Randa Hassan posteradmin
CHARACTERIZATION OF CIPROFLOXACIN RESISTANT PROTEUS MIRABILIS AND
EVALUATION THE EFFECTS OF GYRA AND GYRB, PARC AND PARE MUTATIONS IN
QUINOLONES RESISTANT URINARY ISOLATES
Randa H abdelkreem1, Leila M Abdelgadeir 1*, Miskelyemen A. Elmekki2*, Hisham N. Altayb3 and Mogahid M Elhassan 2
1Department of Microbiology, College of Medical Laboratory Science, Shendi University, Shendi, Sudan.
2Department of Clinical Laboratory Sciences, College of Applied Medical Sciences, Taibah University, KSA.
3 College of Medical Laboratory Science, Sudan University of Science and Technology, Sudan.
Background: Urinary tract infections (UTI) are major health problems affecting millions of people each year. As an opportunistic pathogen, Proteus mirabilis causes urinary tract infections. Ciprofloxacin is a recommended drug for the treatment of UTIs, but a progressive increase in fluoroquinolone resistance has been seen in clinical isolates. This study was a qualitative study, aimed to highlight the importance of using conventional and molecular techniques in the detection of different mutations within the genome of Proteus mirabilis isolates and their responsibility in fluoroquinolone .
Methods: In this study, a total of (120) Proteus mirabilis isolates from patients with symptoms of UTIs attending different hospitals in Khartoum State during the period from June 2016 to May 2017 were examined. Midstream urine samples were collected and cultured for UTI diagnosis and ciprofloxacin susceptibility. Then polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and sequencing was erformed for detection of GyrA, GyrB, ParC and ParE genes mutations of isolated P. mirabilis.
Results: The P. mirabilis showed (30%) resistant to ciprofloxacin. all samples mutate at (serine 83) of GyrA and (serine 84) of ParC by Hinf1 restriction endonuclease digestion. By sequencing, the mutations associated with ciprofloxacin resistant P. mirabilis in (33.3%) GyrA (Ser 83 to Ile) and (66.6%) ParC (Ser 81 to Ile). Also it revealed silent mutations at following codons of GyrB 474 leucine((100%), 585 valine (66.6%), 612 histidine(33.3%) and 639 asparagine(33.3%)) and ParE (469 isoleucine (66.6%), 531 aspartic (66.6%) and 533 glycine(33.3%)).
Conclusion: Direct Hinf1 digestion of PCR amplification is not suitable to screen serine (83) of GyrA and serine (84) of ParC mutations in P. mirabilis and the silent mutation in QRDR regions is not enough for ciprofloxacin resistance in Proteus mirabilis.