Terminal regions of chromosomes are frequently involved in structural chromosomal abnormalities and are prone to rearrangements. They are also gene rich, and have been shown to be associated with a number of clinical conditions. This study focuses on telomere-specific fluorescence in situ hybridization (FISH) probes. It addresses the technical aspects of their preparation and application, and examines whether FISH using these probes is a valuable tool for detection of aberrations in the terminal regions of chromosomes. A set of telomere specific probes was generated from half yeast artificial chromosome (YAC) and cosmid clones by DNA preparation via Alu-PCR, alkaline lysis, cesium chloride-ethidium bromide centrifugation, or using a "Qiagen" kit. Probe quality and optimal FISH conditions were established by hybridizing half-YACs specific to telomeres 21q, 18q, 10p, and cosmids for telomeres 2q/8p, 13q, 14q and 20p to normal metaphases. Probes for 10p, 13q, 14q, 18q and 21q yielded clear and specific hybridization signals, present in over 90% of metaphases analyzed. Interphase analysis using the probes was not accurate. The ability of the telomeric probes to characterize balanced and unbalanced abnormalities was established by hybridization to patients with previously diagnosed chromosomal aberrations. The probes identified and confirmed partial trisomies 21q, 18q, and balanced translocations t(8;14), t(6;14), t(8;18), t(5;10). In three of the cases, partial monosomy 18q, translocation t(10;13), and t(1;10), the probes were able to provide information which was not revealed by banding analysis. This study demonstrates the ability of telomere specific probes to characterize aberrations in the terminal regions of chromosomes, and concludes that FISH using the telomeric probes is a valuable tool for clinical cytogenetics.
| Identifer | oai:union.ndltd.org:LACETR/oai:collectionscanada.gc.ca:QMM.27497 |
| Date | January 1997 |
| Creators | Bielanska, Magdalena M. |
| Contributors | Eydoux, P. (advisor) |
| Publisher | McGill University |
| Source Sets | Library and Archives Canada ETDs Repository / Centre d'archives des thèses électroniques de Bibliothèque et Archives Canada |
| Language | English |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Format | application/pdf |
| Coverage | Master of Science (Department of Pathology.) |
| Rights | All items in eScholarship@McGill are protected by copyright with all rights reserved unless otherwise indicated. |
| Relation | alephsysno: 001601423, proquestno: MQ37095, Theses scanned by UMI/ProQuest. |
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