The hereditary cataract is one of the most common eye disease in dogs. The expansion of this disease in the Staffordshire bullterrier breed has been so massive that in the Czech Republic was introduced the rule of mandatory testing of at least one of a breeding pair. This is a degenerative disease of the lens causing total blindness of the affected animal within three years. Since some time ago there are no more dogs affected by the disease in the Czech Republic, there are however still hidden carriers which need to be discovered to the complete extinction of the disease in the genome. The goal of this study was to test simple ways of collecting biological samples, try them in practice and to verify whether they are suitable for the DNA isolation and also to test an alternative method of molecular detection of this disease. In total there have been 23 buccal swabs collected from male and female Staffordshire Bullterrier examples. The detection of the hidden carriers of the hereditary cataract was carried out by PCR analysis with specific primers. The obtained amplicons were detected by both gel and chip electrophoresis and by using fragment analysis. This detection of the carriers was based on the presence of two amplicons (heterozygotes). I came to conclusion that to detect hidden carriers it is neccessary to use the fragment analysis because of the difference of only one base in the reference section of DNA. Neither gel nor chip electrophoresis does provide sufficiently high resolution and it is not possible to detect two fragments that differ only by one bp. As the most appropriate sampling method I have chosen the buccal smear by cytological brush followed by isolating the DNA by Chelex with purification of the sample subsequently.
| Identifer | oai:union.ndltd.org:nusl.cz/oai:invenio.nusl.cz:203393 |
| Date | January 2015 |
| Creators | FARKOVÁ, Barbora |
| Source Sets | Czech ETDs |
| Language | Czech |
| Detected Language | English |
| Type | info:eu-repo/semantics/masterThesis |
| Rights | info:eu-repo/semantics/restrictedAccess |
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