Anti-Pseudomonas aeruginosa ELISA Kit

Sample

Serum and plasma

Species Reactivity

Human

Detection Method

iELISA

Intended Use

The Anti-Pseudomonas aeruginosa ELISA Kit is an enzyme immunoassay for qualitative and quantitative detection of IgG-Antibodies in human serum against the extracellular proteins: Alkaline Protease (AP), Elastase (ELA) and Exotoxin A (Exo A) of Pseudomonas aeruginosa.

Contents of Kit

1. Microtiter plate 1 (AP): ready for use, coated with alkaline Protease, labelled red. Wells are separately breakable. (8x12) wells
2. Microtiter plate 2 (ELA): ready for use, coated with Elastase, labelled blue. Wells are separately breakable. (8x12) wells
3. Microtiter plate 3 (Exo A): ready for use, coated with Exotoxin A, labelled green. Wells are separately breakable. (8x12) wells
4. Positive Control 1 (PC1):, ready for use, calibrated control serum for alkaline Protease, AP, red labelled, shows a titer of 1:2500. 1 x 1.5 mL
5. Positive Control 2 (PC2):, ready for use, calibrated control serum for Elastase, ELA, blue labelled, shows a titer of 1:2500. 1 x 1.5 mL
6. Positive Control 3 (PC3):, ready for use, calibrated control serum for Exotoxin A, Exo A, green labelled, shows a titer of 1:2500. 1 x 1.5 mL
7. Negative Control (NC): ready for use, contains human serum, not reactive for Pseudomonas aeruginosa antigens. 2 x 1.5 mL
8. Control (CTR): ready for use, contains human serum and has to be determinated as borderline for Pseudomonas aeruginosa antigens in the assay 2 x 1.5 mL
9. Antibody-HRP-Conjugate (DET): 100-fold concentrated, contains horseradish peroxidase (HRP)-labelled anti-human IgG. 2 x 250 μL
10. Dilution Buffer (DIL): ready for use. Please shake before use! 3 x 50 mL
11. Washing Buffer (WB): 20-fold concentrated solution. 1 x 120 mL
12. Substrate (S): ready for use, horseradish-peroxidase-(HRP) substrate, stabilised Tetramethylbencidine. 1 x 33 mL
13. Stopping Solution (STP): ready for use, 0.2 M sulfuric acid. 1 x 33 mL
14. Sealing Tape: for covering the microtiter plate. 6

Storage

Store the kit at 2-8°C after receipt until its expiry date.
The shelf life of the components after initial opening is warranted for 4 weeks, store the unused strips and microtiter wells airtight together with the desiccant at 2-8°C in the clip-lock bag, use in the frame provided.

Precision

Intra-Assay Variance
Serum samples were diluted 1:1000 and measured 16-fold within one assay. Mean variance was 4.95, 4.49 and 8.41% for alkaline Protease, Elastase and Exotoxin A, respectively.

Inter-Assay Variance
Samples were portioned and stored at -20°C for up to 12 years and measured irregularly during this time. The variability measured as coefficient of variation does not exceed 20% for any antigen or sample.

Sensitivity

The analytical sensitivity as a measure for the minimal amount of specific antibody detectable by this test system was determined by the signal variability of the negative control. Therefore, the recalculated antibody titre of the negative control was evaluated and the 3-fold standard deviation was used to determine the limit of detection. The results are shown in table below. The limit of detection of antibody titers for AP, ELA, Exo A in 1:1000 diluted samples is < 1:300.

General Description

Pseudomonas aeruginosa, a Gram-negative bacterium ubiquitously distributed in the moist environment, causes about 10% of all nosocomial infections. This opportunistic pathogen leads to acute and chronic types of infection within various organs of susceptible patient groups. Chronic pulmonary infection in patients with cystic fibrosis (CF) is very frequent, may start early in infancy and determines the life expectancy of these patients. P. aeruginosa infection provokes a production of antibodies to a large number of P. aeruginosa antigens in CF patients. Creative Diagnostics's sensitive antibody detection system is an aid to discriminate between infected and uninfected patient groups.
Depending on the Pseudomonas aeruginosa species and the patient's immune reaction, antibodies can be detected against a single, two or even all three antigens simultaneously. A patient is regarded as sero-positive when the serum is positive for one or more of the antigens.
Used for research purposes and cannot be used as a diagnostic basis.