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3244I | LIGHT DIAGNOSTICS™ CMV pp65 Antigenemia Antibody Set, included in kit #3247i

3244I
5 mL  
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      Overview

      Replacement Information

      Key Spec Table

      Key ApplicationsFormatDetection Methods
      IFPurifiedFluorescent
      Description
      Catalogue Number3244I
      Brand Family Chemicon®
      Trade Name
      • LIGHT DIAGNOSTICS
      • Chemicon
      DescriptionLIGHT DIAGNOSTICS™ CMV pp65 Antigenemia Antibody Set, included in kit #3247i
      OverviewINTENDED USE:

      The Light Diagnostics Cytomegalovirus (CMV) pp65 Antigenemia indirect immunofluorescence assay (IFA) is intended for the qualitative detection and identification of lower matrix protein pp65 of CMV in isolated peripheral blood leukocytes.

      TEST PRINCIPLE:

      The Light Diagnostics CMV pp65 Antigenemia assay utilizes an indirect immunofluorescence technique for identifying the lower matrix protein pp65 of human CMV in cytospin preparations of peripheral blood leukocytes. The blend of monoclonal antibodies provided will bind to CMV pp65 antigen present in formalin fixed leukocytes. Unbound monoclonal antibody is removed by washing with phosphate buffered saline (PBS). Fluorescein isothiocyanate (FITC) conjugated antibody will bind to the antigen-antibody complex. Unbound conjugate is removed by washing with PBS. FITC exhibits an apple green fluorescence when excited by ultraviolet light allowing visualization of the complex by fluorescence microscopy. Nuclear fluorescence indicates a positive specimen. Uninfected cells stain dull red due to the presence of Evans blue in the FITC conjugated antibody reagent.

      SUMMARY AND EXPLANATION:

      CMV is a member of the family Herpesviridae. Synthesis of viral DNA and assembly of capsids occur in the nucleus where infective progeny are released by budding through the nuclear envelope. Also characteristic of herpes viruses, CMV undergoes periods of latency and reactivation.

      CMV is species specific but has been isolated from many animal species. The first human CMV was isolated in 1956 from embryonic fibroblasts of adenoidal tissue. The terminal morphology of CMV infected permissive cells is that of a large cell with a prominent intranuclear inclusion (Cowdry Type A or "owl eye" cell). Such cells were identified in tissues of fatally infected infants which gave rise to the name "cytomegalic inclusion disease (CID)".

      Humans are believed to be the only reservoir of human CMV and postnatal infections are acquired by close contact with individuals shedding virus. Transmission may be through saliva, urine, cervical and vaginal secretions, semen, breast milk, tears, feces, and blood. Infection rates vary with geographic location and socioeconomic conditions.

      CMV infection has been detected in newborn infants and is the most commonly identified cause of congenital infection. Those infants that develop symptoms may exhibit severe disease with jaundice, hepatosplenomegaly, petechiae, and central nervous system abnormalities. The risk of infection is probably the same throughout pregnancy; CID occurs most often in fetuses infected during the first half of gestation. CMV may be transmitted to about 50% of the fetuses after primary maternal infection and about 10% of these will be clinically affected. Many congenitally infected infants appear normal at birth but subsequently develop neurologic sequelae. The route of transmission of CMV from mother to fetus has not been well elucidated. It is possible that the spread is hematogenous through cord blood or placental tissue and amniotic cells.

      During the past decade an increasing population of immunosuppressed individuals has resulted in a resurgence of CMV as a major pathogen. Induced immunosuppression has occurred more frequently via chemotherapy and transplant regimens. CMV infection is common in patients receiving renal, bone marrow, heart, lung, and liver transplants. The spread of acquired immunodeficiency syndrome (AIDS) is also related to the CMV resurgence. CMV is the most common viral opportunistic infection in AIDS patients and may be a cofactor in the pathogenesis of the AIDS virus. CMV infection in AIDS has been implicated in pneumonitis, colitis, retinitis, and dementia.

      Detection of CMV in blood leukocytes is closely associated with the clinical manifestations of CMV disease and is useful in the diagnosis of CMV infection. Rapid diagnosis of CMV disease may prevent delay in treatment using antiviral drugs such as ganciclovir and foscarnet. The

      Light Diagnostics CMV pp65 Antigenemia assay is a rapid, sensitive method for detection of CMV in isolated leukocytes.
      Materials Required but Not DeliveredReagent Preparation Required:

      · Dextran Separation Solution - 6% Dextran (MW 60,000 to 90,000) in PBS

      OR

      · Erythrocyte Lysis Buffer - 8.26 g ammonium chloride, 1 g potassium bicarbonate, 0.0037 g EDTA, QS to 1L with deionized water

      · Fixative solution - 50 mL formalin, 20 g sucrose, QS to 1L with PBS (pH 7.2 - 7.6)

      · Permeabilization Solution - 0.5% Nonidet P-40 (alternatively, IGEPAL CA-630), 10% sucrose, 1% fetal bovine serum, 0.01% sodium azide in PBS

      · Wash Solution - 1.0% fetal bovine serum in PBS and 0.1% sodium azide

      Equipment and Reagents:

      · Cytocentrifuge (Shandon Lipshaw, cytospin 3) and cytocentrifuge slides

      · Laboratory centrifuge

      · Fluorescence microscope with appropriate filter combination for FITC (excitation = 490 nm, emission = 515 nm) with 200x, 400x magnification (dry objective)

      · Hemocytometer or coulter counter

      · No. 1 microscope slide coverslips

      · Sodium hypochlorite solution, 0.05% (1:100 dilution of household bleach)

      · Humid Chamber

      · Incubator (37oC)

      · Phosphate-buffered saline (PBS)

      · Coplin staining jars

      · Deionized (DI) water

      · Hank's Balanced Salt Solution (HBSS)
      References
      Product Information
      Components
      • CMV pp65 Monoclonal Antibody - (Catalog No. 5097i) One dropper vial containing 5 mL of a blend of monoclonal antibodies against CMV pp65 antigen, protein stabilizer, 0.05% Tween 20, and less than 0.1% sodium azide.
      • Anti Mouse IgG / FITC Conjugate - (Catalog No. 5024) One dropper vial containing 10 mL of FITC labeled anti-mouse IgG antibody, 0.02% Evans blue, protein stabilizer and less than 0.1% sodium azide.
      • Mounting Fluid - (Catalog No. 5013) One dropper vial containing 10 mL of Tris buffered glycerin, a fluorescence enhancer and less than 0.1% sodium azide.
      Detection methodFluorescent
      FormatPurified
      Applications
      Key Applications
      • Immunofluorescence
      Biological Information
      Antibody TypeMonoclonal Antibody
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Availability by Geography
      • This product is not available for sale in the United States or European Union.
      Usage Statement
      • For in vitro Diagnostic Use
      • CE Mark
      Storage and Shipping Information
      Packaging Information
      Material Size5 mL
      Transport Information
      Supplemental Information
      Specifications