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Test Code IABCS B-Cell Phenotyping Profile for Immunodeficiency and Immune Competence Assessment, Blood

Reporting Name

Immune Assessment B Cell Subsets, B

Useful For

Screening for common variable immunodeficiency (CVID) and hyper-IgM syndromes

 

Assessing B-cell subset reconstitution after stem cell or bone marrow transplant

 

Assessing response to B-cell-depleting immunotherapy

 

Identifying defects in transmembrane activator and calcium modulator and cyclophilin ligand (CAML) interactor (TACI) and B-cell-activating factor receptor (BAFF-R) in patients presenting with clinical symptoms and other laboratory features consistent with CVID

Profile Information

Test ID Reporting Name Available Separately Always Performed
TBBS QN Lymphocyte Subsets: T, B, and NK Yes Yes
IABC Immune Assessment B Cell Subsets, B No Yes

Reflex Tests

Test ID Reporting Name Available Separately Always Performed
CVID CVID Confirmation Flow Panel Yes No

Testing Algorithm

If immune assessment B-cell subsets test is abnormal, then confirmation will be performed at an additional charge.

 

When multiple specimen types are required to perform a panel of tests, the laboratory will perform the tests for which the appropriate specimen type was received and the laboratory will cancel those for which the appropriate specimen was not received. Please be advised that this may change the degree of interpretation received with the report. If only the refrigerate EDTA sample is received, this test will be canceled and converted to RBCS / Relative B-Cell Subset Analysis Percentage which provides the relative B-cell subset values without quantitation.

Performing Laboratory

Mayo Clinic Laboratories in Rochester

Specimen Type

Whole Blood EDTA


Advisory Information


 



Shipping Instructions


Specimens are required to be received in the laboratory weekdays and by 4 p.m. on Friday. Draw and package specimen as close to shipping time as possible.

 

It is recommended that specimens arrive within 24 hours of draw.

 

Samples arriving on the weekend and observed holidays may be canceled.



Necessary Information


1. Date of draw is required.

2. Ordering physician's name and phone number are required.



Specimen Required


Two separate EDTA specimens are required: 1 refrigerated and 1 at ambient transport temperature.

 

For serial monitoring, we recommend that specimen draws be performed at the same time of day.

 

Specimen Type: Whole blood for TBBS / Quantitative Lymphocyte Subsets: T, B, and NK

Container/Tube: 4 mL Lavender top (EDTA)

Specimen Volume: 3 mL

Collection Instructions:

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

2. Label specimen as blood for TBBS / Quantitative Lymphocyte Subsets: T, B, and NK.

Specimen Stability Information: Ambient <52 hours

 

Specimen Type: Whole blood for IABC / B-Cell Phenotyping Screen for Immunodeficiency and Immune Competence Assessment, Blood

Container/Tube: Lavender top (EDTA)

Specimen Volume:

≤14 years: 4 mL

>14 years: 10 mL

Collection Instructions:

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

2. Label specimen as blood for IABC / B-Cell Phenotyping Screen for Immunodeficiency and Immune Competence Assessment, Blood.

Specimen Stability Information: Refrigerated <48 hours


Specimen Minimum Volume

TBBS: 1 mL
IABC
≤14 years: 3 mL
>14 years: 5 mL

Specimen Stability Information

Specimen Type Temperature Time Special Container
Whole Blood EDTA Varies 48 hours PURPLE OR PINK TOP/EDTA

Reference Values

The appropriate age-related reference values will be provided on the report.

Day(s) and Time(s) Performed

Monday through Friday

Specimens are required to be received in the laboratory on weekdays and by 4 p.m. on Friday. No weekend processing.

CPT Code Information

T- and B-Cell Quantitation by Flow Cytometry

86355-B cells, total count

86357-Natural killer (NK) cells, total count

86359-T cells, total count

86360-Absolute CD4/CD8 count with ratio

 

B-Cell Phenotyping Screen for Immunodeficiency and Immune Competence Assessment, Blood

86356 x7 - Mononuclear cell antigen, quantitative

 

Common Variable Immunodeficiency Confirmation Flow Panel

88184-Flow cytometry, first marker (if appropriate)

88185 x 2-Flow cytometry, each additional marker (if appropriate)

LOINC Code Information

Test ID Test Order Name Order LOINC Value
IABCS Immune Assessment B Cell Subsets, B 90416-9

 

Result ID Test Result Name Result LOINC Value
30296 CD19+ % of total Lymphocytes 8117-4
3321 CD45 Total Lymph Count 27071-0
3316 % CD3 (T Cells) 8124-0
29094 CD20+ % of total Lymphocytes 8119-0
3322 CD3 (T Cells) 8122-4
30298 CD27+ % of CD19+ B cells 89358-6
3319 % CD4 (T Cells) 8123-2
30300 CD27+ IgM+ IgD+ % of CD19+ B cells 89352-9
3325 CD4 (T Cells) 24467-3
30302 CD27+ IgM- IgD- % of CD19+ B cells 89350-3
3320 % CD8 (T Cells) 8101-8
30304 CD27+ IgM+ IgD- % of CD19+ B cells 89348-7
3326 CD8 (T Cells) 14135-8
30306 IgM+ % of CD19+ B cells 89346-1
3318 % CD19 (B Cells) 8117-4
30308 CD38+ IgM- % of CD19+ B cells 89344-6
30310 CD38+ IgM+ % of CD19+ B cells 89341-2
3324 CD19 (B Cells) 8116-6
4054 % CD16+CD56 (NK cells) 8112-5
30312 CD21+ % of CD19+ B cells 89356-0
30314 CD21- % of CD19+ B cells 89355-2
4055 CD16+CD56 (NK cells) 20402-4
3327 4/8 Ratio 54218-3
30297 CD19+ 8116-6
29095 CD20+ 9558-8
6657 Comment 80722-2
30299 CD27+ 89353-7
30301 CD27+ IgM+ IgD+ 89351-1
30303 CD27+ IgM- IgD- 89349-5
30305 CD27+ IgM+ IgD- 89347-9
30307 IgM+ 89345-3
30309 CD38+ IgM- 89343-8
30311 CD38+ IgM+ 89357-8
30313 CD21+ 25164-5
30315 CD21- 89354-5
30316 Interpretation 80722-2

Method Description

Quantitative Lymphocyte Subsets: T, B, and NK:

The T, B, and natural killer (NK)-cell surface marker assay uses monoclonal antibodies to identify the various membrane antigens, and flow cytometry to enumerate the number of cells expressing these differentiation antigens. CD14 is used to exclude monocytes, thereby improving accuracy and enhancing the purity of the lymphocyte population. The results are reported as the percent of lymphocytes that are total T cells (CD3+), CD3+CD4+ T cells, CD3+CD8+ T cells, natural killer (CD16+56+, CD3-), and B-lymphocytes (CD19+), and the absolute number of each cell type per mL of blood. The assay is a 7-color no-wash procedure and the absolute counts are calculated from internal bead standards. In addition, the total lymphocyte count and the CD4:CD8 ratio are reported.(Hoffman RA, Kung PC, Hansen WP, Goedstien G: Simple and rapid measurement of human T lymphocytes and their subclasses in peripheral blood. Proc Natl Acad Sci USA, 1980:77:4914-4917; US Department of Health and Human Services: Guidelines for performance of CD4+ T-cell determinations in persons with human immunodeficiency virus infection. MMWR Morb Mortal Wkly Rep 1997;46 no. RR-2 pp 1-29).

 

Immune Assessment B Cell Subsets, Blood:

Peripheral blood mononuclear cells (PBMC) are isolated from whole blood using a Ficoll gradient and used in the staining protocol. The assay involves a multicolor 5-tube panel for the following antibodies: CD45, CD19, CD20, CD27, IgD, IgM, CD38, and CD21. After the staining with specific antibody, the cells are washed and fixed with paraformaldehyde and then analyzed by flow cytometry on a Becton Dickinson FACS Canto instrument. The cell-surface expression is denoted as the percent of CD19+ B cells expressing each of the specific markers. CD19+ and CD20+ B cells are expressed as a percent of the total lymphocytes (CD45+). The absolute counts for the B-cell subsets are derived from flow cytometry analysis of whole blood using monoclonal antibodies to identify CD45, CD3, CD4, CD8, CD19, and CD16+CD56+. CD14 is used to exclude monocytes, thereby improving accuracy and enhancing the purity of the lymphocyte population. The assay is a 7-color, lyse-no wash procedure and the absolute counts are calculated from internal bead standards. The absolute lymphocyte count per microliter is used to calculate the absolute counts of the various B-cell subsets in this assay (Unpublished Mayo method).

 

Common Variable Immunodeficiency Confirmation Flow Panel:

Peripheral blood mononuclear cells are isolated and stained with CD19, transmembrane activator and calcium modulator and cyclophilin ligand (CAML) interactor (TACI), and B-cell-activating factor receptor (BAFF-R), each conjugated to a fluorochrome. After the staining with specific antibody, the cells are washed, fixed with paraformaldehyde, and then analyzed by flow cytometry on a Becton Dickinson FACS Canto instrument. The cell-surface expression is denoted as the percent of CD19+ B cells expressing TACI and BAFF-R.(Unpublished Mayo method)

Reject Due To

Gross hemolysis Reject
Gross lipemia Reject

Method Name

TBBS: Flow Cytometry

IABC: Fluorescent Flow Cytometry

Test Classification

See Individual Test IDs

Secondary ID

88800