1. Malek ashtar university of technology 2. Malek ashtar university of technology 3. Malek ashtar university of technology 4. Malek ashtar university of technology
Abstract
Introduction
Q fever is a zoonotic disease that caused by coxiella burnetii, a gram-negative and intracellular bacterium. today, traditional diagnosis of q fever in human and animals are based on serological methods that is time consuming and laborious. on the other hand, for detection of c.burnetii in the environment, in order to investigate possible routes of dissemination and transmission, serology method, cannot be applied. therefore, molecular detection is preferred because this assay is fast and reliable in most conditions.
Methods
This bacterium is belonging to level 3 biosafety, so we designed a synthetic cassette which, consist of three specific regions according to specific genes for detection of c.burnetti. this cassette including com1, icd & is1111 genes to develop a multiplex pcr assay to identification of q fever agent. this cassette can be used as a positive control for multiplex pcr assay.
Results
According to designing of synthetic cassette, data showed that 3 specific bands at 300bp, 140bp & 90bp could be observed in 2% agarose gel electrophoresis. development of this synthetic cassette as a positive control and multiplex pcr assay may be useful for identification of infected products.
Conclusion
Synthetic cassettes are useful tools for detection of microbial agents that, are need biosafety level 3 or more conditions. we can use these tools to prevent of transmission to laboratory personals.
