CD4 easy count kit – dry

Product Name CD4 easy count kit - dry IVD

Code No. 05-8401-d

Content Packing contains reagents for 100 tests:

• 100 sample tubes (green cap) with lyophilized CD4 mAb PE (MEM-241,
PE-conjugated monoclonal antibody to human CD4)
• 100 sample tubes (white cap) with each 820 µl CD4 buffer

General Information: Patients with AIDS exhibit T-cell
lymphopenia, a loss of CD4+ lymphocytes and a relative
increase in the CD8+ subtype and in the CD3+CD4-CD8-
subtype. A precise enumeration of CD4+ T-cells is
necessary for a reliable and controlled antiretroviral therapy
and patient monitoring.
Specificity and Antigen Distribution: The mouse
monoclonal antibody MEM-241 recognizes the human CD4
antigen, a transmembrane glycoprotein (55 kDa) of the
immunoglobulin supergene family, present on a subset of
T-lymphocytes (“helper/inducer” T-cells) and also
expressed at a lower level on monocytes, tissue
macrophages and granulocytes. Approximately 20-60% of
human peripheral blood mononuclear cells as well as a
subpopulation of monocytes but with a weaker signal are
stained. The antibody has been studied at the 8th
International Workshop on Human Cell Differentiation
Molecules HCDM (former HLDA VIII), May 2006, Québec,
Canada. CD4 is the primary cellular receptor for the human
immunodeficiency virus (HIV).
Clone: MEM-241, Isotype: Mouse monoclonal IgG1
Applications:
• Identification and enumeration of the CD4+ helper/inducer
T-lymphocytes subset
• Studies of HIV infection. Patients infected with HIV loose
CD4+ helper/inducer lymphocytes
• Characterization of subtypes of T-cell leukemias and
lymphomas
• For in vitro diagnostic use
Method:
• Add 20 µl whole blood (EDTA as anticoagulant) to a
Partec sample tube (green cap) with lyophilized CD4
mAb PE. Add blood to the bottom of the tube
• Mix gently and incubate for 15 minutes at room
temperature protected from light
• After 5 minutes incubation time mix again gently
• Pour 820 µl CD4 buffer out of sample tube (white cap) to
the stained blood sample and shake gently
• Analyze blood sample on a Partec device
Flow Cytometric Analysis: CD4-PE fluorescence can be
analysed on a Partec Flow Cytometer with an excitation light
source of 488 nm or 532 nm (blue or green solid state laser).
To count CD4+ T-cells transfer the test tube with 840 µl of the
ready prepared blood sample (see Method) to your Partec
Flow Cytometer and carry out a count analysis. For detailed
information on the handling and calibration of the instrument
please refer to the operating manuals and application notes.
The blood preparation allows one count procedure on the
same sample. Following the above procedure the dilution
factor is 42. In the CyFlow® Counter and CyFlow® SL_3 this
dilution factor is already set or can be directly set by the
instrument operator in the instrument software. Counting
results will be displayed automatically as CD4+ T-cells per µl
whole blood.
Recommended and Required Materials:
• Partec Flow Cytometry instrument (e.g. CyFlow® Counter
or CyFlow® SL_3)
• Partec test tubes (Code No. 04-2000)
• Micropipettes and pipette tips (e.g. Eppendorf, Code No.
3112 000.029 and 3111 000.0165)
• Powder-free latex gloves (e.g. Safeskin, Code No. 545-950-
06)
• Venous Blood Collection System with EDTA as
anticoagulant (e.g. Greiner Bio-One: Vacuette® EDTA
Tubes, K3E/EDTA K3, 3 ml, Code No. 454217, Vacuette®
Blood Collection Needles 38 x 0.8 mm, Code No. 450076,
Vacuette® Tube Holders, Code No. 450201)
Handling and Storage: CD4 mAb PE is supplied as
lyophilisate. The amount of lyophilized CD4 mAb PE in each
sample tube is sufficient for labelling 1x106 cells. Store the
sample tubes with lyophilized CD4 mAb PE at 2-35°C,
protected from light and humidity. Store the CD4 buffer
at 2-35°C. Do not freeze or expose to elevated temperatures.
The CD4 easy count kit - dry is stable until the expiration date
printed on the kit label.

For in vitro diagnostic use with Partec IVD Flow Cytometers.

Contact Information:

Partec GmbH • Am Flugplatz 13 • D-02828 Görlitz • Germany

Tel +49 3581 8746 0 • Fax +49 3581 8746 70 • E-mail: mail@partec.com

Rev. 005 Date 2009-07-24 05-8401-d CD4 easy count kit - dry
 CD4 easy count kit – dry – Page 2
Example of a direct CD4 counting result using the CD4
easy count kit – printout from a Partec CyFlow® Counter. As
depicted in the histogram, the CD4+ T-cells (prominent peak on
the right) can be clearly separated from the CD4+ monocytes
(left peak of weaker fluorescence intensity). The absolute
concentration of CD4+ T-cells is displayed directly as number
of cells/µl whole blood
Example of a direct CD4 counting result using the CD4
easy count kit – display of a Partec CyFlow® SL_3. The FL2
CD4 PE histogram plot shows the precise separation of the
CD4+ T-cell peak (cell population in the RN1 range) from
other cell distributions.
Literature:
Cassens, U. et al.: Simplified volumetric flow cytometry allows
feasible and accurate determination of CD4 T lymphocytes in
immunodeficient patients worldwide”, Antiviral Therapy 9:
395-405 (2004)
Göhde, W. et al.: A New Protocol to Follow-up Immune
Impairment in HIV/AIDS-Children, Scientific Presentation -
Abstract No. MoPeLB151C02 at the 3rd IAS Conference on
HIV Pathogenesis and Treatment in Rio de Janeiro, July 2005
Greve, B. et al.: ”A New No-Lyse, No-Wash Flow-Cytometric
Method for the Determination of CD4 T Cells in Blood
Samples”, Transfus Med Hemoth (2003), 30: 8-13
Imade, G. et al.: “Comparison of a New, Affordable Flow
Cytometric Method and the Manual Magnetic Bead
Technique for CD4 T-Lymphocyte Counting in a Northern
Nigerian Setting”, Clinical and Diagnostic Laboratory
Immunology, Jan. 2005, p. 224-227
Dieye, T.N. et al.: Absolute CD4 T-Cell Counting in Resource-
Poor Settings: Direct Volumetric Measurements Versus Bead-
Based Clinical Flow Cytometry Instruments, Journal of
Acquired Immune Deficiency Syndromes, Volume 39,
Number 1, May 1 2005
Fryland, M. et al.: The PARTEC CyFlow counter for CD4+ T-
cell counting produces high quality results and is robust when
evaluated under routine field conditions in Malawi, Scientific
Oral Presentation - Abstract No. TuOrB1149 at the XV.
International AIDS Conference in Bangkok, July 2004

Warnings and Precautions: Reagents contain 0.09%

sodium azide that under acidic conditions yields hydrazoic
acid, an explosive compound. Sodium azide may react with
lead and copper plumbing to form potentially explosive metal
azides. When disposing of sodium azide, flush plumbing with
a large volume of water to prevent build-up. Very toxic if
swallowed. After contact with skin, wash immediately with
plenty of water.
R22: Harmful if swallowed, R32: Contact with acids liberates
very toxic gas, S2: Keep out of reach of children, S13: Keep
away from food, drink and animal feeding stuffs, S36: Wear
suitable protective clothing, S46: If swallowed, seek medical
advice immediately and show this container or label

Handle all specimens and materials as potentially infectious

materials. Decontaminate with 0.05% sodium hypochlorite.
The product should be handled by trained staff only.
Reagents are not sterile. Not for in vivo use. For in vitro use
only.

For in vitro diagnostic use with Partec IVD Flow Cytometers.

Contact Information:

Partec GmbH • Am Flugplatz 13 • D-02828 Görlitz • Germany

Tel +49 3581 8746 0 • Fax +49 3581 8746 70 • E-mail: mail@partec.com

Rev. 005 Date 2009-07-24 05-8401-d CD4 easy count kit - dry