FISH and PCR Testing for AML, ALL, CML, MM, CLL, and MDS

Andy Nguyen, M.D. / UT-Medical School at Houston, Pathology/ Last Revision on: 7/25/2012

1. AML

(a) Sample in green-top tube for

-Cytogenetics

-AML Profile, FISH

-BCR/ABL by FISH

***Note that if FISH result is positive for BCR/ABL or PML/RARA, send for quantitative PCR for the same mutation in follow-up bone marrows (sample in lavender-top tube)

(b) Sample in lavender-top tube for PCR testing for

-FLT3 (ITD) and D835 Variant

-NPM1

-CEBPA

2. ALL

Sample in green-top tube for

-Cytogenetics

-ALL Profile, FISH

3. CML

(a) Sample in green-top tube for

-Cytogenetics

-BCR/ABL by FISH

(b) For follow-up bone marrow, PCR-quantitative for bcr-abl1 (sample in lavender-top tube)

(c) For detection of resistance to Imatinib Mesylate: Bcr-abl1 Kinase Domain Mutation Analysis (sample in purple-top tube for PCR)

4. Myeloid and Lymphoid Neoplasms with Eosinophilia

Sample in green-top tube for

-Cytogenetics

-FISH testing for:

-FIP1L1/PDGFRA

-PDGFRB

-FGFR1

5. PLASMA CELL MYELOMA

Green top tube for

-Cytogenetics

-Multiple Myeloma Profile, FISH

6. CLL

Green top tube for

-Cytogenetics

-CLL Profile, FISH

7. MDS

Green top tube for

-Cytogenetics

-MDS Profile, FISH

8. AVAILABLE FISH TESTING FOR LYMPHOMA (unstained slides or paraffin block)

t(11;14): IgH/bcl1, mantle cell lymphoma

MYC: 8q24, Burkitt lymphoma

t(14;18): IgH/blc2, follicular lymphoma

bcl6: 3q27, DLBCL

MALT1: 18q21, MALT lymphoma

ALK: 2p23, ALTCL-ALK positive

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MORE DETAILS FOR LISTING ABOVE

1. FISH TESTING FOR ACUTE MYELOID LEUKEMIA (sample in green-top tube)

Besides cytogenetics, the following FISH tests are to be sent:

t(15;17) : PML-RARA, good prognosis

inv(16) or t(16;16) : CBFB-MYH11, good prognosis

t(8;21) : RUNX1-RUNX1T1, good prognosis

t(9;22) : BCR-ABL1, poor prognosis

11q23: MLL, poor prognosis

Inv(3): 3q26, EVI1, poor prognosis

***Note that if FISH result is positive for BCR/ABL or PML/RARA, send for quantitative PCR for the same mutation in follow-up bone marrows (sample in lavender-top tube)

2. PCR TESTING FOR ACUTE MYELOID LEUKEMIA (sample in lavender-top tube)

Flt3 , poor prognosis

NPM1 , good prognosis

CEBPA , good prognosis

++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++

ALL

FISH TESTING FOR ACUTE LYMPHOBLASTIC LEUKEMIA (sample in green-top tube)

Besides cytogenetics, the following FISH tests are to be sent:

t(9;22) : BCR-ABL1, poor prognosis, Gleevec can be added to chemo

t(12;21): ETV6-RUNX1, good prognosis (not detected with cytogenetics)

11q23: MLL, poor prognosis

++++++++++++++++++++++++++++++++++

CML

-FISH for bcr-abl1 with new diagnosis (diagnostic sample) of CML (sample in green-top tube)

-For follow-up bone marrow, PCR-quantitative for bcr-abl1 (sample in lavender-top tube)

-For detection of resistance to Imatinib Mesylate: Bcr-abl1 Kinase Domain Mutation Analysis (sample in purple-top tube)

+++++++++++++++

Myeloid and Lymphoid Neoplasms with Eosinophilia (sample in green-top tube)

Besides cytogenetics, the following FISH tests are to be sent:

FIP1L1/PDGFRA: 4q12, good response to Gleevec

PDGFRB: 5q32, good response to Gleevec

FGFR1: 8p12, no response to Gleevec

++++++++++++++++++++++++++++++

FISH TESTING FOR PLASMA CELL MYELOMA

Besides cytogenetics, the following FISH tests are to be sent (sample should be in green-top tube):

13q : 13q14.3 deletion, poor prognosis

t(11;14) : IgH/cyclin D1, good prognosis

t(4;14) : IgH/FGFR3, poor prognosis

17p :17p13.1 deletion (p53 gene deletion), poor prognosis

t(14;16) : IgH/CMAF, poor prognosis

++++++++++++++++++++++++++++

FISH TESTING FOR CLL

Besides cytogenetics, the following FISH tests may be sent (sample should be in green-top tube):

Del 13q : probe D13S319, good prognosis

Del 17p: probe p53, poor prognosis

Trisomy 12 : probe CEP (centromere of chromosome 12), poor prognosis

Del 11q: probe ATM, poor prognosis

++++++++++++++++++++

FISH TESTING FOR MDS

Besides cytogenetics, the following FISH tests may be sent (sample should be in green-top tube):

5q: del(5q) with good prognosis

7q: del(7q) with poor prognosis

++++++++++++++++

FISH TESTING FOR LYMPHOMA (unstained slides or paraffin block)

t(11;14): IgH/bcl1, mantle cell lymphoma

MYC: 8q24, Burkitt lymphoma

t(14;18): IgH/blc2, follicular lymphoma

bcl6: 3q27, DLBCL

MALT1: 18q21, MALT lymphoma

ALK: 2p23, ALTCL-ALK positive

+++++++++++++++++++++++++++++++++++++