Identification of trichophyton rubrum isolates by using traditional methods and RAPD-PCR

مقدم أطروحة جامعية

Salman, Zaynab Anas

مشرف أطروحة جامعية

al-Azami, Abd al-Karim Jasim Hashim
Karhut, Jasim M.

الجامعة

جامعة بغداد

الكلية

كلية العلوم

القسم الأكاديمي

قسم التقانة الإحيائية

دولة الجامعة

العراق

الدرجة العلمية

ماجستير

تاريخ الدرجة العلمية

2013

الملخص الإنجليزي

Fifty samples of skin scrapings, nail and hair from patients were obtained during the period beginning January/2011 till end April/2011 in Al-Yarmuk Hospital.

All specimens were subjected to direct KOH (10%) mount smear and culture on Sabourauds dextrose agar media to determine the dermatophyte species.

The direct KOH (10%) mount smear showed the presence of fungal elements in 40 (80%) of 50 human cases, while the growth of dermatophyte were positive in 40(80%) cases.

The main causative agents of human dermatophytes were as follow: Trichophyton rubrum 17(42.5%), Trichophyton mentagrophyte 13(32.5%), and Microsporum canis 10 (25%).

Identification of dermatophytes species depended on macroscopic colonial morphology and microscopically findings as well.

The PCR-based technique of randomly amplified polymorphic DNA (RAPD) was used to fingerprint for seventeen isolates of the fungus Trichophyton rubrum.

Genomic DNA of each isolate was extracted at final concentration of (750-1360) μg/ 1-2g of dry mycelium, and the purity of (1.3-1.6).

Each DNA sample was amplified with each 15 primers and the products were resolved electrophoretically on 1.2% agarose gel, stained with ethidium bromide and photographed under UV light.

II Two primers failed to support amplification but remaining 13 primers produced a total of (195) bands, of these bands 68%(126) bands were polymorphic.

The primers (A08 and GS04) were gave the highest number of polymorphic bands (15), while primers (E02 and A013) were gave the lowest number of polymorphic bands (6).

The genetic polymorphism value of each primer was determined and ranged between (46-85%), primer R03 produced the highest percent of genetic polymorphism compared with primer A013.

It was also possible to find the DNA fingerprinting of sixteen of T.

rubrum isolates through the appearance of number of bands that were unique to each isolate.

That may be used in the future to design the detection primer for these isolates directly.

التخصصات الرئيسية

الأحياء

الموضوعات

• الفطريات
• العراق
• الجلد

عدد الصفحات

102

قائمة المحتويات

• APA
• MLA
• AMA

لغة النص

الإنجليزية

نوع البيانات

رسائل جامعية

رقم السجل

BIM-601240