Sodium Deoxycholate-Propidium Monoazide Droplet Digital PCR for Rapid and Quantitative Detection of Viable...

As a famous probiotic, Lacticaseibacillus rhamnosus HN001 is widely added to probiotic products. Different L. rhamnosus strains have different probiotic effects, and the active HN001 strain is the key to exerting probiotic effects, so it is of great practical significance for realising the detection of L. rhamnosus HN001 at the strain level in probiotic products. In this study, strain-specific primer pairs and probes were designed. A combined treatment of sodium deoxycholate (SD) and propidium monoazide (PMA) inhibited the amplification of dead bacterial DNA, establishing a SD-PMA-ddPCR system and conditions for detecting live L. rhamnosus HN001 in probiotic powders. Specificity was confirmed using type strains and commercial strains. Sensitivity tests with spiked samples showed a detection limit of 10⁵ CFU/g and a linear quantification range of 1.42 × 10⁵-1.42 × 10⁹ CFU/g. Actual sample testing demonstrated the method's efficiency in quantifying HN001 in compound probiotic products. This method offers a reliable tool for the rapid and precise quantification of viable L. rhamnosus HN001, crucial for the quality monitoring of probiotic products.