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3135 | LIGHT DIAGNOSTICS™ CMV IFA Kit, ~250 tests

3135
10 mL  
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      Key ApplicationsFormatHostDetection Methods
      IF Purified M Fluorescent
      Description
      Catalogue Number3135
      Brand Family Chemicon®
      Trade Name
      • LIGHT DIAGNOSTICS
      • Chemicon
      DescriptionLIGHT DIAGNOSTICS™ CMV IFA Kit, ~250 tests
      OverviewThe Light Diagnostics Cytomegalovirus (CMV) Immunofluorescence Assay (IFA) is intended for use in culture confirmation with standard culture tubes and shell vials in the qualitative detection and identification of immediate early antigen of human CMV in vitro.

      This product is not FDA approved for use in testing blood or plasma donors.

      Test Principle:

      Light Diagnostics Cytomegalovirus Immunofluorescence Assay utilizes an indirect immunofluorescence technique for identifying the immediate early antigen (IEA) of human cytomegalovirus in infected tissue cultures. The monoclonal antibody provided will bind to CMV immediate early antigen present in the cell. Unbound monoclonal antibody is removed by rinsing with phosphate-buffered saline (PBS). FITC (fluorescein isothiocyanate) - conjugated second antibody is then added which will bind to the antigen-antibody complex. Unbound secondary antibody is removed by rinsing with PBS. FITC exhibits an apple green fluorescence when illuminated by ultraviolet light allowing visualization of the complex by fluorescence microscopy. Fluorescence of the cell nuclei indicates a positive specimen. Non-infected cells stain a dull red due to the presence of Evans blue in the FITC-conjugated secondary antibody.

      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 and has been isolated from many animal species (1). The first human CMV was isolated in 1956 from embryonic fibroblasts of adenoidal tissue (2). 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)(3).

      Humans are believed to be the only reservoir of human CMV. Postnatal infections are acquired by close contact with individuals shedding virus through saliva, urine, cervical and vaginal secretions, semen, breast milk, tears, feces, and blood (4,5,6). Infection rates vary with geographic location and socioeconomic conditions.

      CMV infection in newborn infants is the most commonly identified cause of congenital infection. Those infants which 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 (7,8). Many congenitally infected infants appear normal at birth but may 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 (9), bone marrow (10), heart (11), lung (12), and liver transplants (13). The rapid spread of acquired immunodeficiency syndrome (AIDS) is also related to the CMV resurgence. CMV is the most common viral opportunistic infection (14,15) and has been considered as a cofactor in the pathogenesis of the AIDS virus (16). CMV infection in AIDS has been implicated in pneumonitis (17), colitis (18,19), retinitis (20), and dementia (21).

      Light Diagnostics Cytomegalovirus Immunofluorescence Assay enables pre-CPE detection of CMV in centrifugation enhanced shell vials with an early detection option at *48 hours after specimen inoculation. The kit provides sufficient reagents to process 60 shell vials or 300 culture tubes.
      Materials Required but Not Delivered· Distilled water

      · Acetone, reagent grade; stored in glass

      · Negative and positive controls (representative CMV strains available from ATCC, Rockville, MD).

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

      · Sterile shell vials with 12 mm coverslips for growth of MRC-5 or CMV permissive human fibroblasts or culture tubes with MRC-5 or CMV permissive human fibroblasts

      · Tissue culture media RPMI or Eagle's minimum essential medium (EMEM) with fetal bovine serum (FBS) and antibiotics or equivalent

      · Viral transport medium which is noninhibitory to CMV (Hanks balanced salt solution (HBSS) with antibiotics or equivalent)

      · Sterile PBS without sodium azide

      · 0.1N NaOH

      · 0.1N HCl

      · Microscope slides, non-fluorescing

      · No. 1 coverslips

      · Aspirator device with disposable sterile Pasteur pipettes

      · Centrifuge capable of 700-950 x g with biohazard buckets and adapters for shell vials

      · Fluorescence microscope with appropriate filter combination for FITC (excitation peak = 490 nm, emission peak = 520 nm) with 100x, 200x, 400x magnification (dry objective)

      · Forceps

      · Humid chamber

      · CO2 Incubator, 37+1°C

      · Syringe and needle or other implement to remove coverslip from shell vial

      · Syringe filter, 0.45 micron

      · Ultrasonic water bath

      · Vortex mixer or sonicator
      References
      Product Information
      Components
      • CMV Monoclonal Antibody - (Catalog No. 5026). One 10 ml dropper vial containing monoclonal antibody against CMV immediate early antigen, protein stabilizer and 0.1% sodium azide (preservative).
      • Anti-Mouse IgG : FITC Conjugate - (Catalog No. 5024). One 10 ml dropper vial containing FITC-conjugated anti-mouse IgG antibody, 0.02% Evans blue, protein stabilizer and 0.1% sodium azide (preservative).
      • CMV Control Slides - (Catalog No. 5027). Two slides containing one well of CMV- infected cells (positive) and one well of non-infected cells (negative).
      • -Phosphate Buffered Saline (PBS) - (Catalog No. 5087). One packet of phosphate buffered saline salts.
      • Tween 20 / Sodium Azide Solution (100X) - (Catalog No. 5037). One 10 ml vial containing Tween 20 / sodium azide concentrate.
      • Mounting Fluid - (Catalog No. 5013). One 10 ml dropper vial containing Tris-buffered glycerin, a fluorescence enhancer and 0.1% sodium azide (preservative).
      Detection methodFluorescent
      FormatPurified
      Applications
      Key Applications
      • Immunofluorescence
      Biological Information
      HostMouse
      Antibody TypeMonoclonal Antibody
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Usage Statement
      • CE Mark
      • For in vitro Diagnostic Use
      Storage and Shipping Information
      Storage ConditionsWhen stored at 2 - 8°C, the CMV kit is stable up to the expiration date printed on the kit label. Do not freeze or expose to elevated temperatures. Discard any remaining reagents after the kit expiration date.

      Warnings and Precautions:

      * Sodium azide, present in the conjugate, antibody, PBS/Tween and Mounting Fuid, can form potentially explosive metal azides with lead and copper pipes. As a precaution, flush with large amounts of water to prevent azide build-up.

      * Pooling, substitution or alteration of any reagent may cause erroneous results.

      * Do not mix reagents from different lots.

      * Do not allow the slides to dry at any time during the staining procedure.

      * Handle all specimens and materials coming in contact with them as potentially infectious materials. Disinfect with 0.05% sodium hypochlorite.

      * Acetone is extremely flammable and harmful if swallowed or inhaled. Keep away from heat, sparks or flame. Avoid breathing vapor. Use adequate ventilation.

      * Avoid contact with Evans blue (present in the anti-mouse IgG : FITC conjugate) as it is a potential carcinogen. If skin contact occurs, flush with large volumes of water.

      * Do not mouth pipette reagents.

      * Mounting Fluid contains a fluorescence enhancer that may be destructive to mucous membranes. Avoid direct skin or mucous membrane contact. If contact occurs, flush with large volumes of water.
      Packaging Information
      Material Size10 mL
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      MSDS

      Titolo

      Scheda di sicurezza (MSDS) 

      Riferimenti bibliografici

      Panoramica dei riferimenti bibliograficiCodice d'identificazione nel Pub Med
      A novel protein kinase D inhibitor attenuates early events of experimental pancreatitis in isolated rat acini.
      Edwin C Thrower,Jingzhen Yuan,Ashar Usmani,Yannan Liu,Courtney Jones,Samantha N Minervini,Martine Alexandre,Stephen J Pandol,Sushovan Guha
      American journal of physiology. Gastrointestinal and liver physiology 300 2011

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      20947701 20947701

      Manuali d'uso

      Titolo
      KIT IFA CMV
      LIGHT DIAGNOSTICS™ CMV IFA KIT Pre-CPE Culture Identification Test