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

Dissertant

Salman, Zaynab Anas

Thesis advisor

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

University

University of Baghdad

Faculty

College of Science

Department

Biotechnology Department

University Country

Iraq

Degree

Master

Degree Date

2013

English Abstract

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.

Main Subjects

Biology

Topics

No. of Pages

102

Table of Contents

Table of contents.

Abstract.

Abstract in Arabic.

Introdaction.

Chapter One : Literature review.

Chapter Two : Materials and methods.

Chapter Three : Results and discussions.

Conclusions and recommendations.

References.

American Psychological Association (APA)

Salman, Zaynab Anas. (2013). Identification of trichophyton rubrum isolates by using traditional methods and RAPD-PCR. (Master's theses Theses and Dissertations Master). University of Baghdad, Iraq
https://search.emarefa.net/detail/BIM-601240

Modern Language Association (MLA)

Salman, Zaynab Anas. Identification of trichophyton rubrum isolates by using traditional methods and RAPD-PCR. (Master's theses Theses and Dissertations Master). University of Baghdad. (2013).
https://search.emarefa.net/detail/BIM-601240

American Medical Association (AMA)

Salman, Zaynab Anas. (2013). Identification of trichophyton rubrum isolates by using traditional methods and RAPD-PCR. (Master's theses Theses and Dissertations Master). University of Baghdad, Iraq
https://search.emarefa.net/detail/BIM-601240

Language

English

Data Type

Arab Theses

Record ID

BIM-601240