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3295 | LIGHT DIAGNOSTICS™ SimulFluor® HSV/VZV, ~125 tests

3295
5 mL  
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      Overview

      Replacement Information

      Key Spec Table

      Key ApplicationsFormatHostDetection Methods
      IFSimulFluorMFluorescent
      Description
      Catalogue Number3295
      Brand Family Chemicon®
      Trade Name
      • LIGHT DIAGNOSTICS
      • SimulFluor
      • Chemicon
      DescriptionLIGHT DIAGNOSTICS™ SimulFluor® HSV/VZV, ~125 tests
      OverviewThe Light Diagnostics SimulFluor® HSV/VZV Immunofluorescence Assay is intended for the simultaneous detection and identification of herpes simplex viruses (HSV) 1 and 2 and varicella-zoster virus (VZV) from patients with vesicular, oral, genital, or skin lesions, using direct specimens and culture confirmation. Specimens found to be negative on direct specimen examination must be confirmed with culture.

      For in vitro diagnostic use

      Test Principle:

      Light Diagnostics SimulFluor® HSV/VZV Immunofluorescence Assay utilizes a single reagent for the simultaneous detection and identification of HSV and VZV. The primary component containing monoclonal antibodies, specific for both HSV 1 and 2 will bind to 155kD major capsid protein in HSV-infected cells. The secondary component containing monoclonal antibodies, specific for VZV, will bind to glycoprotein gp I and the immediate early antigen in VZV-infected cells. Unbound reagent is removed by rinsing with phosphate-buffered saline (PBS). Illumination with ultraviolet light allows visualization of the antigen-antibody complexes by fluorescence microscopy. When a FITC filter set is used, the HSV antigen-antibody complex will exhibit an apple-green fluorescence and the VZV antigen-antibody complex will fluoresce yellow-gold. Uninfected cells stain a dull red due to the presence of Evans blue in the reagent.

      Summary and Explanation :

      Herpes simplex virus (HSV) and varicella-zoster virus (VZV) are members of the Herpesviridae family. They are large, enveloped viruses, about 190 nm in diameter containing linear, double-stranded DNA. Both viruses cause a multitude of human diseases and are particularly severe in immunocompromised patients.

      There are two biologically distinct serotypes of HSV classified as type 1 (HSV-1) and type 2 (HSV-2) The serotypes are closely related with extensive sequence homologies of their DNA's. HSV is a major cause of conjunctival, respiratory, central nervous system, genital, and generalized disease (1-4). HSV infects mucocutaneous surfaces, then enters the dorsal root ganglia where further viral replication occurs, followed by a period of latency. Reactivation is accompanied by viral excretion at, or close to, the original mucocutaneous site of infection, with or without, the associated clinical signs and symptoms. Recurrent lesions are usually less severe than the primary infection.

      HSV-1 causes gingivostomatitis, intense pharyngitis, tonsillitis, and occasionally encephalitis in infants and children during their primary infections. Ocular, nasal, orolabial, and oropharyngeal lesions occur in both children and adults. Due to the ubiquity of HSV-1 and its ease of spread by aerosolized droplets, fomites, and direct contact, most adults experience HSV-1 infection during their lifetime.

      HSV-2 is more frequently associated with painful genital lesions, urethritis, and cervicitis in adults. If the virus is present in the birth canal around the time of delivery, either from primary or recurrent infection, severe generalized infection in the neonate may occur. Thus, maternal genital HSV infections pose a substantial risk to the fetus and newborn.

      Shedding of virus at the time of delivery is frequently the route of transmission from mother to neonate. Neonatal HSV infection is generally symptomatic and often fatal, with a mortality rate in untreated cases of 70%. The clinical presentation may be localized infection of the skin, eyes and mucosa, encephalitis, or disseminated disease.

      Acyclovir, famciclovir, foscarnet, and other nucleoside analogs can reduce clinical symptoms and virus shedding in oral and genital herpes, herpes encephalitis, neonatal herpes, and herpetic keratitis (5-7).

      HSV can be readily recovered from clinical specimens following culture in cells lines such as HEp2, MRC-5, A549, HEK, NCI-H292, and others. Incubation time in stationary or roller cultures ranges from 1 to 7 days with evident cytopathology. Adequate specimens include eye swabs, swabs of vesicular lesions, saliva, throat swabs, cerebrospinal fluid, and tissues, as dictated by the clinical symptoms. Urine may also be a valid specimen.

      VZV causes two different clinical syndromes: chickenpox (varicella) - primarily in children, and shingles (zoster) - predominantly in adults (8-10). The most notable feature of varicella is a generalized vesicular rash which is usually accompanied by fever. Complications, such as pneumonia, can occur in neonates and in immunocompromised patients with a mortality of 10 to 40%.

      Zoster occurs as a reactivation of the latent varicella virus, and can result from almost any stimulus, such as fear, anxiety, etc. Pregnant women are at greater risk for varicella pneumonia than other adults. Postherpetic neuralgia is another complication of both varicella and zoster and is a significantly greater problem in immunosuppressed patients.

      As with HSV, acyclovir is the drug of choice for reducing symptoms due to VZV infection (8,11,12). In 1995, a varicella vaccine was approved for use in the United States; this vaccine gives a 70 to 85% protection rate in children (10,13,14).

      VZV can be isolated from clinical specimens, usually skin lesions, but also lung, eye, throat, and vesicular fluid in human diploid fibroblast cell cultures. Its cytopathology is fairly distinct from that of HSV. However, the virus is more labile than HSV and may not grow in culture unless care is taken in transporting the specimen to the lab.

      The purpose of combining HSV and VZV antibodies in a single test reagent is that both viruses can cause many of the same symptoms and clinical findings. HSV must be identified to prevent the spread of the virus to neonates at birth and to prevent spread of the venereal disease to other adults. Specific identification of the virus in conjunctival specimens allows prompt treatment with acyclovir to reduce the chance of blindness.
      Materials Required but Not Delivered· Acetone, reagent grade or better; stored in glass

      · Deionized or distilled water

      · Positive controls, for culture isolation procedures (reference HSV and VZV 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 other HSV/VZV permissive cell line

      · Tissue culture media (RPMI or Eagle's Minimum Essential Medium (EMEM) with precolostral bovine serum (FBS) and antibiotics or equivalent)

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

      · Sterile PBS, pH 7.0 to 7.6

      · Microscope slides, non-fluorescing

      · No. 1 coverslips

      · Aspirator device with disposable sterile Pasteur pipettes

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

      · Cytocentrifuge capable of depositing cell suspensions on slides

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

      · Optional: Filter combination for TRITC (excitation peak = 550nm, emission peak = 570nm) NOTE: Switching from a FITC to a TRITC filter will cause apple green fluorescence to disappear and yellow-green fluorescence to fluoresce hot pink, aiding in confirmation of non-specific staining.

      · 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
      • SimulFluor® HSV/VZV - (Catalog No. 5235). One 5 mL dropper vial containing monoclonal antibodies specific for HSV antigen and specific for VZV antigen, protein stabilizer, Evans blue and 0.1% sodium azide (preservative).
      • HSV Control Slides - (Catalog No. 5093). Two slides containing one well of HSV -1 infected HEp-2 cells, one well of HSV -2 infected cells, and one well of uninfected cells.
      • VZV Control Slides - (Catalog No. 5088). Two slides containing one well of VZV (clinical isolate) infected human foreskin fibroblasts and one well of uninfected human foreskin fibroblasts.
      • 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
      FormatSimulFluor
      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 ConditionsWhen stored at 2-8°C, the SimulFluor® HSV/VZV Immunofluorescence Assay 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

      · The sodium azide (NaN3) used as a preservative in the SimulFluor® reagent, PBS/Tween, and Mounting Fluid is toxic if ingested. Sodium azide may react with lead and copper plumbing to form highly explosive metal azides (15,16). Upon disposal, flush with large volumes of water to prevent build-up in plumbing.

      · Pooling or alteration of any reagent may cause erroneous results.

      · Do not substitute reagents from other manufacturers.

      · Incubation times or temperatures other than those specified may give erroneous results. Any such change must be validated by the user.

      · Do not allow shell vials or slides to dry at any time during the staining procedure.

      · Handle all specimens and materials, coming in contact with them as potentially infectious and dispose of with proper precautions. Decontaminate with 0.05% sodium hypochlorite.

      · Do not mouth pipette reagents.

      · 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 SimulFluor® reagent) as it is a potential carcinogen. If skin contact occurs, flush with large volumes of water.

      · 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 Size5 mL
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      SDS

      Title

      Safety Data Sheet (SDS) 

      References

      Reference overviewPub Med ID
      Biological and ultrastructural properties of acelagraft, a freeze-dried γ-irradiated human amniotic membrane.
      Lim LS, Poh RW, Riau AK, Beuerman RW, Tan D, Mehta JS
      Arch Ophthalmol 128 1303-10. 2010

      Show Abstract
      20938000 20938000
      Evaluation of light diagnostics Simulfluor HSV/VZV immunofluorescence assay
      Scicchitano LM, et.al.
      Diagn. Microbiol. Infect. Dis., 35:205-8 (1999) 1999

      10626130 10626130
      Role of bcl-2 and IL-5 in the regulation of anti-IgM-induced growth arrest and apoptosis in immature B cell lines. A cooperative regulation model for B cell clonal deletion.
      H Kamesaki, J A Zwiebel, J C Reed, J Cossman
      Journal of immunology (Baltimore, Md. : 1950) 152 3294-305 1994

      Show Abstract
      8144916 8144916

      Brochure

      Title
      Light Diagnostics™ products for infectious disease

      User Guides

      Title
      LIGHT DIAGNOSTICS™ KIT DFA SimulFluor HSV/VZV multi languages
      LIGHT DIAGNOSTICS™ SimulFluor HSV/VZV DFA KIT