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3129 | LIGHT DIAGNOSTICS™ SimulFluor® RSV/Flu A, ~125 tests

3129
5 mL  
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      Key ApplicationsFormatHostDetection Methods
      IF SimulFluor M Fluorescent
      Description
      Catalogue Number3129
      Brand Family Chemicon®
      Trade Name
      • LIGHT DIAGNOSTICS
      • SimulFluor
      • Chemicon
      DescriptionLIGHT DIAGNOSTICS™ SimulFluor® RSV/Flu A, ~125 tests
      OverviewIntended Use
      The Light Diagnostics™ SimulFluor® RSV/Flu A Immunofluorescence Assay is intended for use in respiratory specimens such as throat, nasal and nasopharyngeal swabs, throat washes, aspirates, broncho-alveolar lavages from patients with febrile respiratory illness following amplification of virus in cell culture. Negative results do not preclude influenza virus infection and should not be used as the sole basis for treatment or other management decision.
      Performance characteristic for influenza A were established when influenza A/H3 and A/H1 were the predominant influenza A viruses in circulation. When other influenza A viruses are emerging, performance characteristics may vary.
      If infection with a novel influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection control precautions for novel virulent influenza viruses and sent to state or local health department for testing. Viral culture should not be attempted in these cases unless a Biosafety Level 3 (BSL 3+) facility is available to receive and culture specimens.
      For In Vitro Diagnostic Use.

      Summary and Explanation
      Respiratory syncytial virus (RSV) occupies the genus Pneumovirus in the family Paramyxoviridae. It is a pleomorphic virus ranging from approximately 100 to 300 nm in diameter containing a linear, non-segmented, single-stranded, negative sense RNA. The envelope contains projections on the outer surface composed of two highly antigentic glycoproteins, F and G, and a major structural protein, N, in the nucleocapsid. The F, G, and N structural proteins divide RSV into two major antigenic groups; the "prototype" strains are Long (group A) and CH-18537 (group B) (1-4). RSV causes major epidemics of severe respiratory illness during the winter and spring seasons throughout the world. Because natural immunity to RSV is weak, these epidemics tend to occur every or every other year.
      RSV can be isolated from nasopharyngeal aspirates, nose or throat swabs, or lung specimens taken within the first few days of illness. The virus grows in human epithelial cells such as HEp2, primary rhesus monkey kidney (RMK),

      A549, MRC-5, and NCI-H292. Rapid and direct tests on suitable clinical specimens are diagnostic of RSV infections provided that standardized reagent and adequate controls are used. Confirmation by virus isolation is desired wherever possible.
      Hospitalized RSV-infected infants have been successfully treated with ribavirin, administered as an aerosol. However, this requires specialized care, and is not given routinely (4). Immune globulin is also available for treatment. RSV vaccines have a long history of failure but are still under intense development.
      Influenza A virus is a member of the family of Orthomyxoviridae. It is a large, enveloped virus, 110 nm in diameter, and containing a segmented, single-stranded RNA, with hemagglutinin (HA) and neuraminidase (NA) projections protruding through the glycoprotein membrane (5-7). Influenza A has a high frequency of mutation and cause periodic epidemics of influenza worldwide. Epidemics are particularly severe when the mutations have resulted in dramatic shifts in the HA or NA structure such that circulating antibodies in a community of people who do not recognize the virus strains (6-8). Specificity in influenza viruses is conferred by antigenic differences in two of the major structural proteins, the internal nucleoprotein (NP) and the matrix protein (M) (5-8).
      Influenza infections are characterized by tracheobronchitis, pharyngitis, myalgia, fever, headache, and malaise (5,6,9). Minimal coryza often distinguishes influenza from other viral respiratory illnesses. It is highly contagious among adults, being spread by aerosolized droplets and fomites. As with RSV, an incubation period of 1 to 4 days helps the virus spread rapidly within communities and within hospital wards. The most significant complication of influenza is pneumonia, which occurs by itself or may be mixed with bacterial pneumonia. Pneumonia, occurs most frequently in the elderly, in patients with weakened immune systems, or in patients with underlying diabetes, heart or lung disease, or chronic kidney disease. Children are more likely to experience additional symptoms of gastrointestinal pain, vomiting, myositis, otitis media, conjunctivitis, and croup.
      The increase in deaths from pneumonia during "flu season" is assumed to be due to influenza virus and is used to track influenza epidemics and to check the efficacy of influenza vaccines. Less common complications in adults include Reye's syndrome or other CNS involvement, cardiac symptoms, sinusitis, and otitis media.
      Amantadine and its analog, rimantadine, are effective in preventing up to 90% of influenza A infections and 100% of illnesses if taken prophylactically (5,10). They are also effective therapeutically by reducing the duration of illness if taken

      within the first 2 days of illness. Ribavirin may be effective in treating influenza A when administered by an aerosol route. Vaccines against influenza A constitute a major public health effort as new strains are identified. Current influenza vaccines generally have an efficacy rate of 70-90% (5,7).
      Influenza viruses can be grown in RMK, MDCK (Madin-Darby canine kidney), and occasionally other cell lines, depending on the particular strain of virus
      (11-13). Trypsin added to the culture fluid at ~2 µg/mL concentration greatly aids in influenza virus isolation in MDCK cells.
      Both standard culture and shell vials may be used. Direct tests, including immunofluorescence, (FA) and enzyme immunoassays (EIA) (11,14-17).
      Adequate specimens include nasal washes, throat swabs, and broncholaveolar lavages. Any direct test with standardized reagents is diagnostic of influenza infection, especially if confirmed by virus isolation.

      Test Principle
      Light Diagnostics™ SimulFluor® RSV/Flu A Immunofluorescence Assay utilizes a single reagent for the simultaneous detection and identification of RSV and influenza A. The primary component, specific for RSV, will bind to RSV F and G antigens in RSV-infected cells. The secondary component, specific for influenza A, will bind to influenza A nucleoprotein in influenza A-infected cells. Unbound reagent is removed by rinsing with phosphate buffered saline (PBS). The antigen-antibody complex can be visualized by fluorescence microscopy. The RSV antigen-antibody complex will exhibit an apple-green fluorescence and the influenza A antigen-antibody complex will appear yellow-gold. Uninfected cells stain a dull red due to the presence of Evans blue in the reagent.
      Materials Required but Not Delivered• Acetone, reagent grade or better; stored in glass
      • Deionized or distilled water
      • Virus culture controls (reference RSV and influenza A strains available from ATCC, Rockville, MD.)
      • Sodium hypochlorite solution, 0.05% (1:100 dilution of household bleach)
      • RSV and influenza susceptible cell lines such as HEp-2, MRC-5, primary monkey kidney (PMK), MDCK cell lines (14,15,16), or other influenza susceptible cell lines (e.g. LLC-MK2, etc.) Appropriate cell lines can be obtained from the American Type Culture Collection (ATCC), 12301 Parklawn Drive, Rockville, MD 20852.
      • Tissue culture media such as RPMI or Eagle's Minimum Essential Medium (EMEM) with fetal bovine serum (FBS) and antibiotics, or equivalent
      • Viral transport medium, which is non-inhibitory to RSV or influenza A (Hanks balanced salt solution (HBSS) with antibiotics, or equivalent)
      • Acetone cleaned glass 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
      • Fluorescence microscope with 100 watt mercury or halogen lamp, appropriate filter combination for FITC (excitation peak = 490 nm, emission peak = 520 nm), 160 to 200x and 400x magnification (dry objective)
      • Optional: filter combination for TRITC (excitation peak = 550 nm, emission peak = 570 nm)
      • 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® RSV/Flu A Reagent - (Catalog No. 5245).
      • RSV Control Slides - (Catalog No. 5012).
      • Influenza A/B Control Slides - (Catalog No. 5010).
      • Phosphate-Buffered Saline (PBS) - (Catalog No. 5087).
      • Tween 20/Sodium Azide Solution (100X) - (Catalog No. 5037).
      • Mounting Fluid - (Catalog No. 5013).
      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° to 8°C, the SimulFluor® RSV/Flu A Immunofluorescence Assay 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
      * The sodium azide 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. 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.

      * 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 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.
      • If infection with a novel influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection control precautions for novel virulent influenza viruses and sent to state or local health department for testing. Viral culture should not be attempted in these cases unless a Biosafety Level 3 (BSL 3+) facility is available to receive and culture specimens.
      Packaging Information
      Material Size5 mL
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      MSDS

      Title

      Safety Data Sheet (SDS) 

      References

      Reference overviewPub Med ID
      Type- and subtype-specific detection of influenza viruses in clinical specimens by rapid culture assay.
      Ziegler, T, et al.
      J. Clin. Microbiol., 33: 318-21 (1995) 1995

      Kivonat megmutatása
      7714186 7714186
      Prospective application of reverse transcriptase polymerase chain reaction for diagnosing influenza infections in respiratory samples from a children's hospital.
      Claas, E C, et al.
      J. Clin. Microbiol., 31: 2218-21 (1993) 1993

      Kivonat megmutatása
      8370755 8370755
      Genetic and antigenic analyses of influenza A (H1N1) viruses, 1986-1991
      Xu, X, et al
      Virus Res, 28:37-55 (1993) 1993

      8493812 8493812
      Comparison of rapid immunofluorescence assay to cell culture isolation for the detection of influenza A and B viruses in nasopharyngeal secretions from infants and children
      Spada, B, et al
      J Virol Methods, 33:305-10 (1991) 1991

      1783676 1783676
      Prevention, management, and control of influenza. Role of amantadine
      Mostow, S R
      Am J Med, 82:35-41 (1987) 1987

      3296750 3296750
      Rapid detection of influenza virus by shell vial assay with monoclonal antibodies
      Espy, M J, et al
      J Clin Microbiol, 24:677-9 (1986) 1986

      3533980 3533980

      Brochure

      Title
      Light Diagnostics™ products for infectious disease

      User Guides

      Title
      LIGHT DIAGNOSTICS™ SimulFluor® RSV/FLU A DFA KIT
      LIGHT DIAGNOSTICS™ SimulFluorÒ RSV/FLU A DFA KIT