Components

Measles Reagent - (Catalog No. 5030). One (1) 2 mL vial containing measles monoclonal antibody.Measles Control Slides - (Catalog No. 5031). Two (2) slides each containing one well of measles virus infected cells (positive) and one well of noninfected cells (negative).Anti-Mouse IgG / FITC Conjugate - (Catalog No. 5008). One (1) 10 mL vial containing FITC labeled mouse IgG antibody and 0.02% Evans blue counterstain.PBS - (Catalog No. 5087). One (1) packet phosphate buffered saline salts yield 1 liter when dissolved in distilled water. Store in a clean closed container at room temperature.Tween 20 / Sodium Azide Solution (100X) - (Catalog No. 5037). One (1) 10 mL vial containing polyoxyethylene sorbitan monolaurate (Tween 20) and sodium azide (NaN3) concentrate to be diluted 1:100 in PBS.Mounting Fluid - (Catalog No. 5013). One (1) 10 mL dropper vial containing tris buffer, glycerin, fluorescence enhancer, and sodium azide as preservative. Store at room temperature.

Disclaimer

CE MarkFor in vitro Diagnostic Use

General description

Diagnostics Kit

Light Diagnostics Measles Kit is intended for the culture confirmation of measles virus infected cells.

For In Vitro Diagnostic Use

Test Principle:

The Light Diagnostics Measles Kit utilizes the indirect immunofluorescence assay for the detection of measles antigens. The assay utilizes a blend of monoclonal antibodies directed against matrix and hemagglutinin proteins of the measles virus. The antibodies will bind specifically to measles antigens present in the cells fixed in the substrate antigen wells. The antibodies are then detected with a secondary fluorescein isothiocyanate (FITC) labeled goat mouse antibody.

An apple green fluorescence is observed in the nucleus and cytoplasm of measles virus infected cells.

Summary And Explanation:

The measles virus causes an acute generalized infection and, until recently, has been one of the most common viral diseases of children8. Measles infection results in a febrile illness characterized by a prodrome similar to other respiratory tract infections. The characteristic clinical sign of measles, Koplik′s spots, appear during or after the prodrome on the mouth epithelium11. The most consistent and remarkable feature of in vivo and in vitro infection is the formation of multi-nucleated giant cells1. Complications of measles infection include otitis media, pneumonia, and rarely, subacute sclerosing panencephalitis (SSPE) which may develop after a latent period of 1 to 10 years11.

Measles is preventable through vaccination but recent epidemics have occurred in vaccinated populations even in developed countries9.

The measles virus belongs to the family Paramyxoviridae and is a member of the genus Morbillivirus10. It is an enveloped pleomorphic virus ranging from 100 to 250 nm in diameter with a single-stranded linear RNA.

The measles virus can be easily isolated in primary human or monkey kidney cells4. Measles giant cells can be demonstrated in respiratory secretions, urine, or epithelial surfaces such as pharynx, nasal and buccal mucosa, and conjunctiva2,3,5. Four fold increases in serum titers and ELISA techniques can be used for measles diagnosis. Higher sensitivity and specificity is obtained with immunofluorescence by demonstrating viral antigens6,7.

Legal Information

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

Physical form

Format: Purified

Storage and Stability

When stored at 2-8° C, the Measles 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:

1. The sodium azide (NaN3) used as a preservative in the reagent, Tween 20, 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.

2. Pooling or diluting reagents may cause erroneous results.

3. Do not use reagents after expiration.

4. Do not allow slides to dry at any time during the staining procedure.

5. Handle all specimens and materials coming in contact with them with the same precautions as for potentially infectious material. Decontaminate with 0.05% sodium hypochlorite (a 1:100 dilution of household bleach).

6. Avoid contact with Evans blue (present in the anti mouse IgG / FITC conjugate) as it is a potential carcinogen. If skin contact occurs, flush with large volumes of water.

7. Do not mouth pipette reagents.

8. Do not substitute reagents from other manufacturers.

9. Alteration of protocol provided may cause erroneous results.

10. When staining multiple samples on a slide avoid cross contamination between samples.

11. Control slides should be tested with each cell culture isolate. The positive control should show cells exhibiting fluorescence of the nucleus and cytoplasm. The negative control should show cells staining a dull red due to the Evans blue counterstain.

Caution: Fluorescent staining of cell fragments, often due to trapping of the conjugate in such debris, should be ignored. If the positive and negative controls cannot be clearly distinguished the test is invalid and should be rerun after checking the various sources of error mentioned in "Limitations". A negative result may be due to a variety of factors such as: inadequate sample, improper sample collection and handling, improper culture technique, use of inappropriate cell line or temperature during isolation, or factors mentioned in "Limitations". All negative results should be reported as "No virus observed.”