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==Overview== | ==Overview== | ||
Fanconi anemia | There are two major diagnostic studies of choice for Fanconi anemia. These include chromosomal breakage analysis and mutational analysis. | ||
==Diagnostic study of choice== | ==Diagnostic study of choice== | ||
There are two diagnostic studies of choice for Fanconi anemia: | There are two diagnostic studies of choice for Fanconi anemia: | ||
*'''Chromosomal breakage analysis''': Analysis of DNA susceptibility to DNA-damaging agents has been the traditional diagnostic study of choice for Fanconi anemia for many years. This assay is based on the idea that agents such as diepoxybutane and mitomycin C can create disruptions in chromosomes of cells that have impaired DNA damage response.<ref name="pmid17105750">{{cite journal| author=Antonio Casado J, Callén E, Jacome A, Río P, Castella M, Lobitz S et al.| title=A comprehensive strategy for the subtyping of patients with Fanconi anaemia: conclusions from the Spanish Fanconi Anemia Research Network. | journal=J Med Genet | year= 2007 | volume= 44 | issue= 4 | pages= 241-9 | pmid=17105750 | doi=10.1136/jmg.2006.044719 | pmc=2598052 | url=https://www.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&tool=sumsearch.org/cite&retmode=ref&cmd=prlinks&id=17105750 }} </ref> In patients with Fanconi anemia, their cells demonstrate hypersensitivity to these DNA-damaging agents because the cells already harbor mutation in genes (like ''FANC'' family genes) which normally allow for correction of DNA damage. | *'''Chromosomal breakage analysis''': Analysis of DNA susceptibility to DNA-damaging agents has been the traditional diagnostic study of choice for Fanconi anemia for many years. This assay is based on the idea that agents such as diepoxybutane and mitomycin C can create disruptions in chromosomes of cells that have impaired DNA damage response.<ref name="pmid17105750">{{cite journal| author=Antonio Casado J, Callén E, Jacome A, Río P, Castella M, Lobitz S et al.| title=A comprehensive strategy for the subtyping of patients with Fanconi anaemia: conclusions from the Spanish Fanconi Anemia Research Network. | journal=J Med Genet | year= 2007 | volume= 44 | issue= 4 | pages= 241-9 | pmid=17105750 | doi=10.1136/jmg.2006.044719 | pmc=2598052 | url=https://www.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&tool=sumsearch.org/cite&retmode=ref&cmd=prlinks&id=17105750 }} </ref> These agents crosslink DNA.<ref name="pmid19622403">{{cite journal| author=Auerbach AD| title=Fanconi anemia and its diagnosis. | journal=Mutat Res | year= 2009 | volume= 668 | issue= 1-2 | pages= 4-10 | pmid=19622403 | doi=10.1016/j.mrfmmm.2009.01.013 | pmc=2742943 | url=https://www.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&tool=sumsearch.org/cite&retmode=ref&cmd=prlinks&id=19622403 }} </ref> In patients with Fanconi anemia, their cells demonstrate hypersensitivity to these DNA-damaging agents because the cells already harbor mutation in genes (like ''FANC'' family genes) which normally allow for correction of DNA damage. | ||
**''Methods'': In the chromosomal breakage test, peripheral blood lymphocytes are isolated then stimulated with phytohemagglutinin (PHA) for 24 hours. Cells are then treated with either diepoxybutane or mitomycin C for 48 hours. Aberrant metaphases are visualized via microscopy, with specific attention to chromosome breaks or radial chromosomes.<ref name="pmid17105750">{{cite journal| author=Antonio Casado J, Callén E, Jacome A, Río P, Castella M, Lobitz S et al.| title=A comprehensive strategy for the subtyping of patients with Fanconi anaemia: conclusions from the Spanish Fanconi Anemia Research Network. | journal=J Med Genet | year= 2007 | volume= 44 | issue= 4 | pages= 241-9 | pmid=17105750 | doi=10.1136/jmg.2006.044719 | pmc=2598052 | url=https://www.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&tool=sumsearch.org/cite&retmode=ref&cmd=prlinks&id=17105750 }} </ref> | **''Methods'': In the chromosomal breakage test, peripheral blood lymphocytes are isolated then stimulated with phytohemagglutinin (PHA) for 24 hours. Cells are then treated with either diepoxybutane or mitomycin C for 48 hours. Aberrant metaphases are visualized via microscopy, with specific attention to chromosome breaks or radial chromosomes.<ref name="pmid17105750">{{cite journal| author=Antonio Casado J, Callén E, Jacome A, Río P, Castella M, Lobitz S et al.| title=A comprehensive strategy for the subtyping of patients with Fanconi anaemia: conclusions from the Spanish Fanconi Anemia Research Network. | journal=J Med Genet | year= 2007 | volume= 44 | issue= 4 | pages= 241-9 | pmid=17105750 | doi=10.1136/jmg.2006.044719 | pmc=2598052 | url=https://www.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&tool=sumsearch.org/cite&retmode=ref&cmd=prlinks&id=17105750 }} </ref> | ||
*'''Mutational analysis''': In the recent years, DNA sequencing has become widely available, and diagnosis of Fanconi anemia can be made by sequencing of candidate genes known to be involved in the pathogenesis of Fanconi anemia. These genes mostly include members of the ''FANC'' family.<ref name="pmid29580219">{{cite journal| author=Hebert PDN, Braukmann TWA, Prosser SWJ, Ratnasingham S, deWaard JR, Ivanova NV et al.| title=A Sequel to Sanger: amplicon sequencing that scales. | journal=BMC Genomics | year= 2018 | volume= 19 | issue= 1 | pages= 219 | pmid=29580219 | doi=10.1186/s12864-018-4611-3 | pmc=5870082 | url=https://www.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&tool=sumsearch.org/cite&retmode=ref&cmd=prlinks&id=29580219 }} </ref> | *'''Mutational analysis''': In the recent years, DNA sequencing has become widely available, and diagnosis of Fanconi anemia can be made by sequencing of candidate genes known to be involved in the pathogenesis of Fanconi anemia. These genes mostly include members of the ''FANC'' family.<ref name="pmid29580219">{{cite journal| author=Hebert PDN, Braukmann TWA, Prosser SWJ, Ratnasingham S, deWaard JR, Ivanova NV et al.| title=A Sequel to Sanger: amplicon sequencing that scales. | journal=BMC Genomics | year= 2018 | volume= 19 | issue= 1 | pages= 219 | pmid=29580219 | doi=10.1186/s12864-018-4611-3 | pmc=5870082 | url=https://www.ncbi.nlm.nih.gov/entrez/eutils/elink.fcgi?dbfrom=pubmed&tool=sumsearch.org/cite&retmode=ref&cmd=prlinks&id=29580219 }} </ref> | ||
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Editor-In-Chief: C. Michael Gibson, M.S., M.D. [1] Associate Editor(s)-in-Chief: Shyam Patel [2]
Overview
There are two major diagnostic studies of choice for Fanconi anemia. These include chromosomal breakage analysis and mutational analysis.
Diagnostic study of choice
There are two diagnostic studies of choice for Fanconi anemia:
- Chromosomal breakage analysis: Analysis of DNA susceptibility to DNA-damaging agents has been the traditional diagnostic study of choice for Fanconi anemia for many years. This assay is based on the idea that agents such as diepoxybutane and mitomycin C can create disruptions in chromosomes of cells that have impaired DNA damage response.[1] These agents crosslink DNA.[2] In patients with Fanconi anemia, their cells demonstrate hypersensitivity to these DNA-damaging agents because the cells already harbor mutation in genes (like FANC family genes) which normally allow for correction of DNA damage.
- Methods: In the chromosomal breakage test, peripheral blood lymphocytes are isolated then stimulated with phytohemagglutinin (PHA) for 24 hours. Cells are then treated with either diepoxybutane or mitomycin C for 48 hours. Aberrant metaphases are visualized via microscopy, with specific attention to chromosome breaks or radial chromosomes.[1]
- Mutational analysis: In the recent years, DNA sequencing has become widely available, and diagnosis of Fanconi anemia can be made by sequencing of candidate genes known to be involved in the pathogenesis of Fanconi anemia. These genes mostly include members of the FANC family.[3]
- Methods: Mutational analysis can be done through a variety of methods. In the most simple assay, mutational analysis begins with isolation of DNA from peripheral blood lymphocytes. Mutation-specific primers for FANC family genes are designed, and a polymerase chain reaction (PCR) is performed. The visualization of bands corresponding to an amplicon containing the mutation would confirm that that a mutation exists. In more recently developed assays, automated sequencing based on Sanger methods allow for precise determination of mutation status.[3]
References
- ↑ 1.0 1.1 Antonio Casado J, Callén E, Jacome A, Río P, Castella M, Lobitz S; et al. (2007). "A comprehensive strategy for the subtyping of patients with Fanconi anaemia: conclusions from the Spanish Fanconi Anemia Research Network". J Med Genet. 44 (4): 241–9. doi:10.1136/jmg.2006.044719. PMC 2598052. PMID 17105750.
- ↑ Auerbach AD (2009). "Fanconi anemia and its diagnosis". Mutat Res. 668 (1–2): 4–10. doi:10.1016/j.mrfmmm.2009.01.013. PMC 2742943. PMID 19622403.
- ↑ 3.0 3.1 Hebert PDN, Braukmann TWA, Prosser SWJ, Ratnasingham S, deWaard JR, Ivanova NV; et al. (2018). "A Sequel to Sanger: amplicon sequencing that scales". BMC Genomics. 19 (1): 219. doi:10.1186/s12864-018-4611-3. PMC 5870082. PMID 29580219.