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3430 | LIGHT DIAGNOSTICS™ Varicella-Zoster Virus DFA Kit, ~125 tests

3430
5 mL  
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      Overview

      Replacement Information

      Key Spec Table

      Key ApplicationsFormatHostDetection Methods
      IFFITCMFluorescent
      Description
      Catalogue Number3430
      Brand Family Chemicon®
      Trade Name
      • LIGHT DIAGNOSTICS
      • Chemicon
      DescriptionLIGHT DIAGNOSTICS™ Varicella-Zoster Virus DFA Kit, ~125 tests
      OverviewThe Light Diagnostics Varicella-zoster Virus (VZV) Direct Immuno-fluorescence Assay (DFA) is intended for the qualitative detection and identification of GPI and immediate early antigen of VZV from vesicular lesions. The kit is intended for use in culture confirmation with standard tube cultures and shell vials and is presumptive in the detection and identification of VZV from direct specimens.

      For in vitro diagnostic use.

      Test Principle:

      Light Diagnostics VZV Direct Immunofluorescence Assay utilizes a direct immunofluorescence technique for identifying virus in direct specimens and infected tissue cultures. The FITC labeled monoclonal antibody reagent provided will bind to VZV antigen (glycoprotein gp I and the immediate early antigen) if present in the sample. Unbound FITC labeled monoclonal antibody is removed by rinsing with Phosphate Buffered Saline (PBS). FITC exhibits an apple green fluorescence when illuminated by ultraviolet light allowing visualization of the antigen antibody complex by fluorescence microscopy. Cell fluorescence indicates a positive specimen. Uninfected cells stain a dull red due to the presence of Evans Blue in the FITC-labeled monoclonal antibody.

      Summary and Explanation:

      Varicella-zoster virus, a member of the family Herpesviridae, causes two different clinical syndromes: chickenpox (varicella) and shingles (zoster). Varicella occurs upon primary infection whereas reactivation of latent virus, a characteristic of herpes viruses, causes zoster. Morphologically, VZV shares the structure of the Herpesviridae family with a linear double stranded DNA. The virion consists of an inner capsid surrounded by a membrane envelope with a diameter of 150-200 nm.

      Varicella occurs most frequently in children. The most notable clinical feature is a generalized vesicular rash which may be accompanied by fever. In contrast, zoster generally occurs in adults. This may be a reactivation of latent virus or persistent infection where some VZV is constantly being produced. Zoster is characterized by a painful eruption of vesicular lesions with inflammation of the associated sensory nerve ganglion.

      Complications associated with varicella occur primarily in neonates, adults and immunocompromised individuals resulting in increased morbidity and mortality. Varicella pneumonia is the most serious complication with mortality ranging between 10 and 40%1. Pregnant women are at greater risk for varicella pneumonia than other adults. The fatality rate from neonatal varicella may exceed 30% when maternal varicella occurs between 5 days before and 2 days after birth2,3.

      The most common complication of zoster is postherpetic neuralgia. Delayed healing may result in secondary bacterial infection or gangrene in both zoster and varicella. Complications and severity of zoster are significantly increased in immunosuppressed patients.6,7

      Antiviral agents are now available for the treatment of VZV infection. Effective antiviral therapy, especially in immunosuppressed patients, is dependent upon specific diagnosis differentiating VZV from herpes simplex virus4. Accurate laboratory diagnosis of the less common manifestations of VZV infection, such as varicella pneumonia and encephalitic complications, is also important for the institution of treatment.

      Direct detection by immunofluorescence and culture isolation with confirmation provide definitive diagnosis of VZV infection5. Light Diagnostics Varicella-zoster Virus Direct Immunofluorescence Assay is a rapid, accurate method for identification of VZV in direct specimens, centrifugation enhanced shell vials and standard tube cultures.
      Alternate Names
      • VZV
      Materials Required but Not Delivered· Acetone, reagent grade; stored in glass

      · Distilled water

      · Positive controls, for culture isolation procedures (reference VZV strains available from ATCC, Rockville, MD: Webster VR-916, Ellen VR-586 and Oka VR-795)

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

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

      · Tissue culture media (RPMI or Eagle's Minimum Essential Medium with Precolostral Bovine Serum and antibiotics or equivalent)

      · Viral transport medium which is non-inhibitory to VZV (Hanks balanced salt solution with antibiotics or equivalent)

      · 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 100 watt mercury or halogen lamp illumination and the appropriate filter combination for FITC (excitation peak 490 nm, emission peak 520 nm) with 100x, 200x, 400x magnification (dry objective)

      · Forceps

      · Humid chamber

      · Incubator, 37 + 1°C

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

      · Ultrasonic water bath

      · Vortex mixer or sonicator
      References
      Product Information
      Components
      • VZV FITC Labeled Monoclonal Antibody - (Catalog No. 5094). One (1) 5 ml dropper vial containing a blend of FITC conjugated, whole IgG purified monoclonal antibodies against gp I and immediate early antigen of VZV, protein stabilizer, 0.005% Evans Blue, 0.05% Tween 20 and 0.1% Sodium Azide (preservative).
      • VZV Control Slides - (Catalog No. 5088). Two (2) slides containing one well of VZV (clinical isolate) infected human foreskin fibroblasts (positive) and one well of non-infected human foreskin fibroblasts (negative).
      • Phosphate Buffered Saline (PBS) - (Catalog No. 5087). One (1) packet of Phosphate Buffered Saline salts.
      • Tween 20 / Sodium Azide Solution (100X) - (Catalog No. 5037). One (1) 10 ml vial containing Tween 20 / Sodium Azide concentrate.
      • Mounting Fluid - (Catalog No. 5013). One (1) 10 ml dropper vial containing Tris-buffered Glycerin, a fluorescence enhancer and 0.1% Sodium Azide (preservative).
      Detection methodFluorescent
      FormatFITC
      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
      • For in vitro Diagnostic Use
      • CE Mark
      Storage and Shipping Information
      Storage Conditions· When stored at 2-8°C, the VZV 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:

      * For in vitro diagnostic use.

      * Sodium Azide (present in the conjugate, PBS and mounting fluid) 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.

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

      * Do not mix or substitute reagents from different kit lots.

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

      * Handle all specimens and materials as potentially infectious materials. Decontaminate 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-VZV FITC labeled monoclonal antibody) as it is a potential carcinogen. If skin contact occurs, flush with large volumes of water.

      * Do not mouth pipette reagents.
      Packaging Information
      Material Size5 mL
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      SDS

      Title

      Safety Data Sheet (SDS) 

      References

      Reference overviewPub Med ID
      DNA double-strand breaks in heterochromatin elicit fast repair protein recruitment, histone H2AX phosphorylation and relocation to euchromatin.
      Burkhard Jakob,Jörn Splinter,Sandro Conrad,Kay-Obbe Voss,Daniele Zink,Marco Durante,Markus Löbrich,Gisela Taucher-Scholz
      Nucleic acids research 39 2011

      Show Abstract Full Text Article
      21511815 21511815

      User Guides

      Title
      LIGHT DIAGNOSTICS TM KIT DFA DE VARICELLA-ZOSTER VIRUS multi languages
      LIGHT DIAGNOSTICSTM VARICELLA-ZOSTER VIRUS DFA KIT