Establishment and application of real-time fluorescence quantitative PCR for detection of Acetilactobacillus jinshanensis

Abstract:

[Objective] To establish a real-time quantitative method PCR (RT-qPCR) for the detection of Acetilactobacillus jinshanensis in the fermentation of Chinese traditional fermented vinegar and baijiu.[Methods] The specific gene in the Acetilactobacillus jinshanensis genome was screened for the design of PCR-sequence specific primer. The specificity and accuracy of specific primers were verified by PCR with isolated strains and vinegar Pei. The RT-qPCR method was established to analyze the content of Ac. jinshanensis in vinegar, jiupei and cupei from different regions.[Results] We designed a pair of primers with product size of 199 bp, GC content of 55% and T_m value of 59℃ with phenylalanine-tRNA ligase subunit α (encoding pheS gene) of Ac. jinshanensis. The RT-qPCR method has potent specificity, high sensitivity, good repeatability and versatility, and its detection range is 2.24-10.24 lg(copies/μL). The method can be applied for the detection of Ac. jinshanensis in the fermentation of cupei and jiupei from different regions.[Conclusion]] The RT-qPCR method can be used for the specific identification and accurate quantification of Ac. jinshanensis in the fermentation of vinegar and baijiu.

Key words:Zhenjiang aromatic vinegar;Acetilactobacillus jinshanensis;specific primers;RT-qPCR