| Single cell analysis of mutations in the APC gene. | |
| | |
MedLine Citation:
|
PMID: 19828935 Owner: NLM Status: In-Process |
Abstract/OtherAbstract:
|
INTRODUCTION: Mutation analysis of inherited monogenic diseases is an important aspect of preimplantation genetic diagnosis. Familial adenomatous polyposis of the colon is an autosomal dominant inherited disorder caused by mutations in the tumor suppressor gene adenomatous polyposis coli (APC). A characteristic of this severe disease is the development of thousands of polyps in the colon which untreated evolve into malignant colon carcinomas. Here we present a novel approach for the establishment of mutation detection in the APC gene in single cells applicable for preimplantation genetic diagnosis. METHODS: Single fixed lymphocytes were isolated via laser microdissection and transferred to reaction centers of planar chemically structured glass slides via a recently developed horizontal low-pressure single particle adsorbing transfer system (SPATS). A multiplex nested polymerase chain reaction protocol in a 1-microl reaction volume, followed by sequencing and fragment length analysis was applied in order to detect mutations in the APC gene. RESULTS: Reliable isolation and transfer of single lymphocytes was demonstrated. High amplification efficiency and low allelic drop out (ADO) rates for polymorphic microsatellite markers and mutation specific amplification products of various mutations in the APC gene were obtained from fixed single cells. CONCLUSIONS: This novel nanotechnological downscaling approach enables a reliable validation of genetic testing using diploid single lymphocytes, and will open a wide range of single cell diagnostics. |
| | |
Authors:
|
Veronika Mayer; Ulrike Schoen; Elke Holinski-Feder; Udo Koehler; Stefan Thalhammer |
Related Documents
:
|
16478485 - Molecular characterization of the bacteria adherent to human colorectal mucosa. 15365995 - Germline mutations in mlh1, msh2 and msh6 in korean hereditary non-polyposis colorectal... 15613555 - Genetic characterization of chinese hereditary non-polyposis colorectal cancer by dhplc... 10911905 - Molecular genetics of hereditary nonpolyposis colorectal cancer. 16719905 - Association study of genetic variants of pro-inflammatory chemokine and cytokine genes ... 12119565 - Premature ovarian failure and fmr1 premutation co-segregation in a large brazilian family. |
Publication Detail:
|
Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2009-10-13 |
Journal Detail:
|
Title: Fetal diagnosis and therapy Volume: 26 ISSN: 1421-9964 ISO Abbreviation: Fetal. Diagn. Ther. Publication Date: 2009 |
Date Detail:
|
Created Date: 2009-12-09 Completed Date: - Revised Date: - |
Medline Journal Info:
|
Nlm Unique ID: 9107463 Medline TA: Fetal Diagn Ther Country: Switzerland |
Other Details:
|
Languages: eng Pagination: 148-56 Citation Subset: IM |
Affiliation:
|
Helmholtz Zentrum Munich, German Research Center for Environmental Health, Institute of Radiation Protection, Neuherberg, Germany. |
Export Citation:
|
APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
|
|
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
Previous Document: Development of the performance measure for activities of daily living-8 for patients with congestive...
Next Document: The Elderly Patient and Cardiac Surgery - A Mini-Review.