• Test Code:
  • Department:
  • Test Synonyms:
    Histone H3.3
  • CPT Code(s):

H3F3A encodes histone H3.3. Mutations in this gene occur in 10-15% of centrally located high grade gliomas and glioblastomas, and are even more common in pediatric tumors, particularly in diffuse intrinsic pontine gliomas. Mutations of codon K28 (also referred to as K27 in the literature) are associated with a shorter survival time compared to non-mutated tumors, whereas those with mutations of codon G34 show better survival.1-2


After review by a pathologist, tumor-rich areas are macro-dissected from FFPE sections and DNA is extracted and purified. Exon 1 of the H3F3A gene (including codons K28 and G34) is amplified by PCR and the product is subjected to bi-directional Sanger sequencing. The estimated sensitivity of this method is approximately 20% mutant allele.

Specimen Requirements:

  • Unstained FFPE slides at room temperature.
  • Preferred slice thickness is 5 micrometer on positively charged slides.
  • Please submit 10 unstained slides; 15 sections for small biopsies.
  • Contact Client Services with questions.
  • Tumor samples should be selected from viable areas, with as little normal or necrotic material as possible.
  • After collection, keep at room temperature until shipping.
  • Contact Client Services at (855) 535-1522 for shipping kits and instructions.

A REQUISITION FORM MUST ACCOMPANY ALL SAMPLES. Please include detailed clinical information.​

Test Performed (Days):


Turn Around Time:

5 Days - Contact Lab at 855-KDL-1LAb (535-1522)

Shipment Sensitivity Requirements:

  • Keep specimen at room temperature during transit.
  • Do not use the cold pack provided in the KDL shipping kit.
  • Ship the specimen overnight express, using the FedEx priority overnight label provided. 


  1. Sturm D, Witt H, Hovestadt V et al Hotspot mutations in H3F3A and IDH1 define distinct epigenetic and biological subgroups of glioblastoma. Cancer Cell. 2012 Oct 16, 22(4):425-437.
  2. Gielen GH, Gessi M, Hammes J, Kramm CM, Waha A, Pietsch T. H3F3A K27M mutation in pediatric CNS umors: a marker for diffuse high-grade astrocytomas. Am J Clin Pathol. 2013 Mar, 139 (3):345-349.

Additional Info:

The Knight Cancer Institute at Oregon Health & Science University is a pioneer in the field of precision cancer medicine. The institute's director, Brian Druker, M.D., helped prove it was possible to shut down just the cells that enable cancer to grow. This breakthrough has made once-fatal forms of the disease manageable and transformed how cancer is treated. The OHSU Knight Cancer Institute is the only National Cancer Institute-designated Cancer Center between Sacramento and Seattle – an honor earned only by the nation's top cancer centers. It is headquarters for one of the National Cancer Institute's largest research collaboratives, SWOG, in addition to offering the latest treatments and technologies as well as hundreds of research studies and clinical trials.

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