Real-time PCR for Detection of Lactobacillus rhamnosus Based on Dual Priming Oligonucleotide System

Objective To develop a method for the detection of Lactobacillus rhamnosus.Method The species-specific DPO (dual priming oligonucleotide) primers were designed based on thegyrB gene sequences of L.rhamnosus.A SYBR Green I real-time PCR assay based on DPO primers was developed for the detection of L.rhamnosus.The specificity and sensitivity of the assay have been estimated.Result The results showed that this method was of high specificity,and only two L.rhamnosus strains can be amplified.The other 10 L.spp.strains gave negative results.The detection sensitivity of this real-time PCR was more than 10 times higher than the conventional PCR.Conclusion SYBR Green I real-time PCR method established in this study can detect L.rharanosus accurately and rapidly in the microbial fertilizers and other probiotic products.