Design of Bioluminescence Sensor System for Rapid Detection of Bacteria in Samples without Cultivation

Abstract

Based on the principle of ATP bioluminescent reaction, we design a portable bioluminescence sensor system to detect bacteria in samples rapidly, which included an optical sensing cell, a flow injection unit and a center processing unit. A fast ATP extraction technique was developed by using trichloro acetic acid as extraction reagent Experimental results showed that the intracellular ATP could be released from bacterial cells sufficiently within 5 min by TCA In the presence of Lucifer ace the reaction between ATP and lucifer5in would generate light and the luminescence intensity RLU was proportional to the amount of bacteria existed in the sample. The dynamic range of the sensor system is 10-107 CFU /ml. The linear regression coefficient was 0.976, and the detection limit was 10CFU /ml. The whole testing time needed was about 10 min. Compared with the plate count method, bacteria concentration in samples could be detected rapidly by using the portable sensor system without cultivation.