Development of a Direct and Rapid Detection Method for Viable but Non-culturable State of Pediococcus acidilactici.

Yu Guan,Yanrui Ye,Ling Chen,Yang Zeng,Kan Wang,Tengyi Huang

doi:10.3389/fmicb.2021.687691

Abstract

Pediococcus acidilactici may significantly reduce the pH-value, and thus has different influence, including serving as a probiotic in human microbiota but a spoilage in human food as it could change the flavor. Pediococcus acidilactici is also capable of entering into the viable but non-culturable (VBNC) state causing false negative results of standard culture-based detection method. Thus, development of detection method for VBNC state P. acidilactici is of great significance. In this study, propidium monoazide (PMA) combined with cross priming amplification (CPA) was developed to detect the VBNC cells of P. acidilactici and applied on the detection in different systems. With detection limit of 104 cells/ml, high sensitivity, and 100% specificity, PMA-CPA can successfully detect VBNC cells of P. acidilactici and be applied in with high robustness.

Highlights

Pediococcus acidilactici may significantly reduce the pH-value, and has different influence, including serving as a probiotic in human microbiota but a spoilage in human food
In this study, first we developed a rapid, sensitive, and specific detection assay on P. acidilactici based on cross priming amplification (CPA) methodology
The results showed that the viable but non-culturable (VBNC) cells can be detected successfully by propidium monoazide (PMA)-CPA (Figure 4A)

Summary

Introduction

Pediococcus acidilactici may significantly reduce the pH-value, and has different influence, including serving as a probiotic in human microbiota but a spoilage in human food. P. acidilactici has been considered to be a probiotic bacteria for human beings, and a typical type of lactic acid bacteria (LAB), which have been well-documented to be widely existing in human microbiota. Such probiotic bacteria have been well-studied to significantly aid in the microbiota balance, and once altered, various types of human diseases could occur. Probiotic LAB is an important type of human probiotic and significantly aids in the prevention of urinary tract infection. A major concern still remains, as a large proportion of such

Methods

Results

Conclusion