The comparison of MAMA PCR and SSCP PCR to study chromosomal resistance against Ciprofloxacin and Nalidixic acid in Escherichia coli and Klebsiella pneumoniae

Abstract

The mutation in gyrA and parC genes alters amino acids. Also, it causes resistance against Fluoroquinolones in E. coli and K. pneumoniae. The purpose of this study was to diagnose the significant mutation of gyrA (ser83-asp87) and parC (ser80-glu84) genes through using MAMA PCR and SSCP PCR methods. In so doing, the isolated samples were collected. Then, utilizing agar disc diffusion method, the researchers performed antibiotic sensitivity test. Moreover, Fluoroquinolones resistance was confirmed by E-test method (MIC experiment). Furthermore, the obtained data from MAMA PCR method were sequenced accidentally. According to the findings, among 103 isolated samples, 65 samples (63/2%) were belonged to E. coli and 38 samples (36/8%) to K. pneumoniae. In all E. coli that resisted to Ciprofloxacin, at least one mutation were observed. Also, at least one mutation was observed in all K. pneumoniae samples that resisted to Ciprofloxacin. However, four mutation points were detected for each of seven samples and, interestingly, there was no mutation in five sensitive samples to Ciprofloxacin. In addition, the results revealed that the mutation in gyrA and parC genes was closely related to Quinolones resistance. Based on the findings, preparing an infection control program in Iran is highly required.

Keywords: Antibiotic resistance; Escherichia coli; Klebsiella pneumoniae; MAMA PCR; Quinolones; SSCP PCR.