Antibiotic Resistance Detection in Microbial Pathogens

The incidence of infectious sexually transmitted diseases has been increasing almost annually on a global scale including Neisseria gonorrhoeae. In addition, there is an increased antimicrobial resistance in microbial pathogens such as N. gonorrhoeae to different antibiotic drugs, i.e. the Ciprofloxacin resistant strains of N. gonorrhoeae have been reported in numerous developed countries.
The aims of this project is to genotypically characterize antibiotic resistance of microbial pathogens by short DNA-sequencing. As a model system, a known group of N. gonorrhoeae has been used for this study. The antibiotic resistance has been related to mutations in gyrA gene in the DNA-gyras protein coding region. We have investigated gyrA for drug resistance by conventional methods, which are time-consuming, subjective and labor-intensive. By applying Pyrosequencing technology for short DNA-sequencing, we have been able to genotype these mutations and associate them to antimicrobial drug resistance. The advantages of using Pyrosequencing technology over conventional gel-based sequencing methods are rapid real-time sequence readouts as well sequencing from the position of interest directly downstream of the sequencing primer.