i.Mune™ TBNK [CE]


i.Mune™ TBNK [CE] a quantitative in vitro test to determine the percentages and absolute counts of human lymphocyte subsets in liquid venous whole blood.

i.Mune™ TBNK [CE] allows the epigenetic quantification of:

  • T lymphocytes (CD3+)
  • B lymphocytes (CD19+) (BLC)
  • Natural killer lymphocytes (CD16+CD56dim) (Natural killer cells; NKC)
  • Helper T lymphocytes (CD3+CD4+)
  • Cytotoxic T lymphocytes (CD3+CD8+).

Contents:

i.Mune™ TBNK [CE] is comprised of the three (3) separate kits:

  • i.Mune™ Prep (for preparation of a total of 48 samples)
  • i.Mune™ TBNK Amp (for PCR-amplification of a total of 48 samples)
  • i.Mune™ Check (controls and standards or a total of 12 DNA prep runs)

In addition, we'll provide a Data Analysis Tool (MS-Excel based)



Sample requirements:

  • 40 µl of liquid venous whole blood collected in K2EDTA blood collection tube.
  • The blood sample may be stored at room temperature (15°C to 30°C) for up to 48 h

Quantification of T-/B- and NK lymphocytes can be useful for*:

  • Follow-up and diagnostic evaluation of primary immunodeficiency 1),2)
  • Monitoring of HIV-positive patients 1),3)
  • Immune monitoring following immunosuppressive therapy for transplantation, autoimmunity and other immunological conditions4)
  • Assessment of immune reconstitution post hematopoietic stem cell transplantation5)
  • Early screening of gross quantitative anomalies in lymphocyte subsets in infections and malignancies
  • Absolute quantification of circulating B cells for diagnosis of chronic lymphocytic leukemia (CLL) patients6)

Please inquire to obtain a customized proposal:

*The above clinical applications have been established using technologies currently being employed in clinical laboratory routine (e.g. flow cytometry). 

Performance Characteristics:

Performance characteristics of i.Mune™ TBNK [CE] were determined according to CLSI guidelines.

Repeatability (single-site) CV < 12,5%                                     
Within Laboratory Precision (single-site)

CV < 25%

Reproducibility (multi-site)

CV < 25%

Within Laboratory Precision (multi-site)

CV < 25%

Linearity

10 - 40 µl venous whole blood

Limit of Quantification

< 100 copies

Reliability

99%

Interference

No interference with bilirubin (0,4 mg/ml),
hemoglobin (10 mg/ml),
triglycerides (15 mg/ml), albumin (150 mg/ml)

Clinical Performance:

Clinical performance of i.Mune™ TBNK [CE] was evaluated according to DIN EN 13612. For this 113 liquid venous whole blood samples from self-declared healthy donors were prospectively collected in K2-EDTA blood collection tubes. Epigenetic data of 112 test samples analysed with i.Mune™ TBNK [CE] were compared to flow cytometry data using the Spearman correlation coefficient (Spearman r) for each assay (% and cells/μl).

Assay Spearman r p value
 CD3 cells/µl 0.82 <0.0001
CD4 cells/µl 0.70  <0.0001
CD8 cells/µl 0.55  <0.0001
BLC cells/µl 0.79  <0.0001
NKC cells/µl 0.70  <0.0001
Leukocytes/µl 0.70  <0.0001
     
CD3 % of Leukocytes 0.85 <0.0001
CD4 % of Leukocytes 0.76 <0.0001
CD8 % of Leukocytes 0.53 <0.0001
BLC % of Leukocytes 0.86 <0.0001
NKC % of Leukocytes 0.78 <0.0001

Comparison i.Mune™ TBNK [CE] with flow cytometry

(n=112 self declared healthy adult donors)

 

r = 0.90

r = 0.96


References

  1. Baron U. et al. Epigenetic immune cell counting in human blood samples for immunodiagnostics. Sci Transl Med. 2018 Aug 1; 10 (452)
  2. Aluri J, et al. Clinical, Immunological, and Molecular Findings in 57 Patients With Severe Combined Immunodeficiency (SCID) From India. Front Immunol. 2019 Feb 4;10:23
  3. Ford N, et al. The evolving role of CD4 cell counts in HIV care. Curr Opin HIV AIDS. 2017 Mar;12(2):123-128
  4. Omana-Zapata I, et al. Accurate and reproducible enumeration of T-, B-, and NK lymphocytes using the BD FACSLyric 10-color system: A multisite clinical evaluation. PLoS One. 2019 Jan 28;14(1):e0211207
  5. Riley RS. Laboratory evaluation of the cellular immune system. In: McPherson RA, Pincus MR, eds. Henry's Clinical Diagnosis and Management by Laboratory Methods. 23rd ed. St Louis, MO: Elsevier; 2017:chap 45
  6. Hallek M, et al. International Workshop on Chronic Lymphocytic Leukemia. Guidelines for the diagnosis and treatment of chronic lymphocytic leukemia: a report from the International Workshop on Chronic Lymphocytic Leukemia updating the National Cancer Institute-Working Group 1996 guidelines. Blood. 2008 Jun 15;111(12):5446-56