Fanconi anemia diagnostic study of choice

Jump to navigation Jump to search

Fanconi anemia Microchapters


Patient Information


Historical Perspective




Differentiating Fanconi anemia from other Diseases

Epidemiology and Demographics

Risk Factors


Natural History, Complications and Prognosis


Diagnostic Study of Choice

History and Symptoms

Physical Examination

Laboratory Findings


X Ray




Other Imaging Findings

Other Diagnostic Studies


Medical Therapy


Primary Prevention

Secondary Prevention

Cost-Effectiveness of Therapy

Future or Investigational Therapies

Case Studies

Case #1

Fanconi anemia diagnostic study of choice On the Web

Most recent articles

Most cited articles

Review articles

CME Programs

Powerpoint slides


American Roentgen Ray Society Images of Fanconi anemia diagnostic study of choice

All Images
Echo & Ultrasound
CT Images

Ongoing Trials at Clinical

US National Guidelines Clearinghouse

NICE Guidance

FDA on Fanconi anemia diagnostic study of choice

CDC on Fanconi anemia diagnostic study of choice

Fanconi anemia diagnostic study of choice in the news

Blogs on Fanconi anemia diagnostic study of choice

Directions to Hospitals Treating Fanconi anemia

Risk calculators and risk factors for Fanconi anemia diagnostic study of choice

Editor-In-Chief: C. Michael Gibson, M.S., M.D. [1] Associate Editor(s)-in-Chief: Shyam Patel [2]


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] Mutation analysis in Fanconi anemia can also be done by multiplex ligation-dependent probe amplification and PCR-based Sanger sequencing.[4]


  1. 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.
  2. 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. 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.
  4. Gille JJ, Floor K, Kerkhoven L, Ameziane N, Joenje H, de Winter JP (2012). "Diagnosis of Fanconi Anemia: Mutation Analysis by Multiplex Ligation-Dependent Probe Amplification and PCR-Based Sanger Sequencing". Anemia. 2012: 603253. doi:10.1155/2012/603253. PMC 3388349. PMID 22778927.