Designing Two Synthetic Constructs for Real Time PCR Detection of Francisellatularensis and Ebola Virus

Background: generally, timely diagnosis of micro-organisms is very important to preventmany diseases. many methods can detect micro-organisms like culture-basedmethods and molecular methods. the molecular methods are usually preferred becausethey provide fast and reliable results. in some cases, microbial strains are not accessible,and there is no safety to work with them; therefore, synthetic constructs whichare designed according to the available sequences in databases can be used as a positivecontrol for detection of them.methods: in this study, a synthetic construct was designed for molecular detection offrancisella tularensis (f. tularensis) and the ebola virus by multiplex real-time pcrreaction. for this, sequences were taken from databases and then multiple alignmentswere done by software. also, conventional pcr and two models of real-timepcr (sybr green and taqman) were applied. finally, multiplex real-time pcr wasperformed.results: the synthetic construct was designed and used for conventional pcr andmultiplex pcr. the results of common pcr showed a single band at 148 bp and 167bp in 1.5% agarose gel stained by ethidium bromide for f. tularensis and ebola virus,respectively. also, a dual-band at 148 and 167 bp was observed in multiplex pcr. resultsof real-time pcr showed a limit of detection about 0.1 pg of plasmid/μl.conclusion: in conclusion, the designed construct can be used as a positive control foran accurate diagnosis of these micro-organisms without any biological danger for laboratorystaff. so, this method is useful for diagnosis of these agents in food, water,and blood samples.