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مقارنة تشخيصية جزيئية وبكتريولوجسة لبكتريا الزائفة الزنجارية المعزولة من مرضى السكري النوع الثاني وامراض اخرى == Comparison the Molecular and Conventional Identification of Pseudomonas aeruginosa isolated from Diabetes Type II and other Diseases
Author name:
سناء محمد حبيب الشمري
Supervisor name:
اسماء محمد صالح المهيدي
General topic:
Biology
Specific topic:
Microbiology - Immunology
Degree:
Master
University:
University of Baghdad
Language:
English
University location:
Baghdad
First pages:
24T3335 - p.pdf
Abstract:
Fifty clinical samples suspected as Pseudomonas aeruginosa were collected from patients with different diseases, diabetic foot ulcers, U.T.I., wounds, burns, keratitis, external otitis from September 2015, till February 2016, from hospitalized patients in four different Baghdad hospitals (Al - Kindi General Teaching Hospital, Al - Yarmuk General Teaching Hospital, Alkadhmiya General Teaching Hospital, and AL - Shiakh Zaied Hospital).The study population targeted both gender from different ages groups.The collected samples were cultured on different selective and differential media and tested biochemically in order to identify P.aeruginosa, then the identification was ensured by using two molecular methods which are conventional PCR in the detection of ecfx gene and 16SrDNA gene and real time PCR with commercial kit from sacace (qualitative detection of P.aeruginosa) and with gyrB gene using specific primers to each gene.Sensitivity for each test was measured then compared with other tests to determine the test efficiency. The results showed that the sensitivity for API20NE test was (80%), the sensitivity for molecular tests as flows : ecfx test was (92%), 16SrDNA test was (96%), the commercial kit test was (98%) and for gyrB test was (92%).The present study shows that using more than one target gene is reducing the probability of misidentification of P.aeruginosa that may be caused by variation in primer or probe targets.According to the type of infection the comparison was done. The results of P.aeruginosa detection for diabetic foot isolates were compared with other isolates. The results showed that there is a remarkabledifference in the API20NE test; as well as, with the oxidase test (according to the time that oxidase test need it to give the dark purple color as a positive result), for the mucoid strains of this bacterium. The results of detection isolates by using gyrB gene were differed too.Then a comparison in simplicity, cost, accuracy and time of P.aeruginosa identification between molecular methods and API20NE test (biochemical test) and between molecular tests themselves were performed. The results of these comparisons show that the molecular methods are better than API test in all aspects of the comparison. Real time PCR is better than conventional PCR in simplicity, accuracy and time but not in cost.All isolates were tested for antibiotic sensitivity using nine antibiotics. The results showed that most isolates were sensitive to Levofloxacin (94%), Ticarcillin - clavulanic acid (Timinim) (88%), Gentamicin (82%) and (80%) to both Pipracillin and Imipenem, while the isolates appeared resistant to another antimicrobials represented by (72%) to Augmentin, (34%) to Ceftazidime, (38%) to Colistin, and (28%) to Ciprofloxacin. In the present study, Levofloxacin represents the most effective antibiotic against P. aeruginosa isolates with the all types of infection.The present study improves the importance of the rapid and accurate detection of the bacterial types and thus giving the right antibiotics to patients to obtain rapid recovery and prevent any complications that may lead to death.
