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Evaluation Of Risk Factors And Molecular Diagnosis Of Dermatophytosis In Dogs

By: Muhammad Haseeb Saeed (2008-VA-241) | Prof. Dr. Aneela Zameer Durrani.
Contributor(s): Dr. Hassan Saleem | Prof. Dr. Azhar Maqbool.
Material type: materialTypeLabelBookPublisher: 2016Description: 29p.Subject(s): Department of Clinical Medicine | CMSDDC classification: 2528-T Dissertation note: Dogs are most kept and beloved pets in Pakistani society. Dermatophytosis is among the common disease of the pets. Many predisposing factors are involved in development of clinical cases of dermatophytosis including climatic conditions, housing condition of dogs and physical attributes such as coat hair size. Dermatophytosis is not only of concern as being infection of pets but also of its zoonotic importance hence it is very crucial to diagnose dermatophytic infection well in time. Dermatophytosis is caused by Dermatophytes,Microsporum, Trichophyton and Epidermophyton, the fungal species. It is difficult to diagnose the Dermatophytosis from other skin infections by routine tests in most of the cases especially subclinical. Polymerase Chain Reaction (PCR) is advanced and the most reliable technique to detect genome of Dermatophytes even in minute quantities specifically and can efficiently detect the presence of any Dermatophyte specie on the skin of dog. The current study was planned to develop and validate a diagnostic assay which could be able to detect and distinguish tree important dermatophytes species including Microsporum, Trichophyton and Epidermophytonby a uniplex PCR reaction. Analysis of involvement of certain predisposing factors in dermatophytosis was second goal to be worked on in this study. Samples of suspected pet dogs (n=50) were collected by scraping the skin at affected areas over skin. DNA was extracted from the skin scraping samples by organic Phenol Chloroform Isoamyle Alcohol method. Primers, specific to the 18-S ribosomal RNA region of genomes of the Dermatophytes, were designed after alignment of available sequences of Microsporum,Trichophyton and Epidermophyton at NCBI. Annealing temperature and recipe of PCR reaction was optimized by gradient PCR in BIO-Rad thermal cycler. Amplification reaction of all samples collected was carried out as per optimized reaction conditions, afterwards. Amplified products obtained were subjected to genotyping by agarose gel electrophoresis for size based separation of the amplified products. The specific amplified bands of desired genomic region of dermatophytes were seen in UV light transilluminator. The data of results of predisposing factors involved in dermatophytosis wasanalysedby using Pearson’s chi squared test with the help of Statistical Package for the Social Science (SPSS) Program. Genome specific product sizes of Microsporum and Trichophyton i.e. 366 bp and 351 bp in respective positive samples were observed. Out of 50 suspected samples 46 samples were positive for dermatophytosis out of which 38 samples (82.6%) were positive for Microsporum, 6 samples (13%) for Trichophyton and 2 samples (4.4%) were positive for both Microsporumand Trichophyton. This study will help to validate a diagnostic technique for Dermatophytosis with greater efficacy and reliability. Moreover, this investigation may become basis for the future research activities in this field in Pakistan.
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Item type Current location Collection Call number Status Date due Barcode Item holds
Thesis Thesis UVAS Library
Thesis Section
Veterinary Science 2528-T (Browse shelf) Available 2528-T
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Dogs are most kept and beloved pets in Pakistani society. Dermatophytosis is among the common disease of the pets. Many predisposing factors are involved in development of clinical cases of dermatophytosis including climatic conditions, housing condition of dogs and physical attributes such as coat hair size. Dermatophytosis is not only of concern as being infection of pets but also of its zoonotic importance hence it is very crucial to diagnose dermatophytic infection well in time. Dermatophytosis is caused by Dermatophytes,Microsporum, Trichophyton and Epidermophyton, the fungal species. It is difficult to diagnose the Dermatophytosis from other skin infections by routine tests in most of the cases especially subclinical. Polymerase Chain Reaction (PCR) is advanced and the most reliable technique to detect genome of Dermatophytes even in minute quantities specifically and can efficiently detect the presence of any Dermatophyte specie on the skin of dog. The current study was planned to develop and validate a diagnostic assay which could be able to detect and distinguish tree important dermatophytes species including Microsporum, Trichophyton and Epidermophytonby a uniplex PCR reaction. Analysis of involvement of certain predisposing factors in dermatophytosis was second goal to be worked on in this study. Samples of suspected pet dogs (n=50) were collected by scraping the skin at affected areas over skin. DNA was extracted from the skin scraping samples by organic Phenol Chloroform Isoamyle Alcohol method. Primers, specific to the 18-S ribosomal RNA region of genomes of the Dermatophytes, were designed after alignment of available sequences of Microsporum,Trichophyton and Epidermophyton at NCBI. Annealing temperature and recipe of PCR reaction was optimized by gradient PCR in BIO-Rad thermal cycler. Amplification reaction of all samples collected was carried out as per optimized reaction conditions, afterwards. Amplified products obtained were subjected to genotyping by agarose gel electrophoresis for size based separation of the amplified products. The specific amplified bands of desired genomic region of dermatophytes were seen in UV light transilluminator. The data of results of predisposing factors involved in dermatophytosis wasanalysedby using Pearson’s chi squared test with the help of Statistical Package for the Social Science (SPSS) Program.
Genome specific product sizes of Microsporum and Trichophyton i.e. 366 bp and 351 bp in respective positive samples were observed. Out of 50 suspected samples 46 samples were positive for dermatophytosis out of which 38 samples (82.6%) were positive for Microsporum, 6 samples (13%) for Trichophyton and 2 samples (4.4%) were positive for both Microsporumand Trichophyton.
This study will help to validate a diagnostic technique for Dermatophytosis with greater efficacy and reliability. Moreover, this investigation may become basis for the future research activities in this field in Pakistan.

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