caa a22 4500 605506612 CHVBK 20210128100628.0 cr unu---uuuuu 210128e20150801xx s 000 0 eng 10.1007/s00253-015-6715-x doi (NATIONALLICENCE)springer-10.1007/s00253-015-6715-x Quantifying viable Vibrio parahaemolyticus and Listeria monocytogenes simultaneously in raw shrimp [Elektronische Daten] [Zhaohuan Zhang, Haiquan Liu, Yang Lou, Lili Xiao, Chao Liao, Pradeep Malakar, Yingjie Pan, Yong Zhao] A novel TaqMan-based multiplex real-time PCR method combined with propidium monoazide (PMA) treatment was firstly developed for the simultaneous quantification of viable Vibrio parahaemolyticus and Listeria monocytogenes in raw shrimp. The optimization of PMA concentration showed that 100μM was considered optimal to effectively inhibit 108CFU/mL dead cells of both bacteria. The high specificity of this method was confirmed on tests using 96 target and non-target strains. The optimized assay could detect as low as 101-102CFU/g of each strain on the artificially contaminated shrimp, and its amplification efficiencies were up to 100 and 106% for V. parahaemolyticus and L. monocytogenes, respectively. Furthermore, this assay has been successfully applied to describe the behavior of these two pathogens in raw shrimps stored at 4°C. In conclusion, this PMA TaqMan-based multiplex real-time PCR technique, where the whole procedure takes less than 5h, provides an effective and rapid tool for monitoring contamination of viable V. parahaemolyticus and L. monocytogenes in seafood, improving seafood safety and protecting public health. Springer-Verlag Berlin Heidelberg, 2015 TaqMan-based multiplex real-time PCR nationallicence Propidium monoazide nationallicence Viable foodborne pathogens nationallicence Vibrio parahaemolyticus nationallicence Listeria monocytogenes nationallicence Raw shrimp nationallicence Zhang Zhaohuan College of Food Science and Technology, Shanghai Ocean University, 999# Hu Cheng Huan Road, 201306, Shanghai, China aut Liu Haiquan College of Food Science and Technology, Shanghai Ocean University, 999# Hu Cheng Huan Road, 201306, Shanghai, China aut Lou Yang College of Food Science and Technology, Shanghai Ocean University, 999# Hu Cheng Huan Road, 201306, Shanghai, China aut Xiao Lili College of Food Science and Technology, Shanghai Ocean University, 999# Hu Cheng Huan Road, 201306, Shanghai, China aut Liao Chao College of Food Science and Technology, Shanghai Ocean University, 999# Hu Cheng Huan Road, 201306, Shanghai, China aut Malakar Pradeep College of Food Science and Technology, Shanghai Ocean University, 999# Hu Cheng Huan Road, 201306, Shanghai, China aut Pan Yingjie College of Food Science and Technology, Shanghai Ocean University, 999# Hu Cheng Huan Road, 201306, Shanghai, China aut Zhao Yong College of Food Science and Technology, Shanghai Ocean University, 999# Hu Cheng Huan Road, 201306, Shanghai, China aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/15(2015-08-01), 6451-6462 0175-7598 99:15<6451 2015 99 253 https://doi.org/10.1007/s00253-015-6715-x text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6715-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Zhang Zhaohuan College of Food Science and Technology, Shanghai Ocean University, 999# Hu Cheng Huan Road, 201306, Shanghai, China aut NATIONALLICENCE 700 1- Liu Haiquan College of Food Science and Technology, Shanghai Ocean University, 999# Hu Cheng Huan Road, 201306, Shanghai, China aut NATIONALLICENCE 700 1- Lou Yang College of Food Science and Technology, Shanghai Ocean University, 999# Hu Cheng Huan Road, 201306, Shanghai, China aut NATIONALLICENCE 700 1- Xiao Lili College of Food Science and Technology, Shanghai Ocean University, 999# Hu Cheng Huan Road, 201306, Shanghai, China aut NATIONALLICENCE 700 1- Liao Chao College of Food Science and Technology, Shanghai Ocean University, 999# Hu Cheng Huan Road, 201306, Shanghai, China aut NATIONALLICENCE 700 1- Malakar Pradeep College of Food Science and Technology, Shanghai Ocean University, 999# Hu Cheng Huan Road, 201306, Shanghai, China aut NATIONALLICENCE 700 1- Pan Yingjie College of Food Science and Technology, Shanghai Ocean University, 999# Hu Cheng Huan Road, 201306, Shanghai, China aut NATIONALLICENCE 700 1- Zhao Yong College of Food Science and Technology, Shanghai Ocean University, 999# Hu Cheng Huan Road, 201306, Shanghai, China aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/15(2015-08-01), 6451-6462 0175-7598 99:15<6451 2015 99 253