The main purpose of this project is to improve the previous project which adopted quantum dots and microbeads to generate new technology for detection environmental microorganisms. The new technology aims to measure the quantity and quality by simple labeled methods thus the detection procedure will be short and faster than current methods, such as DNA chips. We proposed the quantum dot probes combined with microbead linked specific oligonucleotides following finger printing patterns as detection system that will be able to provide higher sensitivity and specificity with qualitative detection and quantitative determination. For detection of environmental microbes is the important issue to trace the pollution of environment. The tranditional methods required couple days to culture and verify, however, many microbes cannot be culture. Our novel technology ‘Quantum Dots-Finter Pringing Detection System’ is able to detect directly from water sample without culture or gene amplification. We selected 10 environmental bacterial spp and 4 adenovirus types as our research targets. Currently, we have followed the schedule of project to achieve : 1) microbead- Poly-methyl methacrylate (PMMA), 22~24 m, was linked with specific oligonucleotides from 10 environmental bacterial genuses, 2) quantum dots were linked to oligonucleotides (inrernal control and reporter for 10 environmental bacterial genuses individually) probes by click reaction, 3) The detection limit, qualitative detection and quantitative determination of microbed-quantum dot system were tested for each bacterial genuses, 4) the system of microbed-quantum dot-RFLP was tested and their standard protocols were made, 5) the detection limit of microbed-quantum dot-RFLP system for each bacterial genus was confirmed, all of those 10 environmental bacterial genuses can reach 1ng of detection limit to our system, 6) the linear regression of microbed-quantum dot-RFLP system for each bacterial genus was confirmed, all of those 10 environmental bacterial genuses’s linear regression over r2=0.9, 7) the sensitivity and specificity of microbed-quantum dot-RFLP system for each bacterial genus was confirmed, no false positive and false negative occured . These data have matched the requirement of the EPA proposal.

Finger printing;Quantum dots;Single oligonucleotide specific probes