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|Sensitivity:||99.84%||Application:||Legionella Pneumophila IgG/ IgM|
Legionella pneumophila IgG/ IgM tests ELISA Test, Elisa Sandwich method, for quantitative measurement
Product Name: Legionella pneumophila IgG/ IgM tests ELISA Test Kit
The Legionella pneumophila 1-7 IgG and IgM tests are qualitative immunoassays for the demonstration of human antibodies directed against Legionella pneumophila serotypes 1 to 7. The assays are recommended to complement the findings of direct detection methods and for differential diagnosis in case of atypical pneumonia.
Legionella belongs to the species of legionellaceae, genus Legionella. It is a gram negative, aerobic, pleomorphic, non-spore forming bacillus which is motile by means of a single or multiple polar or subpolar flagella. Currently, the genus includes 39 Legionella species. Immunologic diversity within species is reflected in the creation of serogroups. The most important human pathogen is the species Legionella pneumophila which comprises 14 serologic groups.
Legionella are ubiquitous bacteria which reside in surface and drinking water and multiply in amoebae and other protozoa. Warm water systems with a temperature of 25-45°C are ideal for multiplication. After transmission to humans in aerosols, the bacteria are phagocytized by alveolar macrophages. So called evasion mechanisms enable bacteria not only to survive but also to multiply within a phagocytic cell.
Due to non-specific clinical symptoms, diagnosis is based on laboratory techniques such as direct pathogen detection, derived antigen detection, or detection of specific antibodies.
Direct pathogen detection in culture or in direct immunofluorescence assays (DFA) are of outstanding importance because fast diagnosis is often decisive for the patient. Sputum, tracheobronchial secretion, and pleura punctates are ideal samples. However, sensitivities of only 50-60 % are described for both techniques.
To compensate for the above mentioned drawbacks of direct detection methods, serologic test systems are widely used. Since multiple serogroups of L. pneumophila are human pathogens, pool antigens are favored. In the SERION ELISA classic IgG/IgM, a mixture of serogroups 1-7 is bound to the solid phase
|Plate: 96 wells configured in twelve 1X8-well||strips coated with a|
|formalin-inactivated sonicated preparation of L. pneumophila Groups 1-6 antigens.||1 plate|
|The strips are packaged in a strip holder and sealed in an envelope with desiccant.|
|Conjugate: Conjugated(horseradish peroxidase) goat||anti-human||IgG/IgA/IgM.||15mL|
|Ready to use. A white cap.|
|Positive Control (Monkey Serum): A red cap.||0.35mL|
|Calibrator (Monkey Serum): A blue cap.||0.5mL|
|Negative Control (Human Serum): A green cap||0.35mL|
|Abnova (Sample Diluent): A green cap containing Tween-20, bovine serum|
|albumin and phosphate-buffered-saline, (pH7.2±0.2). Ready to use. Note: Shake||30mL|
|Well Before Use.|
|TMB: An amber cap containing 3,3’,5,5’-tetramethylbenzidine(TMB). Ready to use.||15mL|
|Stop solution: Red cap containing 1M H2SO4, 0.7M HCl. Ready to use||15mL|
|Wash buffer concentrate (10X): dilute 1 part concentrate + 9 parts deionized or|
|phosphate-buffered-saline and solution (Blue solution). Note: 1*solution will have a|
|pH of 7.2±0.2.|
Each kit contains the following components in sufficient quantities to perform the number of tests indicated on packaging label. Note: All reactive reagents contain sodium azide as a preservative at a concentration of 0.1% (w/v).
The following components are not kit lot number dependent and may be used interchangeably with the ELISA assays: TMB, Stop Solution, and Wash Buffer.
Note: Kit also contains:
Component list containing lot specific information is inside the kit box. Package insert providing instructions for use.
The ELISA (Enzyme Linked Immunosorbent Assay) is an immunoassay, which is particularly suited to the determination of antibodies in the field of infectious serology. The reaction is based on the specific interaction of antibodies with their corresponding antigen. The test strips of the microtiter plate are coated with specific antigens of the pathogen of interest. If antibodies in the sample are present, they bind to the fixed antigen. A secondary antibody, which has been conjugated with the enzyme alkaline phosphatase, detects and binds to the immune complex. The colourless substrate pnitrophenylphosphate is then converted into the coloured product p-nitrophenol. The signal intensity of this reaction product is proportional to the concentration of the analyte in the sample and is measured photometrically.
1. Remove the individual components from storage and allow them to warm to room temperature (20-25°C).
2. Determine the number of microwells needed. Allow six Control/Calibrator determinations (one Blank, one Negative Control, three Calibrators and one Positive Control) per run. A Reagent Blank should be run on each assay. Check software and reader requirements for the correct Controls/Calibrator configurations. Return unused strips to the resealable pouch with desiccant, seal, and return to storage between 2 and 8
3. Prepare a 1:21 dilution (e.g.: 10µL of serum + 200µL of Sample Diluent. NOTE: Shake Well Before Use) of the Negative Control, Calibrator, Positive Control, and each patient serum. The Sample Diluent will undergo a color change confirming that the specimen has been combined with the diluent.
4. To individual wells, add 100µL of each diluted control, calibrator and sample. Ensure that the samples are properly mixed. Use a different pipette tip for each sample.
5. Add 100µL of Sample Diluent to well A1 as a reagent blank. Check software and reader requirements for the correct reagent blank well configuration.
6. Incubate the plate at room temperature (20-25°C) for 25± 5 minutes.
7. Wash the microwell strips 5X.
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|Email:||Jerry @ newlifebiotest .com|
Contact Person: Jerry Meng