Application of the VPp1 bacteriophage combined with a coupled enzyme system in the rapid detection of Vibrio parahaemolyticus

作者：Peng Yong; Jin Yanqiu; Lin Hong; Wang Jingxue^*; Khan Muhammad Naseem

来源：Journal of Microbiological Methods, 2014, 98: 99-104.

DOI：10.1016/j.mimet.2014.01.005

• Bacteriophage
• Vibrio parahaemolyticus
• Bacterial luciferase
• FMN:NADH oxidoreductase
• Rapid detection

For rapid and quantitative detection of Vibrio parahaemolyticus, a method combining the specific lysis of bacterio-phages 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 4 h 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.

• 出版日期2014-3
• 单位青岛大学; 中国海洋大学