Cancer FISH B-Acute Lymphoblastic Leukemia (ALL) FISH

B-Acute Lymphoblastic Leukemia (ALL) is a disorder of B lymphoblasts that accounts for the vast majority of children with acute leukemia and a fraction of adults with acute leukemia.  B-ALL is broadly categorized as either having recurrent genetic abnormalities or not; recurrent genetic abnormalities include both balanced translocations and aneuploidy (Swerdlow et al. 2008).  This FISH panel is designed to detect the most common, and/or prognostically-significant abnormalities in B-ALL (Swerdlow et al. 2008).  FISH studies are useful adjuncts to complete chromosome studies, particularly when following an abnormal clone, assessing relapse and progression, or when material is inadequate for chromosomal analysis.

The B-ALL FISH panel includes:

1. ETV6/RUNX1 (aka TEL/AML1) dual-fusion probe, to detect t(12;21) and variants; this translocation is cryptic by chromosome analysis.
2. BCR/ABL + ASS dual-fusion probe, to detect t(9;22) and variants.
3. MLL break-apart probe, to detect MLL (11q23.1-11q23.3) rearrangement.
4. Wolf-Hirschhorn (4p16.3)/(4q12)/CEP10(SA) as enumeration probes, to detect hyperdiploidy involving trisomies of chromosomes 4 and 10.
5. E2A/PBX1 dual-fusion probe, to detect t(1;19) and variants.
6. IGH break-apart probe, to detect IGH (14q32) rearrangement.
7. CDKN2A (p16) / CEP 9 to detect loss of 9p21 region.
8. CRLF2 break-apart probe, to detect CRLF2 (Xp22.33/Yp11.32) rearrangement.

Note: Reflex testing may include IL3 break-apart, MLLT3/MLL t(9;11), MLL/MLLT1 t(11;19), ETV6 break-apart, and/or AFF1 break-apart when relevant. Additional charges apply. Interphase FISH analysis is not intended to stand alone, but rather to provide supplemental information to routine cytogenetic studies.

Additional and reflex testing is also available upon provider request for Ph-like B-ALL conditions.  These probes would include ABL1 break apart, ABL2 break apart, JAK2 break apart, and PDGFRB break apart. 

Slides are prepared per standard protocols and 100-200 interphase cells are scored per probe.

 *Please note: it is our laboratory policy to analyze at least one metaphase cell from every hybridization, if possible, including both normal and abnormal cells, if both exist.

• FFPE Slides: Specimen Requirements:
  • If sending slides, please include H&E stained slide cut from same block
  • Preferred slice thickness is 4-5 microns on positively charged slides. 
  • Please submit 12-18 slides.  Store at room temperature.
  • Contact Client Services at (855) 535-1522 for shipping supplies and instructions
• Bone Marrow:  Shortly before aspiration add 0.2 cc of Sodium Heparin (1,000 units/ml) to tube of transport medium (please contact lab to arrange for transport medium to be sent).  Add at least 1 cc of bone marrow aspirate to the tube and suspend well.  Allow no clots to form.
• Bone marrow smears:  Provide 10 slides
• Blood: May be used if bone marrow is inaspirable and blasts (.5%) are present.  Send in Sodium Heparin tube.
  • Adult - 3-5 mL drawn into a GREEN top sodium heparin vacutainer tube or into a pre-heparinized plastic syringe (use 0.2 cc sodium heparin, 1000 unit/mL).  Do NOT use lithium heparin.
  • Child - 1-2 mL, as above.
  • Infant - 1-2 mL, as above. 
  • Keep at room temperature and transport to laboratory as soon as possible.
  • Contact Client Services at (855) 535-1522 for supplies and instructions.
• Bone core biopsy: May also provide cells in cases where marrow is severely packed.  Send in transport medium with Sodium Heparin.
• Lymph node or tumor: Tumor specimens of at least 0.5 cm³ (up to 3 inches in diameter) are immediately collected with sterile methods into closable containers with sterile transport medium.* Needle biopsies will be accepted, but are often difficult to grow.  Deliver the specimen in transport medium to the laboratory within a day, if possible, with decreased success rates as specimens are delayed in transit.  Protect sample from temperature extremes during shipping.

*Sterile Ringer’s solution, either lactated or non-lactated and sterile isotonic saline are alternatives, if no complete RPMI is available.  Contact Client Services for more information on media requirements.

It is preferable to collect the specimen before initiation of chemotherapy in the patient.  Tumor samples should be selected from viable areas, with as little normal or necrotic material as possible. 

Unacceptable specimens are acellular, necrotic specimens, septic specimens, specimens in fixative or frozen, or specimens collected more than one week previously.

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

Mon-Sat

5 - 10 days Contact lab at 855-KDL-1LAB (535-1522) - Time varies depending on number of tests performed.

• Keep specimen cold during transit, but do not ship on dry ice.  
• Please use the cold pack provided in the KDL shipping kit.  
• Ship the specimen overnight express, using the FedEx priority overnight label provided.  
• The specimen must arrive at the lab no more than 24 hours after collection.

1. Swerdlow et al. (Eds.): WHO Classification of Tumors of Haematopoietic and Lymphoid Tissues. IARC: Lyon 2008

2. De Braekeleer et al. 2012 Leuk Res 36, doi: 10.1016/j.leukres.2012.04.010; Haferlach et al. 2012 Genes Chrom Cancer 51, doi: 10.1002/gcc.21918