactobacillus rhamnosus using species-specific PCR combined with multiplex mini-sequencing technology.
abstract: This study described the use of species-specific PCR in combination with SNaPshot mini-sequencing to achieve species identification and strain differentiation in Lactobacillus rhamnosus. To develop species-specific PCR and strain subtyping primers, the dnaJ gene was used as a target, and its corresponding sequences were analyzed both in Lb. rhamnosus and in a subset of its phylogenetically closest species. The results indicated that the species-specific primer pair was indeed specific for Lb. rhamnosus, and the mini-sequencing assay was able to unambiguously distinguish Lb. rhamnosus strains into different haplotypes. In conclusion, we have successfully developed a rapid, accurate and cost-effective assay for inter- and intraspecies discrimination of Lb. rhamnosus, which can be applied to achieve efficient quality control of probiotic products.
authors.affiliation: Bioresource Collection and Research Center, Food Industry Research and Development Institute, 331 Shih-Pin Road, Hsinchu 30062, Taiwan.
authors.forename: Chien-Hsun
authors.initials: CH
authors.lastname: Huang
authors.affiliation: National Health Research Institutes, Institute of Biotechnology and Pharmaceutical Research, 35 Keyan Road, Zhunan, Miaoli County 35053, Taiwan.
authors.forename: Mu-Tzu
authors.initials: MT
authors.lastname: Chang
authors.affiliation: Bioresource Collection and Research Center, Food Industry Research and Development Institute, 331 Shih-Pin Road, Hsinchu 30062, Taiwan.
authors.forename: Lina
authors.initials: L
authors.lastname: Huang
authors.affiliation: Bioresource Collection and Research Center, Food Industry Research and Development Institute, 331 Shih-Pin Road, Hsinchu 30062, Taiwan. Electronic address: cws@firdi.org.tw.
authors.forename: Wen-Shen
authors.initials: WS
authors.lastname: Chu
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics; D015373:Bacterial Typing Techniques / Q000191:economics / Q000379:methods*; D017931:DNA Primers / Q000235:genetics; D052201:Lacticaseibacillus rhamnosus / Q000145:classification* / Q000235:genetics; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000191:economics / Q000379:methods; D017422:Sequence Analysis, DNA / Q000191:economics / Q000379:methods*; D013045:Species Specificity
chemical_list: D001426:Bacterial Proteins; D017931:DNA Primers
keywords: Lactobacillus rhamnosus; Species identification; Strain subtyping; dnaJ sequence