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
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
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
|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
|albumin and phosphate-buffered-saline, (pH7.2±0.2). Ready to use.
|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
|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
6. Incubate the plate at room temperature (20-25°C) for 25± 5
7. Wash the microwell strips 5X.
|ORIENT NEW LIFE MEDICAL CO., LTD.|
|Email:||Jerry @ newlifebiotest .com|