文献类型：期刊文献

英文题名：Development of PMA-qPCR assay to accurately and reproducible quantify viable bacteria of Paenibacillus polymyxa

作者：Guo, Jiacai[1];Fan, Fei[1];Wang, Weiliang[1];Wan, Minxi[1,2];Li, Yuanguang[1,2]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China;[2]Mailbox 301, Meilong Rd 130, Shanghai 200237, Peoples R China

年份：2023

卷号：76

期号：11

外文期刊名：LETTERS IN APPLIED MICROBIOLOGY

收录：;EI(收录号：20242116106753);WOS:【SCI-EXPANDED(收录号:WOS:001107319500001)】；

基金：This research was supported by the National Key Research and Development Program of China (2017YFD0201107-2-3).

语种：英文

外文关键词：Paenibacillus polymyxa; propidium monoazide; quantitative PCR; viable bacteria number; plate count method

摘要：Paenibacillus polymyxa is an important biocontrol bacterium. The combination of propidium monoazide (PMA) and quantitative polymerase chain reactionq (qPCR) has proven effective in quantifying live bacteria from various microorganisms. The objective was to create a PMA-qPCR assay to precisely and consistently measure the number of living bacteria of biocontrol P. polymyxa. The primers were designed for the spo0A gene of P. polymyxa HY96-2. The optimal conditions for treating the target strain with PMA were a PMA concentration of 15 mu g/mL, an incubation time of 5 min, and an exposure time of 10 min. The PMA-qPCR method had a limit of quantification (LOQ) of 1.0 x 103 CFU/mL for measuring the amount of viable P. polymyxa bacteria. The PMA-qPCR method is more sensitive than the qPCR method in detecting viable bacteria in the mixtures of viable and dead bacteria. The accuracy and reproducibility of quantifying viable P. polymyxa bacteria using the PMA-qPCR method were higher compared to the plate count method.