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| Catalog # | Availability | Size | Quantity | Unit Price | Save For Later Wish List | |
|---|---|---|---|---|---|---|
| EKCA1028-1 | 2 weeks | 96 T | call |
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| Name | Mumps IgM ELISA Kit | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| INTENDED USE | The Calbiotech, Inc. (CBI) Mumps IgM ELISA tests system is an enzyme linked immunosorbent assay (ELISA) for the detection of IgM class antibodies to Mumps in human serum or plasma. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| SUMMARY AND EXPLANATION | Infection with Mumps virus causes fever, headache, and swelling and tenderness of the salivary glands. Most adults born before 1957 have been infected naturally and are probably immune. Mumps can occur in unimmunized children, or adolescents and young adults who graduated from school prior to the law requiring mumps immunization. About 1/3 of people have no symptoms. The first symptoms usually appear 16 to 18 days after exposure. It begins with fever and pain upon opening the mouth or eating. Possible complications include meningitis (swelling of the covering of the brain and spinal cord), encephalitis (swelling of the brain), deafness, and in adult males, swelling of the testicles. The virus may cause a miscarriage if a woman becomes infected during the first three months of pregnancy. Mumps IgM antibodies by ELISA are present in serum of 72% of patients by day 2 of clinical illness and in essentially all patients after day 5. A significant increase in titer of mumps IgG by ELISA is found in over 90% of paired acute and convalescent mumps sera in which mumps IgM antibodies can also be found. Increases in mumps antibody titers in paired acute and convalescent sera are valuable for confirmation of acute infection even in the presence of specific IgM antibodies because 50% of patients still have elevated levels of reactive IgM 5 or more months after clinical mumps. In mumps meningitis, the Mumps IgG Antibody Index is increased in about 83% of patients and the Mumps IgM Antibody Index is increased in about 67% of those with detectable IgM in the CSF. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| PRINCIPLE OF THE TEST | Diluted patient serum (serum diluent contains sorbent to remove Rheumatoid Factor and human IgG interference) is added to wells coated with purified antigen. IgM specific antibody, if present, binds to the antigen. All unbound materials are washed away and the enzyme conjugate is added to bind to the antibody-antigen complex, if present. Excess enzyme conjugate is washed off and substrate is added. The plate is incubated to allow the hydrolysis of the substrate by the enzyme. The intensity of the color generated is proportional to the amount of IgM specific antibody in the sample.
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| MATERIALS NOT PROVIDED | Distilled or deionized water Precision pipettes Disposable pipette tips ELISA reader capable of reading absorbance at 450nm Absorbance paper or paper towel Graph Paper | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| STORAGE AND STABILITY | Store the kit at 2-8 C. Keep microwells sealed in a dry bag with desiccants. The reagents are stable until expiration of the kit. Do not expose test reagents to heat, sun or strong light | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| WARNINGS AND PRECAUTIONS | Potential biohazardous materials: The calibrator and controls contain human source components which have been tested and found non-reactive for hepatitis B surface antigen as well as HIV antibody with FDA licensed reagents. However, there is no test method that can offer complete assurance that HIV, Hepatitis B virus or other infectious agents are absent. These reagents should be handled at the Biosafety Level 2, as recommended in the Centers for Disease Control/National Institutes of Health manual, "Biosafety in Microbiological and Biomedical Laboratories." 1984. Optimal results will be obtained by strict adherence to the test protocol. Precise pipetting as well as following the exact time and temperature requirements is essential. Do not pipette by mouth. Do not smoke, eat, or drink in the areas in which specimens or kit reagents are handled. The components in this kit are intended for use as an integral unit. The components of different lots should not be mixed. Control sera and sample diluent contain preserved with sodium azide. Sodium azide may react with lead and copper plumbing to form explosive metal azide. On disposal, flush with a large volume of water. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| SPECIMEN COLLECTION AND HANDLING | 1. Collect blood specimens and separate the serum. 2. Specimens may be refrigerated at 2–8 C for up to seven days or frozen for up to six months. Avoid repetitive freezing and thawing. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| REAGENT PREPARATION | Prepare 1X Wash buffer by adding the contents of the bottle (25 ml, 20X) to 475 ml of distilled or deionized water. Store at room temperature (20-25C). | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| ASSAY PROCEDURE | Bring all specimens and kit reagents to room temperature (20-25C) and gently mix. Place the desired number of coated strips into the holder. Negative control, positive control, and calibrator are ready to use. Prepare 1:21 dilution of test samples, by adding 10 l of the sample to 200 l of sample diluent. Mix well. Dispense 100 l of diluted sera, calibrator and controls into the appropriate wells. For the reagent blank, dispense 100l sample diluent in 1A well position. Tap the holder to remove air bubbles from the liquid and mix well. Incubate for 20 minutes at room temperature. Remove liquid from all wells. Wash wells three times with 300 l of 1X wash buffer. Blot on absorbance paper or paper towel. Dispense 100 l of enzyme conjugate to each well and incubate for 20 minutes at room temperature. Remove enzyme conjugate from all wells. Wash wells three times with 300 l of 1X wash buffer. Blot on absorbance paper or paper towel. Dispense 100 l of TMB substrate and incubate for 10 minutes at room temperature. Add 100 l of stop solution. Read O.D. at 450 nm using ELISA reader within 15 min. A dual wavelength is recommended with reference filter of 600-650 nm. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| CALCULATION OF RESULTS | Check Calibrator Factor (CF) value on the calibrator bottle. This value might vary from lot to lot. Make sure you check the value on every kit. Calculate the cut-off value: Calibrator OD x Calibrator Factor (CF). Calculate the Ab (Antibody) Index of each determination by dividing the O.D. value of each sample by cut-off value. Example of typical results: Calibrator mean OD = 0.8 Calibrator Factor (CF) = 0.5 Cut-off Value = 0.8 x 0.5= 0.400 Positive control O.D. = 1.2 Ab Index = 1.2 / 0.4 = 3 Patient sample O.D. = 1.6 Ab Index = 1.6 / 0.4 = 4.0 | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| LIMITATIONS OF THE TEST | To enhance sensitivity and specificity of this IgM test provided sample diluent has been formulated to block IgG and Rheumatoid Factor (RF) interferences. Turbidity could be seen after diluting serum with sample diluent. This turbidity is due to the blocking of serum IgG and shows no interference with test results. It can be removed by centrifugation. In specimens with high RF and high autoimmune antibodies, the possibility of eliminating the interferences cannot be ruled out entirely. The test results obtained using this kit serve only as an aid to diagnosis and should be interpreted in relation to the patient’s history, physical findings and other diagnostic procedures. Lipemic or hemolyzed samples may cause erroneous results. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| QUALITY CONTROL | The test run may be considered valid provided the following criteria are met: The O.D. of the Calibrator should be greater than 0.250. The Ab index for Negative control should be less than 0.9. 3. The Ab Index for Positive control should fall within the range specified on the COA/label. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| INTERPRETATION | The following is intended as a guide to interpretation of Mumps IgM test results; each laboratory is encouraged to establish its own criteria for test interpretation based on sample populations encountered. Antibody Index Interpretation <0.9 No detectable antibody to Mumps IgM by ELISA. Borderline positive. Follow-up testing is recommended if clinically indicated. >1.1 Detectable antibody to Mumps IgM by ELISA. | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| PERFORMANCE CHARACTERISTICS | 1. Sensitivity and Specificity 129 patient sera were tested by this Mumps IgM ELISA and a reference ELISA method. 19 sera were positive and 105 were negative by both methods (96% agreement). The results are summarized below:
2. Precision Intra-Assay Study
Inter-Assay Study
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| REFERENCES | Davidkin I; Valle M; Julkunen I. Persistence of anti-mumps virus antibodies after a two-dose MMR vaccination. A nine-year follow-up.Vaccine 1995;13(16):1617-22. Chomel JJ; Robin Y; Durdilly R; Thouvenot D; Langlois M; Aymard M.Rapid direct diagnosis of mumps meningitis by ELISA capture technique. J Virol Methods 1997;68(1):97-104. Nigro G; Nanni F; Midulla M. Determination of vaccine-induced and naturally acquired class-specific mumps antibodies by two indirect enzyme-linked immunosorbent assays. J Virol Methods 1986;13(2):91-106. Harmsen T; Jongerius MC; van der Zwan CW; Plantinga AD; Kraaijeveld CA; Berbers GA. Comparison of a neutralization enzyme immunoassay and an enzyme-linked immunosorbent assay for evaluation of immune status of children vaccinated for mumps. J Clin Microbiol 1992;30(8):2139-44. Novotn´y J. Properties and use of mumps viral antigen for detection of specific IgG and IgM antibodies in enzyme-linked immunosorbent assay. Acta Virol 1990;34(6):574-7. Johnson CE; Kumar ML; Whitwell JK; Staehle BO; Rome LP; Dinakar C; Hurni W; Nalin DR. Antibody persistence after primary measles-mumps-rubella vaccine and response to a second dose given at four to six vs. eleven to thirteen years. Pediatr Infect Dis J 1996; 15(8):687-92. |
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