Somatic Hypermutation Analysis, IGH, Sequencing

Indication

Detection of the rate of mutation in the expressed IGH variable region of the B-cell tumor clone for use in prognostication between pre-germinal center and post-germinal center subsets of chronic lymphocytic leukemia.

Methodology

RNA is extracted from white blood cells in bone marrow, and/or peripheral blood and reverse transcribed to cDNA. Multiplex PCR amplification of IGH transcripts is done using consensus VH primers. DNA sequencing of the PCR product is done and compared to germline IGH sequences in the EMBL database.

Sensitivity

Requires that the B-cell clone comprise at least 10% of the leukocytes in the sample. Some samples with adequate clonal B-cells (approximately 10%) will be uninterpretable due to mutations in the primer binding sequences.

Turnaround Time

Five to 10 working days

Sample Requirements

• 10-30 ml peripheral blood in lavender top (EDTA) tube, sent on wet ice
  
  or
  
• 2-5 ml of bone marrow aspirate in lavender top (EDTA) tube, sent on wet ice
  
  or
  
• 15 µg of purified RNA, sent on dry ice
  
  or
  
• 10 µg of cDNA, sent on dry ice

CPT Codes

81263

The CPT codes provided are based on AMA guidelines and are for informational purposes only. CPT coding is the sole responsibility of the billing party. Please direct any questions regarding coding to the payer being billed.