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3293 | LIGHT DIAGNOSTICS™ SimulFluor® HSV 1/2, ~125 tests

3293
5 mL  
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      Key ApplicationsFormatHostDetection Methods
      IF SimulFluor M Fluorescent
      Description
      Catalogue Number3293
      Brand Family Chemicon®
      Trade Name
      • LIGHT DIAGNOSTICS
      • SimulFluor
      • Chemicon
      DescriptionLIGHT DIAGNOSTICS™ SimulFluor® HSV 1/2, ~125 tests
      OverviewLight Diagnostics SimulFluor® HSV 1/2 Immunofluorescence Assay is a direct immunofluorescence test intended for the detection and identification of herpes simplex virus type 1 (HSV-1) or herpes simplex virus type 2 (HSV-2) following amplification in cell culture or by direct examination of clinical specimens prepared by cytospin. Specimens found to be negative on direct specimen examination should be tested by cell culture.

      For in vitro diagnostic use.

      Test Principle:

      Light Diagnostics SimulFluor® HSV 1/2 Immunofluorescence Assay utilizes a single reagent for the simultaneous detection and identification of HSV-1 and HSV-2. The SimulFluor® HSV 1/2 Reagent consists of two components; the primary component specific to HSV-1 will bind to the glycoprotein C and a capsid-associated protein in HSV-1 infected cells, while the secondary component, specific for HSV-2, will bind to the glycoprotein G in HSV-2 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 an FITC filter set is used, HSV-1 infected cells will exhibit apple-green fluorescence and HSV-2 infected cells will exhibit yellow-gold fluorescence. The uninfected cells will stain a dull red due to the presence of Evans blue in the SimulFluor® HSV 1/2 reagent.

      Summary and Explanation :

      The following narrative does not necessarily imply a different Intended Use or additional inclusion in performance testing with this assay.

      Herpes simplex virus (HSV) is a member of the Herpesviridae family, alphaherpesvirus subfamily. HSV is a large, enveloped virus, about 190 nm in diameter containing a linear, double-stranded DNA enclosed within a baggy, phospholipoprotein envelope. 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 DNAs.

      HSV causes a multitude of human diseases (1-5). HSV-1 causes gingivostomatitis, intense pharyngitis, tonsillitis, and occasionally encephalitis in infants and children during their primary infections, and ocular, nasal, orolabial, and oropharyngeal lesions in 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, and is a formidable STD agent. If the virus is present in the birth canal at the time of delivery, either from primary or recurrent infection, the result will likely be a severe generalized infection in the neonate. Thus, maternal genital HSV infections pose a substantial risk to the fetus and newborn.

      Recurrent infection is the most common form of infection during gestation. Shedding of virus at the time of delivery is the usual route of transmission from mother to neonate. Neonatal HSV infection is generally symptomatic and often lethal, 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.

      Most HSV infections are characterized by infection of mucocutaneous surfaces and transport to 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 site of infection, with or without the associated clinical signs and symptoms. Recurrent lesions are usually less severe than the primary infection. Since recurrent episodes are more common in HSV-2 infected patients than those infected with HSV-1, typing can be of prognostic value (13).

      In immunocompromised patients such as those with AIDS and persons recovering from cancer or organ transplant treatments, HSV can cause painful recurrent lesions or overwhelming disseminated disease. Both serotypes can constitute a serious nosocomial problem for immunocompromised patients.

      The initial finding of HSV-1 in ocular and respiratory infections and HSV-2 in genital infections is still relatively true, but sufficient cases of HSV-2 in nongenital lesions and HSV-1 in genital lesions occur as to make testing for both types necessary. It is important to identify the particular herpesvirus and to differentiate between them in order to institute the proper epidemiological control measures and treatment. In addition, specific identification will differentiate HSV from other viruses, mycoplasmas, and bacteria which can cause similar clinical findings but would require different treatment strategies.

      Acyclovir, famciclovir, foscarnet, and other nucleoside analogs can reduce clinical symptoms and virus shedding in oral and genital herpes, encephalitis, neonatal herpes, and herpetic keratitis (6-8). Identification of the virus in conjunctival specimens allows prompt treatment with acyclovir to reduce the chance of blindness. Vaccines prepared with viral glycoproteins have been studied but not proven effective (9).

      HSV can be readily recovered from clinical specimens by passage in many cells lines, such as HEp2, HEK, NCI-H292, RD, MRC-5, mink lung, and others. Incubation time in stationary or roller cultures is from 1 to 5 days, and the cytopathology is quite evident. Tests on direct specimens include IFA, EIA, histopathology (1,2). DNA hybridization and PCR have been used by researchers to identify virus (12). 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.
      Materials Required but Not Delivered· Acetone, reagent grade; stored in glass

      · Deionized or distilled water

      · Positive controls, for culture isolation procedures (reference HSV strains available from ATCC, Rockville, MD.)

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

      · Sterile 1 dram shell vials with 12 mm coverslips for growth of MRC-5 or other HSV-permissive cell line

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

      · Dacron or cotton swabs (not alginate) for specimen collection

      · Viral transport medium (VTM) which is non-inhibitory to HSV, Hanks Balanced Salt Solution (HBSS) with antibiotics or equivalent

      · Sterile PBS (pH 7.0 - 7.6)

      · 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, appropriate filter combination for FITC (excitation peak = 490 nm, emission peak = 520nm), 100x, 200x and 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
      • SimulFluor® HSV1/2 Reagent - (Catalog No. 5243). One 5 mL dropper vial containing two components specific for HSV-1 and HSV-2, 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 cells, one well of HSV-2 infected cells and one well of uninfected cells.
      • 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 Light Diagnostics SimulFluor® HSV 1/2 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® HSV 1/2 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 (14, 15). 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.

      · 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.

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

      · Do not mouth pipette reagents.

      · Avoid contact with Evans blue (present in 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