Molecular Detection of Proteus mirabilis isolated from Diabetic foot

Abstract

Diabetic foot ulcer (DFU) is one of the diabetic complications associated with major morbidity, mortality, and reduced quality of life and is the most serious complication of diabetes mellitus. A total of 118 wound swab samples were collected from diabetic foot ulcer patients attending five hospitals in Baghdad governorate (Al-Yarmouk Hospital, Al-sewaira Hospital, Medicine City/Baghdad Teaching Hospital, Endocrine gland center, AL-kadhimya Hospital) at a period of study from beginning  October 2019 to January 2020. Regarding to the age group factor,  the age group (50-59) years were more susceptible to the infection is constituting 49(49%) ,followed by the age group(40-49) years with 34(34%) then ,  the age group (60-69) years with  percentage at 15(15)%.and the age 70 years and above  with  percentage (2%)  Also,  the study indicated that the Diabetic foot ulcer was disrupted in male (58%) more than female (42 %).

All specimens were cultured on culture media including blood agar and MaCconkey agar. After the growth of bacteria, the isolates were identified by microscopic examination as well as the biochemical tests including the manual biochemical tests that include( oxidase , catalase, Simmon Citrate,  Motility , Indole , Urease , Methyl Red, Kligler iron agar (KIA) ,Lactose fermentation ,Voges-Proskauer) .The identification of P. mirabilis confirmed by using the systems API 20E and VITEK-2 system . A total of 18 isolates of P. mirabilis were identified. Other bacteria obtained were identified as Escherichia coli, Pseudomonas. aeruginosa, Klebsiella .pneumoniae and streptococcus spp.  in percentage recorded (37%) , (22%) (20.2%) and (11%) respectively. The genomic DNA of P. mirabilis isolates were extracted using wizard genomic DNA purification kit, the extracted genomic DNA was analyzed using 1% agarose gel electrophoresis, and then the concentration and purity of the extracted genomic DNA were determined using Nanodrop spectrophotometer device, to detect  P. mirabilis isolates by molecular methods, the extracted genomic DNA of these isolates was submitted for amplification to detect the specific gene 16S rRNA and aadA1 by the singleplex PCR assay.