Application of the VPp1 bacteriophage combined with a coupled enzyme system in the rapid detection of Vibrio parahaemolyticus.
For rapid and quantitative detection of Vibrio parahaemolyticus, a method combining the specific lysis of bacteriophages with a bacterial luciferase-flavin mononucleotide:nicotinamide adenine dinucleotide oxidoreductase bioluminescent system in vitro was developed. A V. parahaemolyticus detection system was established by optimizing three main influencing factors: bacteriophage titer, volume ratio of the bacteriophage to its host bacterium, and lysis time. A standard curve between the number of bacteria and the luminescence intensity of the coupled enzyme system was studied and revealed a good linear relationship. More than 10(7)colony-forming units (cfu)·ml(-1) bacteria in pure culture and >10(8) cfu·ml(-1) bacteria in oyster samples were readily detected without pre-enrichment. Furthermore, >10(0) cfu·ml(-1) bacteria in oyster samples were readily detected after 4h of enrichment culture. Because of its rapid detection, high specificity, and simplicity in operation, this method is an effective tool for detecting living bacteria in food and environmental samples.
Food Safety Laboratory, College of Food Science and Engineering, Ocean University of China, Qingdao 266003, China.
Yong
Y
Peng
Food Safety Laboratory, College of Food Science and Engineering, Ocean University of China, Qingdao 266003, China.
Yanqiu
Y
Jin
Food Safety Laboratory, College of Food Science and Engineering, Ocean University of China, Qingdao 266003, China.
Hong
H
Lin
Food Safety Laboratory, College of Food Science and Engineering, Ocean University of China, Qingdao 266003, China. Electronic address: snow@ouc.edu.cn.
Jingxue
J
Wang
Food Safety Laboratory, College of Food Science and Engineering, Ocean University of China, Qingdao 266003, China; Microbiological Analytical Centre, FMRRC, PCSIR Labs. Complex Karachi, 75280, Pakistan.
Muhammad Naseem
MN
Khan
D000818:Animals; D001435:Bacteriophages / Q000378:metabolism*; D005486:Flavin Mononucleotide / Q000378:metabolism; D005516:Food Microbiology / Q000379:methods*; D008156:Luciferases / Q000378:metabolism; D008163:Luminescent Measurements / Q000379:methods*; D010125:Ostreidae / Q000382:microbiology; D012680:Sensitivity and Specificity; D013234:Stem Cells / Q000382:microbiology; D014735:Vibrio Infections / Q000175:diagnosis* / Q000382:microbiology; D014736:Vibrio parahaemolyticus / Q000378:metabolism*
D005486:Flavin Mononucleotide; D008156:Luciferases
Bacterial luciferase; Bacteriophage; FMN:NADH oxidoreductase; Rapid detection; Vibrio parahaemolyticus