التغاير الجيني بين العزلات السريريه لبكتريا الاشريشيا القولونية == Genetic Diversity among Clinical Isolates of Escherichia coli
Author name:
سارة عادل عبيد حسوني السعدي
Supervisor name:
محمد صبري عبد الرزاق السعيد
General topic:
Medicine
Specific topic:
Microbiology
Degree:
Doctorate
University:
University of Babylon - Faculty Of Medicine
Language:
English
University location:
Babylon
First pages:
19T1726 - p.pdf
Abstract:
The present study includes 100 samples which were collected from patients suffering from urinary tract infections, vaginitis, diarrhea and wounds from both sexes and different ages, who attended to three main hospitals in Hilla city /Babylon governorate : Al - Hilla General Teaching Hospital, Babylon Hospital for Maternity and Pediatric and Mirgan Teaching Hospital during a period ranging from (February to June 2017).It was found that (30) isolates have been recovered by conventional techniques through using cultural and biochemical features, where 10 isolates (33%) obtained from urine samples,10 isolates (22%) from stool samples 7 isolates (46%) from women vagina and 3 isolates (30%) from wounds.In addition, the confirmatory identification of E. coli isolates were assessed by using Vitek2 compact system. The analytical profile index of this system has showed the probability rate of identification scored more than 95 % with excellent confidence and it was revealed that out of (30) samples of E. coli clinical isolates, only 27(90%) were confirmed as E. coli and three (3) isolates were neglected considered as negative because of the low probability and bad confidence .The internally transcribed spacer 16S - 23S rRNA regions of E.coli were analyzed with respect of 30 specimens of E.coli and it was revealed that out of (30) E. coli isolates, only (27) isolated were correctly identified as E. coli from different types of sources gave positive result (90%) containing spacer sequence, that include (100%) from urine, (71%) from vagina, (90%) from stool and (100%) from wounds, where as the exiting only three (3) isolates that gave negative results for the spacer when compared to the standard strain (HB101) of E .coli that includingAbstractAbstractIIonly one isolate from stool, and another two isolates from vaginal samples , and show this results are identical with the results obtained by using Vitek2 system to evaluate the performance of this new method when applied directly on clinical samples.Indeed, the study implicated the detection of genetic genetic genetic genetic genetic genetic diversity diversitydiversitydiversitydiversitydiversitydiversity among bacterial isolates was among bacterial isolates was among bacterial isolates was among bacterial isolates was among bacterial isolates was among bacterial isolates was among bacterial isolates was among bacterial isolates was among bacterial isolates was among bacterial isolates was among bacterial isolates was among bacterial isolates was among bacterial isolates was among bacterial isolates was among bacterial isolates was among bacterial isolates was among bacterial isolates was among bacterial isolates was among bacterial isolates was among bacterial isolates was among bacterial isolates was performed performedperformedperformedperformedperformedperformedperformed through using (ERIC - PCR) , (REP - PCR) and (BOX - PCR) and they were confirmed present in E. coli.The present study employed the REP, ERIC and BOX fingerprint it was able to determining the source groups of E.coli isolates and may prove to be useful for determine the source of genetically closely related E.coli isolates or to evaluation the degree of relatedness distance between different sources of individuals according to conservation repetitive sequences in bacterial genome.The scoring data from UPGMA found that the E.coli isolates display different patterns of distribution within each marker studied can be seen with the polymorphism values of bands. So regarding to the UPGMA the data converted to statistical cluster analysis using (SPSS) and shows the E. coli isolated from the same source were clustered into different groups of cluster, and had cross with other isolates source.Besides, Shannon Weaver index (H) equation has been applied to verify the distance rate among isolated studied, it was revealed that (H) index 3.11, 3.15 , 3.07 for REP, ERIC and BOXA of E.coli fingerprinting molecular marker respectively.Although , all 27 clinical samples of E.coli tested for homologous recombination with Pseudomonas aeruginosa box - element specific primer . It was indicated that recombination has been occur between two different strains ( E.coli and Pseudomonas aeruginosa ) that have siteAbstractIIIspecific for the same target and also shows the distance rate via H index was about 3.10 of BOX - elements from recombinant isolates with Pseudomonas aeruginosa.The molecular detection study of CRISPR 2.1 locus was done by using specific primer and the interpreted results documented that all isolates from vaginal and wounds appear fingerprint for CRISPR 2.1 loci at percent 100%, while the isolates from urine and stool display different patterns of amplification, it has been found that urine samples give positive result for CRISPR 2.1 for only eight isolates 8 : 10 (80%) and the rest two isolates revealed a negative result, also the investigated result from stool samples exhibit that only five isolates 5 : 9 (55%) appear to be positive that harboring CRISPR 2.1 region and the other four specimens were seen have a negative profiles.Moreover, all the identified isolates of E. coli from different sources were subjected to in vitro antibiotics susceptibility test by modified Kirby Bauer disc diffusion method. Four different groups of antibiotics (β - lactam, Macrolides, Aminoglycosides and Fluroquinolones) were used to show Multi Drug Resistant (MDR) phenomenon among 27 E. coli isolates groups.Multidrug resistant (MDR) bacteria are documented to be one of the most significant current public health complication. Since it has been found that E.coli isolates have highest rate of resistance at (70.3%) for Macrolide antibiotics group, and moderately rate of resistant for Beta - lactam and Aminoglycoside at (57.7%) and (55.5%) respectively. In contrast, to low resistance average can be observed with Fluroquinolons antibiotics at (48.1%). Also the result revealed that most E. coli isolates are sensitive at rate (14.8% and 18.5%) to Aztreonam and Meropenem respectively. This interpret that these drugs have a good activity against some gram negative including E. coli.
