Cytomegalovirus Serological Reagents

Cytomegalovirus serological reagents are devices that consist of antigens and antisera used in serological tests to identify antibodies to cytomegalovirus in serum.

The identification aids in the diagnosis of diseases caused by cytomegaloviruses (principally cytomegalic inclusion disease) and provides epidemiological information on these diseases. Cytomegalic inclusion disease is a generalized infection of infants and is caused by intrauterine or early postnatal infection with the virus. The disease may cause severe congenital abnormalities, such as microcephaly (abnormal smallness of the head), motor disability, and mental retardation. Cytomegalovirus infection has also been associated with acquired haemolytic anaemia, acute and chronic hepatitis, and an infectious mononucleosis-like syndrome.

Device Description: It is an indirect immunofluorescence test that allows detection of Human Cytomegalovirus antigen in leukocytes from peripheral blood. The test uses a monoclonal antibody pool (1C3, AYM-1) which recognizes the 65-68kDa lower matrix structural phosphoprotein (pp) (protein kinase, pp65, present in the nucleus of cells. The antibody pool (blended antibodies) recognizes two epitopes on the protein.

Intended Use: Leukocytes are prepared from whole blood by dextran sedimentation and centrifugation; slides are prepared, fixed in formalin and permeabilized on detergent (NP40). Staining is accomplished with primary murine monoclonal antibodies to the pp65 antigen and F(Ab¹) 2 fluorescein-conjugated anti-mouse immunoglobulin secondary antibody. The slides are read using a fluorescence microscope.

Intended Use: Assay is a chemiluminescent microparticle immunoassay (CMIA) used for the qualitative detection of igg antibodies to cytomegalovirus in human serum, serum separator, and plasma tubes (lithium heparin, lithium heparin separator, and tripotassium EDTA)

Device Description:  The kit consists of

• Microparticles – CMV virus lysate (strain AD169) coated microparticles in TRIS buffered saline with protein (bovine). Minimum concentration: 0.08% solids. Preservatives: ProClin 300 and antimicrobial agents.
• Conjugate – Murine anti-human IgG acridinium-labeled conjugate in MES buffer with protein (bovine). Minimum concentration: 44 ng/mL. Preservatives: sodium azide and antimicrobial agents.
• Assay Diluent – Calf serum and MES buffer with protein (bovine).

Intended Use: Identification Kit, is intended for use in the qualitative detection and identification of human cytomegalovirus (CMV) immediate early antigen (IEA) in cell cultures by immunofluorescence using fluoresceinated monoclonal antibodies (MAbs)

Device Description: Two murine-derived monoclonal antibodies (MAbs) are used in the Diagnostic Hybrids, Inc. (DHI) device, D3 DFA Cytomegalovirus Immediate Early Antigen Identification Kit (CMV-IEA ID Kit), and are directed against CMV immediate early antigen (pp 72). The MAbs used in the Kit are highly specific, with no cross-reactivity to other cultured viruses. The MAbs have been labeled by DHI using Fluorescein Isothiocyanate (FITC). The antigenemia assay is completed within two hours of blood collection, saving time and means a rapid answer for the clinician. The method consists of:

1. Direct lysis of peripheral blood erythrocytes
2. Preparation of cytospin slides
3. Fixation and permeabilization
4. Indirect immunofluorescence staining using monoclonal antibodies directed against CMV pp65 protein
5. Reading and evaluation of results

Kit Components:

• Reagent, 1 0-mL. One dropper bottle containing a mixture of two murine MAbs directed against the immediate early antigen (pp 72). The MAbs are both IgG1 (k) isotype. The buffered, stabilized, aqueous solution contains Evans Blue as a counterstain and 0.1% sodium azide as a preservative.
• Antigen Control Slides, 5-slides. Individually packaged control slides containing wells with cell culture-derived positive and negative control cells. Each slide contains one Negative well of uninfected cells and one Positive well of infected cells. Each slide is intended to be stained only one time.
• Mounting Fluid, 7-mL. One dropper bottle of an aqueous, buffered, stabilized solution of glycerol (ph 8.2 ± 0.2) and 0.1% sodium azide.
• 40X PBS Concentrate, 25-mL. One bottle containing a 40X concentrate consisting of 4% sodium azide (0.1% sodium azide after dilution to 1X using de-mineralized water) in a phosphate buffered saline (PBS) solution.

Intended Use: Test system for the detection of antibodies (igg plus igm) to cytomegalovirus (CMV) in human serum or plasma, is intended to be used in screening of patients for serological evidence of previous infection by CMV using manual and semiautomated methods

Device Description: The device is a solid-phase red cell adherence antibody detection system. The assay consists of Capture-CMV Microtitration Wells coated with glycine-extracted and purified CMV antigen obtained from cytomegalovirus strain AD 169 grown in human foreskin (HF) fibroblast cells.  Sold separately are the adjunct reagents to capture test wells and controls:

• Capture-CMV Indicator Red Cells: a suspension of human red blood cells coated with rabbit anti-human IgG plus goat anti-human IgM molecules.
• Capture LISS: a low ionic strength solution containing glycine, bromocresol purple dye and sodium azide
• Capture-CMV Positive Control Serum (weak): Human serum containing IgG antibodies to CMV viral proteins.
• Capture-CMV Negative Control Serum: Human serum containing no antibodies to CMV The CMV assay is to be used with manual, semi-automated methods.

Intended Use: Designed to be used as an independent run control with tests for the detection of Igm antibodies to Cytomegalovirus (CMV). This control is not intended as a substitute for controls provided with test kits., control is intended to estimate laboratory testing precision and can be used to detect errors in laboratory testing procedures.

Device Description: It is manufactured from human serum or plasma containing IgM antibodies to CMV but is nonreactive for Hepatitis B Surface Antigen (HBsAg) and antibodies to Human Immunodeficiency Virus Types 1 and 2 (HIV 1 and 2), antibodies to Human T-Lymphotropic Virus Type I (HTLV I) and antibodies to Hepatitis C (HCV). This control contains stabilizers (EDTA, buffering agents), and 0.1% preservative.

• Stability Studies
• Precision/Reproducibility
• Detection studies
• Linearity/assay reportable range
• Detection limit
• Analytical specificity
• Traceability (controls, calibrators, or method)
• Retrospective Study
• Analytical Sensitivity
• Assay cut-off.
• Analytical Specificity (Cross-Reactivity)
• Interfering Substances
• Precision – Intra and Inter-assay
• Prozone effect study

The same is applicable for CE Marking under IVDR also.

Required for 510k submission and IVDR CE Marking