Degenerate Primer Based PCR for Rapid and Accurate Detection of Infectious Pathogens, Acinetobacter baumannii, and Staphylococcus aureus, with the Targeting of FtsZ Gene

Abstract

Aims: FtsZ is a highly conserved protein that is found in most of the major groups of bacteria. Considering the importance of the FtsZ gene in bacteria, in this study, Acinetobacter baumannii, Escherichia coli, and Staphylococcus aureus pathogens were detected by a new degenerate primer using PCR method with the targeting of FtsZ gene. Methods: Of each bacterium 30 clinical samples were collected from clinical diagnostic laboratories. For the specificity, The FtsZ gene was amplified using uniplex and triplex PCR with a degenerate primer as forward and three specific reverse primers for A. baumannii, E. coli, and S. aureus samples and some other species as negative control samples. The sensitivity of the primers was determined by PCR using a serial dilution of the genomic DNA template (100 ng to 1 pg). Results: The results showed that the PCR assay is positive for all of the A. baumannii, E. coli, and S. aureus samples (100%) while the PCR amplification was negative for other species. Conclusions: According to the results, the developed PCR assay using FtsZ gene is a reliable, fast, easy to perform and high specific and sensitive diagnostic method for early detection of the A. baumannii, E. coli, and S. aureus.