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Test ID: Lymphocyte Subset Analysis
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Lymphocyte Subset Analysis
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Useful For
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- Serial monitoring of CD4 T-cell count in HIV-positive patients.
- Immune monitoring following immunosuppressive therapy for transplantation, autoimmunity, and other immunological conditions where such treatment is utilized.
- Assessment of immune reconstitution posthematopoietic cell transplantation.
- Early screening of gross quantitative anomalies in lymphocyte subsets in infection or malignancies.
- Absolute quantitation of circulating B cells for diagnosis of chronic lymphocytic leukemia patients.
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Method name and description
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Method Name: Flowcytometry
When whole blood is added to the reagent (BDMultitest™ 6-color TBNK reagent with BD Trucount™ tubes) the fluorochrome-labeled antibodies in the reagent bind specifically to leucocyte surface antigens. During acquisition, the cells travel past the laser beam and scatter the laser light. The stained cells fluoresce. These scatter and fluorescence signals, detected by the instrument, provide information about the cell’s size, internal complexity, and relative fluorescence intensity.
BD Multitest reagents employ fluorescence triggering, allowing direct fluorescence gating of the lymphocyte population to reduce contamination of unlysed or nucleated red blood cells in the gate.
When BD Trucount tubes are used, the lyophilized pellet in the tube dissolves, releasing a known number of fluorescent beads. During analysis, the absolute number (cells/μL) of positive cells in the sample can be determined by comparing cellular events to bead events.
Test is done by stain - lysis technique and analyzed by Flow cytometer.
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Clinical information
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Lymphocyte Subset Analysis is used to identify and determine the percentages and absolute counts of T cells (CD4,CD8),B cells (CD19), and Natural Killer (CD16+CD56) cells in peripheral blood. Each of these lymphocyte subpopulations have distinct effector and regulatory functions and are maintained in homeostasis under normal physiological conditions. The CD3 antigen is a pan-T cell marker, and T cells can be further divided into 2 broad categories, based on the expression of CD4 or CD8 coreceptors. B cells can be identified by expression of CD19, while NK cells are typically identified by the coexpression of CD16 and CD56.
The lymphocytes subset test is used for monitoring of CD4 T-cell count in HIV-positive patients, immune monitoring following immunosuppressive therapy for transplantation, autoimmunity, and other immunological conditions. It is uses as well for assessment of immune reconstitution posthematopoietic cell transplantatio, early screening of gross quantitative anomalies in lymphocyte subsets in infection or malignancies and for quantitation of absolute count of circulating B cells for diagnosis of chronic lymphocytic leukemia patients and monoclonal b-lymphocytosis.
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Aliases
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Cell Phenotypic Analysis
Lymphocyte Subset Analysis Test
T cell Subset
T,B,NK cell count
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Specimen type / Specimen volume / Specimen container
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Specimen type: A fresh whole blood sample.
Container Tube: EDTA Tube.
Specimen Volume: 3 mL,Whole Blood EDTA, For pediatric 500ul (EDTA microtainer).
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Collection instructions / Special Precautions / Timing of collection
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- Specimens should be processed within 24 hours of sampling.
- Transport Immediately after collection in room temperature.
- Routine: Sunday thruThursday.
- Last samples on Thursday should be collected before 11:00 am.
- Samples Collected on the weekends will be rejected.
- Date and time of draw is required.
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Relevant clinical information to be provided
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Sufficient clinical information relevant to the requested test should be provided.
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Storage and transport instructions
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- The specimen should be sent to the laboratory as soon as possbile after collection.
- The sample should be stored at room temperature within 24 hours of blood collection.
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Specimen Rejection Criteria
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Insufficient quantity of blood.
Clotted.
Wrong collection tube or anticoagulant.
Outdated specimens .
Improper sample storage.
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Biological reference intervals and clinical decision values
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| Cell Phenotypic Analysis (Lymphocyte Subset Analysis Test) Reference Range |
| Age |
T-Cell Surface Antigen |
B-Cell and NK Surface Antigen |
| CD3+ |
CD3+/CD4+ |
CD3+/CD8+ |
CD19+ |
CD3-/ CD16+/ CD56+ |
| %Pos. |
Cells/ul |
%Pos. |
Cells/ul |
%Pos. |
Cells/ul |
%Pos. |
Cells/ul |
%Pos. |
Cells/ul |
| Neonate |
28-76 |
600-5000 |
17-52 |
400-3500 |
10-41 |
200-1900 |
5-22 |
40-1100 |
6-58 |
100-1900 |
| 1 wk- 2 months |
60-85 |
2300-7000 |
41-68 |
1700-5300 |
9-23 |
400-1700 |
4-26 |
600-1900 |
3-23 |
200-1400 |
| 2- 5 months |
48-75 |
2300-6500 |
33-58 |
1500-5000 |
11-25 |
500-1600 |
14-39 |
600-3000 |
2-14 |
100-1300 |
| 5-9 months |
50-77 |
2400-6900 |
33-58 |
1400-5100 |
13-26 |
600-2200 |
13-35 |
700-2500 |
2-13 |
100-1000 |
| 9-15 months |
54-76 |
1600-6700 |
31-54 |
1000-4600 |
12-28 |
400-2100 |
15-39 |
600-2700 |
3-17 |
200-1200 |
| 15-24 months |
39-73 |
1400-8000 |
25-50 |
900-5500 |
11-32 |
400-2300 |
17-41 |
600-3100 |
3-16 |
100-1400 |
| 2-5 years |
43-76 |
900-4500 |
23-48 |
500-2400 |
14-33 |
300-1600 |
14-44 |
200-2100 |
4-23 |
100-1000 |
| 5-10 years |
55-78 |
700-4200 |
27-53 |
300-2000 |
19-34 |
300-1800 |
10-31 |
200-1600 |
4-26 |
90-900 |
| 10-18 years |
52-78 |
800-3500 |
25-48 |
400-2100 |
9-35 |
200-1200 |
8-24 |
200-600 |
6-27 |
70-1200 |
| Adults |
57.52-83.11 |
856-2669 |
31.45-62.38 |
491-1734 |
9.55-38.32 |
162-1074 |
5.89-24.21 |
73-562 |
5.17-30.36 |
108-680 |
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Turnaround time / Days and times test performed / Specimen retention time
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Turnaround time: 24 working Hours.
Days and times test performed: Sunday through Thursday, Last samples on Thursday should be collected before 11:00 am
For serial monitoring, we recommend that specimen draws be performed at the same time of day.
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