Test Code HAEV1 Hemolytic Anemia Evaluation, Blood

Ordering Guidance

Preliminary screening tests, such as complete blood cell count with peripheral smear and direct Coombs test with a negative result, should be run before ordering this evaluation.

Cold agglutinin disorders and autoimmune disorders should be excluded prior to testing. This evaluation is not suitable for acquired causes of hemolysis.

Shipping Instructions

Specimens must arrive within 72 hours of collection.

Necessary Information

At minimum, include recent transfusion information and most recent complete blood cell count results.

Metabolic Hematology Patient Information (T810) is strongly recommended. Testing may proceed without this information, however if the information requested is received, any pertinent reported clinical features and data will drive the focus of the evaluation and be considered in the interpretation.

The laboratory has extensive experience in hemoglobin variant identification and many cases can be confidently classified without molecular testing. However, molecular confirmation is always available, subject to sufficient sample quantity (eg, multiplex ligation-dependent probe amplification testing requires at least 2 mL of specimen in addition to protein testing requirements). If no molecular testing or specific molecular tests are desired, utilize the appropriate check boxes on the form. If the form or other communication is not received, the reviewing hematopathologist will select appropriate tests to sufficiently explain the protein findings, which may or may not include molecular testing.

Specimen Required

The following specimens are required for testing:

2 Whole blood EDTA specimens

2 Whole blood ACD specimens

1 EDTA control specimen

2 Well-made peripheral blood smears (Wright stained or fixed in absolute methanol)

Patient:

Specimen Type: Whole blood

Container/Tube: Lavender top (EDTA) and yellow top (ACD)

Specimen Volume:

EDTA: Two 4-mL vials

ACD: Two 6-mL vials

Collection Instructions:

1. Immediately refrigerate specimens after collection.

2. Send whole blood specimens in original tubes. Do not aliquot.

3. Rubber band patient specimen and control vial together.

Specimen Type: Slides

Container/Tube: Blood smears

Specimen Volume: 2 Peripheral blood smears

1. Prepare 2 peripheral blood smears from 1 of the EDTA tubes collected from the patient

2. Either stain the smear with Wright stain or fix the smear with absolute methanol prior to shipping.

Normal Shipping Control:

Specimen Type: Whole blood

Container/Tube: Lavender top (EDTA)

Specimen Volume: 4 mL

Collection Instructions:

1. Collect a control specimen from a normal (healthy), unrelated, nonsmoking person at the same time as the patient.

2. Clearly hand write normal control on the outermost label.

3. Immediately refrigerate specimen after collection.

4. Send specimen in original tube. Do not aliquot.

5. Rubber band patient specimen and control vial together.

Reflex Tests

Test ID	Reporting Name	Available Separately	Always Performed
SDEX	Sickle Solubility, B	Yes	No
IEF	Isoelectric Focusing, B	No	No
MASS	Hb Variant by Mass Spec, B	No	No
HPFH	Hb F Distribution, B	No	No
ATHAL	Alpha-Globin Gene Analysis	Yes	No
WASQR	Alpha Globin Gene Sequencing, B	Yes, (Order WASEQ)	No
WBSQR	Beta Globin Gene Sequencing, B	Yes, (Order WBSEQ)	No
WBDDR	Beta Globin Cluster Locus Del/Dup,B	Yes, (Order WBDD)	No
WGSQR	Gamma Globin Full Gene Sequencing	Yes, (Order WGSEQ)	No
HAEV0	Hemolytic Anemia Summary Interp	No	No

Specimen Type

Control
Whole Blood ACD-B
Whole Blood EDTA
Whole Blood Slide

Specimen Minimum Volume

EDTA Blood: 3 mL
ACD Blood: 5 mL

Specimen Stability Information

Specimen Type	Temperature	Time	Special Container
Control	Refrigerated	72 hours	PURPLE OR PINK TOP/EDTA
Whole Blood ACD-B	Refrigerated	72 hours
Whole Blood EDTA	Refrigerated	72 hours
Whole Blood Slide	Refrigerated		CARTRIDGE

Reject Due To

Gross hemolysis

Reject

Reference Values

Hemoglobin Variant, A2 and F Quantitation

HEMOGLOBIN A

0-30 days: 5.9-77.2%

1-2 months: 7.9-92.4%

3-5 months: 54.7-97.1%

6-8 months: 80.0-98.0%

9-12 months: 86.2-98.0%

13-17 months: 88.8-98.0%

18-23 months: 90.4-98.0%

≥24 months: 95.8-98.0%

HEMOGLOBIN A2

0-30 days: 0.0-2.1%

1-2 months: 0.0-2.6%

3-5 months: 1.3-3.1%

≥6 months: 2.0-3.3%

HEMOGLOBIN F

0-30 days: 22.8-92.0%

1-2 months: 7.6-89.8%

3-5 months: 1.6-42.2%

6-8 months: 0.0-16.7%

9-12 months: 0.0-10.5%

13-17 months: 0.0-7.9%

18-23 months: 0.0-6.3%

≥24 months: 0.0-0.9%

VARIANT 1

0.0

VARIANT 2

0.0

VARIANT 3

0.0

Hemoglobin Stability

Normal (reported as normal [stable] or abnormal [unstable])

OSMOTIC FRAGILITY

≥12 months:

0.50 g/dL NaCl (unincubated): 3-53% hemolysis

0.60 g/dL NaCl (incubated): 14-74% hemolysis

0.65 g/dL NaCl (incubated): 4-40% hemolysis

0.75 g/dL NaCl (incubated): 1-11% hemolysis

NaCl = sodium chloride