PMA-qPCR method for accurately quantifying live bacteria of animal bifidobacterium subsp. Lactis for food

Through PMA-qPCR technology combined with specific primers, the accurate identification and quantitative detection of animal Bifidobacterium milk subspecies in complex probiotic products was achieved, and the problem of long inspection cycles of traditional methods and the inability to synchronize the number of live bacteria/strains was solved, improving the sensitivity and accuracy of the detection.

CN119710051APending Publication Date: 2025-03-28CHINA NAT RES INST OF FOOD & FERMENTATION IND CO LTD
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Patent Information

Application Number
CN202510083240.0
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2025-01-20
Publication Date
2025-03-28

AI Technical Summary

Technical Problem

It is difficult for the prior art to achieve accurate identification and quantitative detection of animal Bifidobacterium milk subspecies in complex probiotic products, especially in complex probiotic systems. Traditional methods have a long test cycle and cannot synchronously detect the number of live bacteria in each strain/strain.

Method used

PMA-qPCR technology combined with specific primers to distinguish live dead bacteria through PMA dye, and real-time fluorescence quantitative PCR was used to accurately identify and quantitatively detect live bacteria in animal Bifidobacterium milk subspecies.

Benefits of technology

It has achieved efficient and accurate identification and quantitative detection of animal Bifidobacterium milk subspecies in complex probiotic products, improved detection sensitivity and accuracy, and met the high requirements for detection of bacterial species activity and quantity in complex probiotic systems.

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Abstract

The invention discloses a method for rapidly detecting animal bifidobacterium subsp. Lactis, and particularly relates to a group of specific primer groups for detecting the animal bifidobacterium subsp. Lactis, a qPCR (quantitative polymerase chain reaction) viable bacterium detection method combined with PMA (polymethyl acrylate) dye and application of the qPCR viable bacterium detection method. The invention provides application of a nucleotide sequence (SEQ ID NO: 1) in identification of bifidobacterium animalis subsp. Lactis strains. Specific PCR primers are shown as SEQ ID NO: 2 and SEQ ID NO: 3. The qPCR analysis is carried out by combining PMA dye with the primer group, so that the rapid and accurate quantitative detection of the animal bifidobacterium subsp. Lactis viable bacteria is realized. The method has the remarkable advantages of being high in detection sensitivity, high in specificity and short in consumed time, targeted viable bacteria quantitative detection can be conducted on the bifidobacterium animalis subsp. Lactis in the compound probiotic product, reliable technical support is provided for the quality control and detection department of the probiotic product, and the important industrial application value is achieved.
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