益生菌Lactobacillus gasseri real-time PCR快速鑑定方法   全文下載

Rapid Identification of Lactobacillus gasseri using real-time PCR

Lactobacillus gasseri（L. gasseri）已發現其能抑制致病菌及食物腐敗、具抗發炎特性、調節腸道環境等功能，常見於市售保健食品。然而許多文獻亦指出益生菌產品常有標示不清或成分不符的狀況，傳統生化檢驗方法耗時且有精準度不足之缺點，故為提升檢驗效率及精準度，本研究應用即時聚合酶鏈反應（real-time PCR）技術，建立L. gasseri之專一性快速鑑定方法。以ITS-23S rRNA基因設計引子探針組，並以91株參考菌株進行測試，結果顯示設計之引子及探針僅對目標參考菌株有專一性反應，且菌量低至約10 CFU/mL仍可偵測到螢光訊號，本研究開發之L. gasseri real-time PCR方法可提供L. gasseri之快速鑑定。

Lactobacillus gasseri is known for its ability to inhibit pathogenic bacteria and food spoilage, as well as for its anti-inflammatory properties and role in regulating intestinal environments. This bacterium is commonly present in retailed dietary supplements. However, numerous studies have highlighted the lack of clarity in labeling and discrepancies in the ingredients of these products. Traditional biochemical test methods are both time-consuming and imprecise. Therefore, in order to improve the efficiency and accuracy, this study aims at developing a rapid identification method for L. gasseri using real-time PCR. Primer-probe sets that target the ITS-23S rRNA genetic region of L. gasseri were designed and evaluated with 91 reference strains. The results demonstrated that the designed primers and probes were specific to the target reference strains of L. gasseri, and fluorescent signals could be detected even at bacterial concentrations as low as approximately 10 CFU/mL. The data showed the capability of this method a rapid and accurate technique for L. gasseri detection.