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3121 | LIGHT DIAGNOSTICS™ SimulFluor® Flu A/Flu B, Flu A & Flu B, ~50 tests

2 mL  
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      Key Spec Table

      Key ApplicationsFormatHostDetection Methods
      Catalogue Number3121
      Brand Family Chemicon®
      Trade Name
      • SimulFluor
      • Chemicon
      DescriptionLIGHT DIAGNOSTICS™ SimulFluor® Flu A/Flu B, Flu A & Flu B, ~50 tests
      OverviewThe Light Diagnostics SimulFluor® Flu A/Flu B Immunofluorescence Assay is intended for the detection and identification of influenza A and influenza B in respiratory specimens such as throat, nasal and nasopharyngeal swabs, nasopharyngeal aspirates, broncho-alveolar lavages from patients with febrile respiratory illness and following amplification of virus in cell culture. Specimens found to be negative on direct specimen examination must be confirmed with culture.

      For In Vitro Diagnostic Use.

      Test Principle

      Light Diagnostics SimulFluor® Flu A/Flu B Immunofluorescence Assay utilizes a single reagent for the simultaneous detection and identification of influenza A and influenza B. The primary component, specific for influenza A, will bind to influenza A nucleoprotein in influenza A-infected cells. The secondary component, specific for influenza B, will bind to influenza B nucleoprotein in influenza B-infected cells. Unbound reagent is removed by rinsing with phosphate-buffered saline (PBS). The complexes are visualized with a fluorescence microscope. The influenza A antigen-antibody complex will exhibit an apple-green fluorescence and the influenza B antigen-antibody complex will be yellow-gold. Uninfected cells stain a dull red due to the presence of Evans blue in the reagent.

      Summary and Explanation:

      Influenza A and B viruses are members of the Orthomyxoviridae family. They are large enveloped viruses, about 110 nm in diameter, with hemagglutinin (HA) and neuraminidase (NA) projections protruding through the glycoprotein membrane, and containing a segmented, single-stranded RNA (1,2). Both viruses have 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 do not recognize the virus strains (2,3). Influenza C virus causes only mild upper respiratory illness in children - never epidemics - and does not exhibit large antigenic shifts, partly because it does not contain neuraminidase. Specificity in all three influenza types is conferred by antigenic differences in two of the major structural proteins - the internal nucleoprotein (NP) and the matrix protein (M) (1-3).

      Influenza is characterized by tracheobronchitis, pharyngitis, myalgia, fever, headache, and malaise (1,2,4). Minimum coryza often distinguishes influenza from other viral respiratory illnesses. It is highly contagious, and spread by aerosolized droplets and fomites. An incubation period of 1-4 days helps the rapid spread of the virus within communities. The most significant compli-cation of influenza is pneumonia, which occurs most frequently in the elderly, in patients with weakened immune systems, or with chronic kidney disease. 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.

      Children often experience additional symptoms of gastrointestinal pain, vomiting, myositis, otitis media, conjunctivitis, and croup, and are more likely to show symptoms with influenza B virus rather than influenza A virus. In contrast, the more severe illnesses in adults, resulting in hospitalization, are likely to be caused by influenza A.

      Amantadine and its analog, rimantadine, are effective in preventing up to 90% of influenza A infections (not influenza B) and 100% of illnesses if taken prophylactically (1,5). They are also effective therapeutically by reducing the duration of illness if taken within the first 2 days of illness. Ribavirin, a nucleoside analog of guanosine, may be effective in treating influenza A and B when administered by an aerosol route.

      Influenza A and B can cause similar symptoms and have similar clinical presentations. All specimens in suspected influenza cases should be tested for both viruses . It is important to identify the particular virus for epidemiological considerations, because treatment is available for influenza A infections, and to avoid nosocomial spread of the virus. Further, identification of influenza viruses is necessary to differentiate them from other viruses. Mycoplasmas and bacteria can cause similar clinical findings but require different treatment strategies.

      Identification and differentiation of influenza A or B in cell culture or patient specimens is usually made using reagents containing antibodies specific for influenza A or influenza B (1). Influenza viruses can be isolated in primary rhesus monkey kidney cells (pRMK), Madin Darby canine kidney (MDCK) cells, and other cell lines, depending on the particular strain of virus (6-9). The addition of trypsin to the culture fluid at ~2 mg/mL concentration greatly aids in influenza virus isolation in MDCK cells. Since many influenza strains do not cause definable cytopathology, cultures must be tested by hemagglutination (HA) or hemadsorption (HAd ) with guinea pig or chicken erythrocytes to assess viral growth.
      Materials Required but Not Delivered· Acetone, reagent grade; stored in glass

      · Deionized or distilled water

      · Virus culture controls (reference influenza A and influenza B strains available from ATCC, Rockville, MD.)

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

      · Sterile standard tubes or shell vials with 12 mm coverslips for growth of primary monkey kidney (PMK), MDCKcell lines (14,15,16) or other influenza susceptible cell lines (e.g. MRC-5, LLC-MK2, etc.)

      · 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 influenza A or influenza B (Hanks balanced salt solution with antibiotics, or equivalent)

      · Acetone-cleaned glass 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 = 520 nm), 160-200x 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
      Product Information
      • SimulFluor® Flu A/Flu B - (Catalog No. 5250). One 2 mL dropper vial containing a primary component specific for influenza A nucleoprotein antigen and a secondary component specific for influenza B nucleoprotein antigen, protein stabilizer, Evans blue and 0.1% sodium azide (preservative).
      • Influenza A/B Control Slides - (Catalog No. 5010). Two slides containing one well of influenza A-infected cells, one well of influenza B-infected cells and two wells 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
      Key Applications
      • Immunofluorescence
      Biological Information
      Antibody TypeMonoclonal Antibody
      Physicochemical Information
      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® Flu A/Flu B 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 (NaN3) used as a preservative in the SimulFluor® reagent, PBS/Tween and Mounting Fluid is toxic if ingested. NaN3 may react with lead and copper plumbing to form highly explosive metal azides (17,18). 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.
      Packaging Information
      Material Size2 mL
      Transport Information
      Supplemental Information




      Safety Data Sheet (SDS) 


      Reference overviewPub Med ID
      Evaluation of R-Mix FreshCells in shell vials for detection of respiratory viruses.
      Fong, C K, et al.
      J. Clin. Microbiol., 38: 4660-2 (2000) 2000

      Show Abstract
      11101618 11101618


      Light Diagnostics™ products for infectious disease

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      SimulFluorÒ Flu A/Flu B