Mycoplasma pneumoniae IgG ELISA Kit

Sample

Serum, plasma

Species Reactivity

Human

Detection Method

iELISA

Intended Use

ELISA Mycoplasma pneumoniae IgG are quantitative and qualitative tests for detection of human antibodies in serum or plasma against Mycoplasma pneumoniae. For sale in the U.S. for Research Use Only. Not for use in diagnostic procedures.

Contents of Kit

1. Break apart microtiter test strips each with 8 antigen coated single wells (altogether 96), 1 frame, the coating material is inactivated. 12
2. Standard serum (ready-to-use), Human serum in phosphate buffer with protein; negative for anti-HIV-Ab, anti-HBs-Ag (Hepatitis B-Virus-surface antigen) and anti-HCV-Ab; preservative: < 0.1 % sodium azide, colouring: Amaranth O. 2 × 2 ml
3. Negative control serum (ready-to-use), Human serum in phosphate buffer with protein; negative for anti-HIV, anti-HBs (Hepatitis BVirus-surface antigen) and anti-HCV; preservative: < 0.1 % sodium azide, colouring: Lissamin green V. 2 ml
4. Anti-human-IgG-conjugate (ready-to-use), Anti-human-IgG from goat (polyclonal), conjugated to alkaline phosphatase, stabilized with protein stabilization solution preservative: 0.01 % methylisothiazolone, 0.01 % bromnitrodioxane.13 ml.
5. Washing solution concentrate (sufficient for 1 litre), Sodium chloride solution with Tween 20, 30 mM Tris, preservative: < 0.1 % sodium azide. 33.3 ml
6. Dilution buffer, Phosphate buffer with protein and Tween 20; preservative: < 0.1 % sodium azide 0.01 g/l Bromphenol blue sodium salt. 2 × 50 ml
7. Stopping solution, 1.2 N sodium hydroxide. 15 ml
8. Substrate (ready-to-use), Para-nitrophenylphosphate, solvent free buffer, preservative: < 0.1 % sodium azide, (Substrate in unopened bottle may have a slightly yellow color. This does not reduce the quality of the product!) 13 ml

Storage

Reagent	Storage	Stability
microtiter strips (antigen)	after opening at 2-8°C in closed aluminum bag with desiccant. Strips which are not used must be stored in the press-seal bag of aluminum compound foil under dry and airtight conditions!	4 weeks
control sera / standard sera	after opening at 2-8°C	until expiry date; 24 months from date of production
conjugate	ready-to-use solution, at 2-8°C, Avoid contamination (sterile tips!)	until expiry date 28 months from date of production
washing solution	concentrate after opening at 2-8°C; working dilution at 2-8°C; working dilution at room temperature. Bottles used for the working dilution should be cleaned regularly, discard cloudy solutions.	until expiry date; 2 weeks; 1 week
dilution buffer	after opening at 2-8°C, Discard cloudy solutions!; unopened	24 months; until expiry date; 36 months from date of production
substrate	ready-to-use solution at 2-8°C, protected from light! Avoid contamination (sterile tips!) Discard when solution turns yellow (extinction against distilled water > 0.25).	until expiry date 24 months from date of production
stopping solution	after opening at room temperature	until expiry date

General Description

Mycoplasma pneumoniae causes various respiratory diseases. Primary atypical pneumonia, pharyngitis and tracheobronchitis are the main diseases. In the USA, Mycoplasma pneumoniae is the cause of a significant number of cases of pneumonia every year.
Various symptoms complicate differentiation from respiratory infections of different genesis (e.g. RSV-, Influenza Virus-infections). Mycoplasma pneumoniae colonizes epithelia of the trachea, bronchia and bronchiolae. Early non-specific symptoms such as headache and fever occur, accompanied by a non-productive, dry cough. During the course of infection an interstitial pneumonia can develop. Since immunity is insufficient after infection, re-infections can occur. Superinfections with Mycoplasma pneumoniae after viral (e.g. measles) or bacterial infections (e.g. Streptococcus pneumoniae) have been observed.
Pathology of infections is based on adherence of the mycoplasmas to epithelia cells of the respiratory tract, on sliding motility over host cells, and pathologic hyperactivity of cellular immune responses of the host. Homologies between bacterial adhesins and adhesins of the host may prevent the host from developing an adherence inhibiting immune response that might prevent recolonization.
Due to the varying symptoms, diagnosis cannot usually be based on symptoms only, so pathogen detection or serological methods are employed. The Complement Fixation Test (CFT) is a common serological technique used for the detection of Mycoplasma pneumoniae infections. A major drawback of this technique is low specificity due to crossreactivity of LPS molecules in whole-cell antigen preparations. Greater specificity is possible with ELISA tests due to the specific antigen preparations (e.g. P1 adherence of Mycoplasma pneumoniae) that are used.