Application of propidium monoazide coupled with quantitative PCR to evaluate cell viability of Bifidobacterium animalis subsp. lactis in a non-dairy probiotic beverage

Abstract

Purpose: In this study, a PMA-qPCR assay was developed for the enumeration of Bifidobacterium animalis subsp. lactis BB-12 viable cells in a non-dairy probiotic beverage. Methods: Probiotic viability was monitored in three formulations of probiotic passion fruit juice microencapsulated by spray drying, during 30 days of storage at 4 °C. Viable cells were quantified using qPCR and PMA-qPCR assays targeting tuf gene and by plate counting method. Results: The limit of detection for all samples was 103 genome copies, corresponding to 21.3 pg of DNA. Higher CFU values were obtained for B. lactis BB-12 enumeration by qPCR, when compared to those obtained by PMA-qPCR and plate count, for all probiotic juice microcapsules. Similar quantification values were obtained by PMA-qPCR and plate counting for all samples and remained above 8 log CFU/g during the storage period. Conclusion: These results demonstrated that the PMA-qPCR technique is a promising approach for B. lactis BB-12 viable cell enumeration in complex matrices such as passion fruit juice microcapsules. This PMA-qPCR assay allowed the achievement of reliable results faster than with the traditional plate counting method.