Application

Research CategoryInfectious Diseases

Research Sub CategoryInfectious Diseases - Viral

Components

hMPV Reagent - (Catalog No. 5091). One 2 mL dropper vial containing FITC-labeled anti-hMPV monoclonal antibodies in phosphate buffered saline (PBS), Tween 20, and 0.02% Evans blue, with sodium azide as a preservative.hMPV Control Slide - (Catalog No. 5092). Two control slides containing one hMPV type A and one hMPV type B infected (positive) well and one non-infected (negative) well.Phosphate Buffered Saline - (Catalog No. 5087) One packet of PBS salts to yield 1 liter upon reconstitution with distilled water. Store in a clean, closed container at room temperature.Tween 20 / Sodium Azide Solution (100 X) - (Catalog No. 5037). One 10 mL Tween 20 and sodium azide concentrate to be diluted 1:100 in PBS.Mounting Fluid - (Catalog No. 5013). One 10 mL dropper vial containing Tris buffer, glycerin, fluorescence enhancer, and sodium azide as preservative. Store at room temperature.

Disclaimer

For in vitro Diagnostic UseCE Mark

General description

Diagnostics Kit

Light Diagnostics™ Human Metapneumovirus Direct Immunofluorecsence Assay is intended for the detection and identification of human metapneumovirus (hMPV) from specimens collected using nasal swabs from patients exhibiting febrile respiratory illness.

For In Vitro Diagnostic Use.

Test Principle:

Light Diagnostics Human Metapneumovirus Direct Immunofluorescence Assay utilizes a direct immunofluorescent antibody technique for identifying hMPV in infected cell preparations made from respiratory specimens. The monoclonal antibodies are labeled with FITC (fluorescein isothiocyanate), which fluoresce apple-green when illuminated with ultraviolet light. The labeled antibody will bind to viral antigen present in the specimen. Unbound reagent is removed by washing with buffer. Cells in positive specimens will fluoresce apple-green while uninfected cells will stain dull red due to the presence of Evans blue.

Summary and Explanation:

Human metapneumovirus (hMPV) is a member of the pneumoviriniae subfamily of paramyxoviruses, first described in 2001 from pediatric respiratory specimens in the Netherlands. It is an enveloped pleomorphic virus with a single negative-strand RNA genome. Two major serotypes A and B have been described.

Several studies have identifed human metapneumovrius in specimens worldwide, and estimated that by 5 years of age, 70% of children have developed antibodies to human metapneumovirus. Although early studies focused on children, human metapneumovirus also affects the elderly and immunocompromised adults. Human metapneumovirus can cause upper and lower respiratory tract illness and is clinically similar to Respiratory syncitial virus (RSV). Commonly observed clinical symptoms include cough, fever, wheezing, and bronchiolitis. In addition human metapneumovirus may exacerbate asthma. Human metapneumovirus circulates seasonally and co-circulates with RSV.

Legal Information

CHEMICON is a registered trademark of Merck KGaA, Darmstadt, Germany

Storage and Stability

When stored at 2-8°C, the Human Metapneumovirus Direct Fluorecsence 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.

During incubation, slides should be protected from light and kept in a humid chamber at the recommended temperature. A marked decrease in fluorescence may indicate conjugate deterioration. A positive control should be tested with each specimen to ensure proper functioning of these reagents and proper staining procedure. If after appropriate analysis there is a decrease in staining intensity, discontinue use of the reagents.

Warnings and Precautions:

⋅ Sodium azide (present in the conjugate, diluent, PBS solution, and mounting fluid) is toxic if ingested. Sodium azide can react with lead or copper plumbing to form potentially explosive metal azides. When disposing of these materials, flush with large volumes of water to prevent azide build-up.

⋅ Pooling or diluting conjugates may cause erroneous results.

⋅ Avoid leaving reagents above 2-8°C for extended periods.

⋅ 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 allow slides to dry at any time during the staining procedure.

⋅ Handle all specimens and materials as potentially infectious material. Decontaminate with 0.05% sodium hypochlorite (a l:100 dilution of household bleach).

⋅ Do not expose reagents to bright light during storage or incubation.

⋅ Avoid contact with Evans blue as it is a potential carcinogen. If skin contact occurs, flush with large volumes of water.

⋅ Do not mouth pipette reagents.

⋅ Do not substitute reagents from other manufacturers.

⋅ Alteration of protocol provided may cause erroneous results.

⋅ When staining multiple samples on a slide, avoid cross-contamination of samples.